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Hair Cortisol, Perceived Stress and Dispositional Optimism:
A pilot study among Adolescents
Joel Milam PhD1, Rhona Slaughter MA, Gaurav Verma MA, & Rob McConnell MD
Department of Preventive Medicine
Keck School of Medicine
University of Southern California






1 Contact author:
Department of Preventive Medicine,		
Keck School of Medicine
University of Southern California,	
2001 Soto Building, SSB 3rd Floor, MC 9239
Los Angeles, CA  90032
Main Line: (323)442-8200
FAX: (323)442-8201
E-mail: HYPERLINK "mailto:milam@usc.edu"milam@usc.edu

Abstract
Background/Aims: Psychological stress is often associated with poor health-related outcomes.  One potential biomarker for chronic stress, hair cortisol, is minimally invasive compared to other cortisol collection techniques.  This pilot study examined the relationships between hair cortisol and self-reported perceived stress, stressful life events, depressive symptoms, and dispositional optimism among adolescents.
Methods: This cross-sectional study comprised of a convenience sample of 27 adolescents (age: M=14.96, SD=1.63) recruited from a Southern California after-school program.  Along with demographic and hair characteristics (e.g., hair color, type, etc.), participants completed the Perceived Stress Scale, Stressful Life Events checklist, CES-D (depressive symptoms), and Life Orientation Test (optimism). Hair cortisol was measured by analyzing hair samples approximately 1 cm from the scalp representing one month of cortisol exposure.
Results: Hair cortisol had a significant inverse association with dispositional optimism (r=-.44, p<.05).  Hair cortisol was not significantly associated with self-reported perceived stress, stressful life events, or depressive symptoms.
Conclusion: Assessment of hair cortisol may prove beneficial as an objective measure in research examining chronic stress-related outcomes among adolescents. Resiliency or protective dispositions, such as optimism, merit attention in relation to this biomarker.
Keywords: Hair cortisol, Stress, Stressful Life Events, Dispositional Optimism, Adolescents
Introduction
Psychological stress is a common risk factor for poor mental, behavioral, and physical health. Stress leads to the release of various pituitary and adrenal hormones that influence the immune system  ADDIN EN.CITE <EndNote><Cite><Author>Sapolsky</Author><Year>1994</Year><RecNum>877</RecNum><DisplayText>[1]</DisplayText><record><rec-number>877</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">877</key></foreign-keys><ref-type name="Book">6</ref-type><contributors><authors><author>Sapolsky, Robert, M.</author></authors></contributors><titles><title>Why zebras don&apos;t get ulcers : a guide to stress, stress related diseases, and coping</title></titles><pages>xii, 368 p.</pages><keywords><keyword>Stress (Physiology)</keyword><keyword>Stress (Psychology)</keyword><keyword>Stress management.</keyword></keywords><dates><year>1994</year></dates><pub-location>New York</pub-location><publisher>W.H. Freeman</publisher><isbn>0716723913</isbn><accession-num>635196</accession-num><call-num>Jefferson or Adams Building Reading Rooms - STORED OFFSITE QP82.2.S8; S266 1994</call-num><urls></urls></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_1" \o "Sapolsky, 1994 #877" 1].  Over time, chronic stress can lead to a dysregulated neuroendocrine or neuroimmune response, including a hypersensitivity to stressors marked by elevated levels of circulating cortisol and slow recoveries to basal levels. 
An important limitation to the field of stress research is that there has been no objective biomarker for chronic (vs. acute) stress.  Although repeated and pooled measures of stress biomarkers (e.g., saliva/urinary cortisol levels) have been used to assess chronic stress, these approaches are expensive and are logistically burdensome within the context of field studies of large populations. Further, the interpretation of the pattern of diurnal variation in salivary cortisol is also not well validated as an indicator of chronic stress, partially because of high levels of variance reflecting multiple diurnal patterns  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_2" \o "Fries, 2009 #879" 2]. 
	Hair cortisol is a promising biomarker for assessing chronic stress  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_3" \o "Gow, 2010 #853" 3].  Whereas salivary and plasma cortisol are indicators of acute stress (over hours), and urine collected appropriately may be an indicator for stress during the prior day, hair cortisol is an indicator of exposure during the period of growth of hair and therefore fills an important methodological gap  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_4" \o "Davenport, 2006 #844" 4,  HYPERLINK  \l "_ENREF_5" \o "Kirschbaum, 2009 #887" 5]. Hair grows approximately 1cm per month and the laboratory methodologies for assessing this hormone in hair have recently been developed and validated.  In the amounts needed, hair can be collected quickly and is acceptable to participants in culturally diverse settings  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_6" \o "Kim, 2009 #889" 6]. 
	Hair cortisol levels have been positively associated with self-reported levels of perceived stress among adults  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_7" \o "Kalra, 2007 #857" 7] and children  ADDIN EN.CITE <EndNote><Cite><Author>Vanaelst</Author><Year>2013</Year><RecNum>54</RecNum><DisplayText>[8]</DisplayText><record><rec-number>54</rec-number><foreign-keys><key app="EN" db-id="zpvd2d52srd5axeetv1psfz9zeaawrdrf5at">54</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Vanaelst, Barbara</author><author>Michels, Nathalie</author><author>Vriendt, Tineke</author><author>Huybrechts, Inge</author><author>Vyncke, Krishna</author><author>Sioen, Isabelle</author><author>Bammann, Karin</author><author>Rivet, Noellie</author><author>Raul, Jean-Sebastien</author><author>Molnar, Denes</author><author>Henauw, Stefaan</author></authors></contributors><titles><title>Cortisone in hair of elementary school girls and its relationship with childhood stress</title><secondary-title>European Journal of Pediatrics</secondary-title><alt-title>Eur J Pediatr</alt-title></titles><periodical><full-title>European Journal of Pediatrics</full-title><abbr-1>Eur J Pediatr</abbr-1></periodical><alt-periodical><full-title>European Journal of Pediatrics</full-title><abbr-1>Eur J Pediatr</abbr-1></alt-periodical><pages>843-846</pages><volume>172</volume><number>6</number><keywords><keyword>Child</keyword><keyword>Stress</keyword><keyword>Hair</keyword><keyword>Cortisone</keyword><keyword>Life events</keyword></keywords><dates><year>2013</year><pub-dates><date>2013/06/01</date></pub-dates></dates><publisher>Springer-Verlag</publisher><isbn>0340-6199</isbn><urls><related-urls><url>http://dx.doi.org/10.1007/s00431-013-1955-1</url></related-urls></urls><electronic-resource-num>10.1007/s00431-013-1955-1</electronic-resource-num><language>English</language></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_8" \o "Vanaelst, 2013 #54" 8] and is responsive to specific major life stressors.  For example, among rhesus monkeys, hair cortisol levels increased following moving them to a new location  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_4" \o "Davenport, 2006 #844" 4].  Among neonates, days on a ventilator (a chronic stressor) was associated with elevated hair cortisol levels  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_9" \o "Yamada, 2007 #875" 9].  Among young children, hair cortisol levels increased following school entry ADDIN EN.CITE <EndNote><Cite><Author>Groeneveld</Author><Year>2013</Year><RecNum>55</RecNum><DisplayText>[10]</DisplayText><record><rec-number>55</rec-number><foreign-keys><key app="EN" db-id="zpvd2d52srd5axeetv1psfz9zeaawrdrf5at">55</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Marleen G. Groeneveld</author><author>Harriet J. Vermeer</author><author>Mari�lle Linting</author><author>Gerard Noppe</author><author>Elisabeth F. C. van Rossum</author><author>Marinus H. van IJzendoorn</author></authors></contributors><titles><title>Children�s hair cortisol as a biomarker of stress at school entry</title><secondary-title>Stress</secondary-title></titles><periodical><full-title>Stress</full-title></periodical><pages>1-5</pages><volume>0</volume><number>0</number><dates><year>2013</year></dates><accession-num>23786528</accession-num><urls><related-urls><url>http://informahealthcare.com/doi/abs/10.3109/10253890.2013.817553</url></related-urls></urls><electronic-resource-num>doi:10.3109/10253890.2013.817553</electronic-resource-num></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_10" \o "Groeneveld, 2013 #55" 10]; and among adults, those with long term unemployment (12+ months) had higher hair cortisol levels than those currently employed  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_11" \o "Dettenborn, 2010 #845" 11].  Moreover, adults with chronic pain had higher hair cortisol levels than those without chronic pain  ADDIN EN.CITE <EndNote><Cite><Author>Van Uum</Author><Year>2008</Year><RecNum>873</RecNum><DisplayText>[12]</DisplayText><record><rec-number>873</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">873</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Van Uum, S. H.</author><author>Sauve, B.</author><author>Fraser, L. A.</author><author>Morley-Forster, P.</author><author>Paul, T. L.</author><author>Koren, G.</author></authors></contributors><auth-address>Division of Endocrinology and Metabolism, University of Western Ontario, Ontario, Canada. Stan.VanUum@sjhc.london.on.ca</auth-address><titles><title>Elevated content of cortisol in hair of patients with severe chronic pain: a novel biomarker for stress</title><secondary-title>Stress</secondary-title><alt-title>Stress</alt-title></titles><periodical><full-title>Stress</full-title><abbr-1>Stress</abbr-1></periodical><alt-periodical><full-title>Stress</full-title><abbr-1>Stress</abbr-1></alt-periodical><pages>483-8</pages><volume>11</volume><number>6</number><edition>2008/07/09</edition><keywords><keyword>Adult</keyword><keyword>Analgesics, Opioid/therapeutic use</keyword><keyword>Biological Markers/*analysis</keyword><keyword>Chronic Disease</keyword><keyword>Female</keyword><keyword>Hair/*chemistry</keyword><keyword>Humans</keyword><keyword>Hydrocortisone/*analysis</keyword><keyword>Male</keyword><keyword>Middle Aged</keyword><keyword>Pain/*diagnosis/drug therapy</keyword><keyword>Stress, Psychological/*metabolism</keyword></keywords><dates><year>2008</year></dates><isbn>1607-8888 (Electronic)&#xD;1025-3890 (Linking)</isbn><accession-num>18609301</accession-num><work-type>Research Support, Non-U.S. Gov&apos;t</work-type><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/pubmed/18609301</url></related-urls></urls><electronic-resource-num>10.1080/10253890801887388</electronic-resource-num><language>eng</language></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_12" \o "Van Uum, 2008 #873" 12].  However, other studies among adults found no association between the hair cortisol and PSS, depressive symptoms, or chronic stress  ADDIN EN.CITE  ADDIN EN.CITE.DATA [e.g.,  HYPERLINK  \l "_ENREF_13" \o "Dowlati, 2010 #848" 13,  HYPERLINK  \l "_ENREF_12" \o "Van Uum, 2008 #873" 12,  HYPERLINK  \l "_ENREF_11" \o "Dettenborn, 2010 #845" 11] and one study among young adults found an inverse association between hair cortisol and perceived stress and depressive symptoms  ADDIN EN.CITE <EndNote><Cite><Author>Gerber</Author><Year>2013</Year><RecNum>56</RecNum><DisplayText>[14]</DisplayText><record><rec-number>56</rec-number><foreign-keys><key app="EN" db-id="zpvd2d52srd5axeetv1psfz9zeaawrdrf5at">56</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Gerber, M.</author><author>Kalak, N.</author><author>Elliot, C.</author><author>Holsboer-Trachsler, E.</author><author>P�hse, U.</author><author>Brand, S.</author></authors></contributors><titles><title>Both Hair Cortisol Levels and Perceived Stress Predict Increased Symptoms of Depression: An Exploratory Study in Young Adults</title><secondary-title>Neuropsychobiology</secondary-title></titles><periodical><full-title>Neuropsychobiology</full-title></periodical><pages>100-109</pages><volume>68</volume><number>2</number><dates><year>2013</year></dates><isbn>0302-282X</isbn><urls><related-urls><url>http://www.karger.com/DOI/10.1159/000351735</url></related-urls></urls></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_14" \o "Gerber, 2013 #56" 14].  Thus, the relationship between hair cortisol and self-reported stress may depend on the characteristics of the study population.    
	No studies have examined the relationship between hair cortisol and stress-buffering/protective factors.  Dispositions that mitigate the impact of life stressors may have a stronger relationship with biomarkers of chronic stress (i.e., beyond 1 month) than shorter term indicators of stress because these dispositions are less resistant to change and may have greater cumulative effects over time.  Dispositional optimism has well-established protective relationships with both perceived stress and stressful life events (e.g., through greater use of adaptive strategies for coping with stressors) ADDIN EN.CITE <EndNote><Cite><Author>Carver</Author><Year>2010</Year><RecNum>891</RecNum><DisplayText>[15]</DisplayText><record><rec-number>891</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">891</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Carver, C. S.</author><author>Scheier, M. F.</author><author>Segerstrom, S. C.</author></authors></contributors><auth-address>Department of Psychology, University of Miami, Coral Gables, FL 33124-0751, USA. Ccarver@miami.edu</auth-address><titles><title>Optimism</title><secondary-title>Clinical Psychology Review</secondary-title><alt-title>Clin Psychol Rev</alt-title></titles><periodical><full-title>Clinical Psychology Review</full-title></periodical><pages>879-89</pages><volume>30</volume><number>7</number><edition>2010/02/23</edition><keywords><keyword>*Adaptation, Psychological</keyword><keyword>*Emotions</keyword><keyword>Health Behavior</keyword><keyword>Humans</keyword><keyword>Personal Satisfaction</keyword><keyword>*Personality</keyword></keywords><dates><year>2010</year><pub-dates><date>Nov</date></pub-dates></dates><isbn>1873-7811 (Electronic)&#xD;0272-7358 (Linking)</isbn><accession-num>20170998</accession-num><work-type>Research Support, N.I.H., Extramural&#xD;Research Support, U.S. Gov&apos;t, Non-P.H.S.&#xD;Review</work-type><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/pubmed/20170998</url></related-urls></urls><electronic-resource-num>10.1016/j.cpr.2010.01.006</electronic-resource-num><language>eng</language></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_15" \o "Carver, 2010 #891" 15].  In addition, stress is one potential pathway through which optimism positively influences various physical health outcomes  ADDIN EN.CITE <EndNote><Cite><Author>Rasmussen</Author><Year>2009</Year><RecNum>905</RecNum><DisplayText>[16]</DisplayText><record><rec-number>905</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">905</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Rasmussen, H. N.</author><author>Scheier, M. F.</author><author>Greenhouse, J. B.</author></authors></contributors><auth-address>Institute for Educational Research and Public Service, University of Kansas, Lawrence, KS 66045, USA.</auth-address><titles><title>Optimism and physical health: a meta-analytic review</title><secondary-title>Annals of behavioral medicine : a publication of the Society of Behavioral Medicine</secondary-title><alt-title>Ann Behav Med</alt-title></titles><periodical><full-title>Annals of behavioral medicine : a publication of the Society of Behavioral Medicine</full-title><abbr-1>Ann Behav Med</abbr-1></periodical><alt-periodical><full-title>Annals of behavioral medicine : a publication of the Society of Behavioral Medicine</full-title><abbr-1>Ann Behav Med</abbr-1></alt-periodical><pages>239-56</pages><volume>37</volume><number>3</number><edition>2009/08/28</edition><keywords><keyword>*Attitude</keyword><keyword>*Health Status</keyword><keyword>*Health Status Indicators</keyword><keyword>Humans</keyword></keywords><dates><year>2009</year><pub-dates><date>Jun</date></pub-dates></dates><isbn>1532-4796 (Electronic)&#xD;0883-6612 (Linking)</isbn><accession-num>19711142</accession-num><work-type>Meta-Analysis&#xD;Research Support, N.I.H., Extramural&#xD;Review</work-type><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/pubmed/19711142</url></related-urls></urls><custom2>2941870</custom2><electronic-resource-num>10.1007/s12160-009-9111-x</electronic-resource-num><language>eng</language></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_16" \o "Rasmussen, 2009 #905" 16].  For example, dispositional optimism is associated with lower salivary cortisol awakening response  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_17" \o "Endrighi, 2011 #2041" 17,  HYPERLINK  \l "_ENREF_18" \o "Lai, 2005 #2042" 18].  Thus, dispositional optimism is an important contributor to consider in relation to biomarkers of chronic stress.  
	The primary aim of the current study was to examine the relationships between hair cortisol and self-reported perceived stress, stressful life events, depressive symptoms, and dispositional optimism among adolescents.  We hypothesized that hair cortisol would be positively associated with perceived stress and stressful life events scales and inversely associated with dispositional optimism.
Method
Participants.  Participants included a convenience sample of 28, primarily Hispanic (93%), adolescents (mean age = 14.96, years, SD = 1.63, 70% female) recruited from a local after school program.  One participant was excluded due to unrealistic survey responses (i.e., responded positively to all 65 stressful life events).  Personal assent and parental informed consent were obtained from all participants, and hair samples and self-report surveys were collected using a protocol approved by Institutional Review Board of the University of Southern California.  The total procedure required approximately 20 minutes and participants received a $10 gift card for their time. 
Measures
Demographics and hair characteristics. We assessed age, gender, hair color, hair type, hair treatment/dying, hair blow drying, and use of hand creams (which may contain hydrocortisone).
Perceived Stress. The 10 item Perceived Stress Scale was used to assess stress over the prior month (PSS)  ADDIN EN.CITE <EndNote><Cite><Author>Cohen</Author><Year>1988</Year><RecNum>1529</RecNum><DisplayText>[19]</DisplayText><record><rec-number>1529</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">1529</key></foreign-keys><ref-type name="Book Section">5</ref-type><contributors><authors><author>Cohen, S.</author><author>Williamson G.</author></authors><secondary-authors><author>Spacapan, S.</author><author>Oskamp S.</author></secondary-authors></contributors><titles><title>Perceived stress in a probability sample of the United States</title><secondary-title>The Social Psychology of Health</secondary-title></titles><pages>31-67</pages><dates><year>1988</year></dates><pub-location>Newbury Park, CA</pub-location><publisher>Sage</publisher><urls></urls></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_19" \o "Cohen, 1988 #1529" 19]. The PSS assesses the extent respondents felt their lives were unpredictable, uncontrollable, or overwhelming over the prior month (e.g., �In the last month, how often have you felt that you were unable to control the important things in your life?).  Each item has a 5-point response format ranging from �never� to �very often.�  
Stressful Life Events. We used a well-validated and age appropriate 65-item check list of stressful life events experienced over the prior 3 months  ADDIN EN.CITE <EndNote><Cite><Author>Booker</Author><Year>2004</Year><RecNum>1612</RecNum><DisplayText>[20]</DisplayText><record><rec-number>1612</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">1612</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Booker, C.</author><author>Gallaher, P.</author><author>Unger, J.</author><author>Ritt-Olson, A.</author><author>Johnson, C. A.</author></authors></contributors><titles><title>Stressful life events, smoking behavior, and intentions to smoke among a multiethnic sample of sixth graders.  </title><secondary-title>Ethnicity &amp; Health</secondary-title></titles><periodical><full-title>Ethnicity &amp; Health</full-title></periodical><pages>369-397</pages><volume>9</volume><number>4</number><dates><year>2004</year></dates><urls></urls></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_20" \o "Booker, 2004 #1612" 20].  This scale was developed and validated among a sample of 1,074 multi-ethnic youth (mean age = 12.75) from urban Los Angeles middle schools and includes a range of commonly experienced events spanning from temporary (e.g., test taking, arguments with friends) to more severe (e.g., injury, family death) stressors.  For each endorsed event, the stressfulness of the event was rated on a 1 (not at all stressful) to 4 (very stressful) scale.  The overall sum of endorsed events was used to form a total stressful life event score for each participant.  A weighted sum was used to form a total life event stressfulness score for each participant.
Depressive symptoms was ascertained with the widely used Center for Epidemiological Studies Depression Scale (CES-D)  ADDIN EN.CITE <EndNote><Cite><Author>Radloff</Author><Year>1977</Year><RecNum>376</RecNum><DisplayText>[21]</DisplayText><record><rec-number>376</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">376</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Radloff, L. S.</author></authors></contributors><titles><title>The CES-D scale: A self-report depression scale for research in the general population</title><secondary-title>Applied Psychological Measurement</secondary-title></titles><periodical><full-title>Applied Psychological Measurement</full-title></periodical><pages>385-401</pages><volume>1</volume><number>3</number><dates><year>1977</year></dates><urls></urls></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_21" \o "Radloff, 1977 #376" 21].  The CES-D is a 20-item measure of current (i.e., past week) depressive symptoms (e.g., �I felt depressed,� �I felt lonely,� and �I felt that everything I did was an effort�).  Responses are made on a 4-point scale, ranging from 0 (rarely or none of the time, less than one day) to 3 (most or all of the time, 5-7 days).  
Optimism was assessed with the revised Life Orientation Test (LOT-R)  ADDIN EN.CITE <EndNote><Cite><Author>Scheier</Author><Year>1994</Year><RecNum>23</RecNum><DisplayText>[22]</DisplayText><record><rec-number>23</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">23</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Scheier, M. F.</author><author>Carver, C. S.</author><author>Bridges, M. W.</author></authors></contributors><titles><title>Distinguishing optimism from neuroticism (and trait anxiety, self-mastery, and self-esteem): a reevaluation of the Life Orientation Test</title><secondary-title>Journal of Personality &amp; Social Psychology</secondary-title></titles><periodical><full-title>Journal of Personality &amp; Social Psychology</full-title></periodical><pages>1063-78</pages><volume>67</volume><number>6</number><keywords><keyword>Adaptation, Psychological</keyword><keyword>*Anxiety</keyword><keyword>Comparative Study</keyword><keyword>Female</keyword><keyword>Human</keyword><keyword>Male</keyword><keyword>Self Assessment (Psychology)</keyword><keyword>*Self Concept</keyword><keyword>Support, Non-U.S. Gov&apos;t</keyword><keyword>Support, U.S. Gov&apos;t, Non-P.H.S.</keyword><keyword>Support, U.S. Gov&apos;t, P.H.S.</keyword></keywords><dates><year>1994</year></dates><urls></urls></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_22" \o "Scheier, 1994 #23" 22]. This widely used 6 item scale assesses future life expectancies using a 5-point response format ranging from �strongly agree� to �strongly disagree.�  Examples of items include �In uncertain times I usually expect the best� and �I rarely count on good things happening to me� (reverse scored).  
Hair samples. At least 10mg (~50 strands of hair) was collected from the vertex posterior, where there is the most uniform growth pattern between individuals, using procedures previously developed  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_4" \o "Davenport, 2006 #844" 4].  Fine tipped surgical scissors were used to obtain hair strands as close to the scalp as possible.  Hair samples were placed in foil, indicated with the direction of the scalp end, labeled with participant ID, and placed in a plastic bag for transportation and storage.  
Hair cortisol assessment. Between 10-20 mg of hair from the 1cm closest to the scalp end was utilized to assess cortisol using a protocol developed by Davenport et al. (2006) ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_4" \o "Davenport, 2006 #844" 4].  Each hair segment is placed into a falcon tube, isopropanol is added, and the tube mixed using an overhead rotator for 3 minutes.  After decanting, the wash cycle is repeated and the sample is dried, powdered, and weighed (50 mg) into a cryo vial.  Methanol is added (with vials rotated for 24 h) for extraction of the steroid.  Samples are spun (microcentrifuge at 10 rpm for 2 min) and 1 ml of the clear supernatant is transferred into a new cryo vial.  The alcohol is evaporated and twenty microliters are used for cortisol determination using a commercially available immunoassay with chemiluminescence detection (CLIA, IBL-Hamburg, Germany).  Intra- and interassay coefficient of variance of this assay is below 8% and  validation work (among adults) using liquid chromatography mass spectrometry/MS has shown a strong correlation (r-square = .96); CLIA results were linearly inflated over the LCMS/MS results by 15%, an acceptable deviation  ADDIN EN.CITE <EndNote><Cite><Author>Kirschbaum</Author><Year>2009</Year><RecNum>887</RecNum><DisplayText>[5]</DisplayText><record><rec-number>887</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">887</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Kirschbaum, C.</author><author>Tietze, A.</author><author>Skoluda, N.</author><author>Dettenborn, L.</author></authors></contributors><auth-address>Department of Psychology, Technical University of Dresden, 01062 Dresden, Germany. ck@biopsych.tu-dresden.de</auth-address><titles><title>Hair as a retrospective calendar of cortisol production-Increased cortisol incorporation into hair in the third trimester of pregnancy</title><secondary-title>Psychoneuroendocrinology</secondary-title><alt-title>Psychoneuroendocrinology</alt-title></titles><periodical><full-title>Psychoneuroendocrinology</full-title></periodical><alt-periodical><full-title>Psychoneuroendocrinology</full-title></alt-periodical><pages>32-7</pages><volume>34</volume><number>1</number><edition>2008/10/25</edition><keywords><keyword>Female</keyword><keyword>Hair/*chemistry/metabolism</keyword><keyword>Humans</keyword><keyword>Hydrocortisone/*analysis/metabolism</keyword><keyword>Postpartum Period</keyword><keyword>Pregnancy</keyword><keyword>*Pregnancy Trimester, Third</keyword><keyword>Retrospective Studies</keyword><keyword>Time Factors</keyword></keywords><dates><year>2009</year><pub-dates><date>Jan</date></pub-dates></dates><isbn>0306-4530 (Print)&#xD;0306-4530 (Linking)</isbn><accession-num>18947933</accession-num><work-type>Evaluation Studies&#xD;Research Support, Non-U.S. Gov&apos;t</work-type><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/pubmed/18947933</url></related-urls></urls><electronic-resource-num>10.1016/j.psyneuen.2008.08.024</electronic-resource-num><language>eng</language></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_5" \o "Kirschbaum, 2009 #887" 5].  Due to skewness, hair cortisol levels were log transformed for all analyses and exponentiated for presentation.  We present results for the first segment of hair to retain the largest sample size.
Results
Independent sample T-test comparisons of cortisol levels did not differ between participants who: had treated (bleached/dyed/colored) their hair in the past 3 months (30.8%) vs. not (69.2%), had their hair permed or straightened in the past 3 months (48.1%) vs. not (51.9%), had black (51.9%) vs. brown (40.7%) hair, or had straight (48.1%) vs. wavy/curly (51.9%) hair (all p�s>.05).  Hair cortisol was not associated with number of days per week hair was shampooed (all p values>.21).
Descriptive data and correlations of the main study variables are located in Table 1.  Mean levels of hair cortisol were similar to those previously reported in populations of different ages (e.g.,  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_23" \o "Steudte, 2011 #884" 23]. Hair cortisol levels were not significantly related to self-reported perceived stress, life event checklist summary scores, life event stressfulness scores, or reports of depressive symptoms. There was a large significant inverse relationship between hair cortisol and dispositional optimism.  Optimism also had a significant inverse relationship with perceived stress and depressive symptoms.
Hair cortisol was regressed against optimism scores in a multivariable model controlling for gender, perceived stress, and depressive symptoms.  The overall model approached significance (F(4,25) = 2.63, p=.06; R2=.33), with a significant association between hair cortisol and optimism (B=-.57, t=-3.07, p<.01).
Discussion
	These preliminary data support hair cortisol as a potential biomarker of chronic cortisol exposure among younger populations.  Dispositional optimism is linked with beneficial physical responses to stress e.g., ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_24" \o "Thomas, 2011 #1708" 24,  HYPERLINK  \l "_ENREF_25" \o "Terrill, 2010 #1717" 25], including lower levels of salivary cortisol awakening response  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_17" \o "Endrighi, 2011 #2041" 17,  HYPERLINK  \l "_ENREF_18" \o "Lai, 2005 #2042" 18], and to our knowledge this is the first study to find a similar relationship with a biomarker for chronic stress.  Thus, more stable trait-like measures may prove useful in future work examining hair cortisol.  For example, personality characteristics, such as neuroticism and hostility, and chronic stressful neighborhood characteristics, such as crime or noise  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_26" \o "Steptoe, 2001 #1996" 26], might have a greater effect on chronic cortisol exposure because they are more resistant to change than short term stressors.
There are numerous potential reasons that there were no significant relationships between hair cortisol and self-reported perceived stress or stressful life events.  First, our sample size was limited, reducing our statistical power, and there may have been unknown factors related to unique characteristics of the participants.  Nevertheless, the relatively small to medium relationships (r�s=.14-.32) are consistent with prior work  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_13" \o "Dowlati, 2010 #848" 13,  HYPERLINK  \l "_ENREF_12" \o "Van Uum, 2008 #873" 12] and suggests low to modest overlap between the self-reported stress measures and hair cortisol.    
Second, although our assessments of stress were well-validated and temporally aligned to the collected samples, they may not have adequately reflected ongoing chronic stress.  For example, the prior 3 month life event checklist (and perceived stress for the prior 30 days) may have captured events that influenced cortisol levels temporarily, but not long enough to impact levels detected in hair.  Repeated assessments of life events and both serum/urinary and hair cortisol levels (to reflect more acute and chronic cortisol exposure, respectively) may be able to determine whether chronic stressor exposure has a greater long term impact on HPA activity.  Prior work with chronic unemployment  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_11" \o "Dettenborn, 2010 #845" 11] and chronic pain  ADDIN EN.CITE <EndNote><Cite><Author>Van Uum</Author><Year>2008</Year><RecNum>873</RecNum><DisplayText>[12]</DisplayText><record><rec-number>873</rec-number><foreign-keys><key app="EN" db-id="ee200fe0n2d0flesxpbp9e2u2az2rrra5s9z">873</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Van Uum, S. H.</author><author>Sauve, B.</author><author>Fraser, L. A.</author><author>Morley-Forster, P.</author><author>Paul, T. L.</author><author>Koren, G.</author></authors></contributors><auth-address>Division of Endocrinology and Metabolism, University of Western Ontario, Ontario, Canada. Stan.VanUum@sjhc.london.on.ca</auth-address><titles><title>Elevated content of cortisol in hair of patients with severe chronic pain: a novel biomarker for stress</title><secondary-title>Stress</secondary-title><alt-title>Stress</alt-title></titles><periodical><full-title>Stress</full-title><abbr-1>Stress</abbr-1></periodical><alt-periodical><full-title>Stress</full-title><abbr-1>Stress</abbr-1></alt-periodical><pages>483-8</pages><volume>11</volume><number>6</number><edition>2008/07/09</edition><keywords><keyword>Adult</keyword><keyword>Analgesics, Opioid/therapeutic use</keyword><keyword>Biological Markers/*analysis</keyword><keyword>Chronic Disease</keyword><keyword>Female</keyword><keyword>Hair/*chemistry</keyword><keyword>Humans</keyword><keyword>Hydrocortisone/*analysis</keyword><keyword>Male</keyword><keyword>Middle Aged</keyword><keyword>Pain/*diagnosis/drug therapy</keyword><keyword>Stress, Psychological/*metabolism</keyword></keywords><dates><year>2008</year></dates><isbn>1607-8888 (Electronic)&#xD;1025-3890 (Linking)</isbn><accession-num>18609301</accession-num><work-type>Research Support, Non-U.S. Gov&apos;t</work-type><urls><related-urls><url>http://www.ncbi.nlm.nih.gov/pubmed/18609301</url></related-urls></urls><electronic-resource-num>10.1080/10253890801887388</electronic-resource-num><language>eng</language></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_12" \o "Van Uum, 2008 #873" 12] further support this possibility.  
Third, there may be a �lack of psychoendocrine covariance�  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_11" \o "Dettenborn, 2010 #845" e.g., 11].  That is, psychological stress is a dynamic process and people may report psychological stress when they are not stressed physiologically and alternatively may be physiologically stressed, but not report any psychological stress.  Because most research examining chronic stress utilizes self-report assessments, hair cortisol may provide a more reliable estimate of physiologic stress exposure.  Thus, this approach may be particularly helpful in delineating the relationships between chronic stress and inflammatory markers among people living with chronic illnesses, such as HIV or asthma.  
Hair cortisol is a potential indicator for assessing chronic stress exposure, particularly in larger studies among younger populations where response burden to in-depth interviews of psychological stress can be difficult.  Because hair cortisol is relatively inexpensive to obtain and transport, significantly less invasive than serum sampling, not reactive to/biased from the assessment method/procedure, and is more feasible than extensive standardized psychological surveys or interviews within the context of large cohort studies, it has great potential for use in research among diverse and urban populations.  This is potentially useful in studies of more vulnerable populations (e.g., low socioeconomic status, immunocompromised, children) where chronic stress is posited to disproportionately contribute towards the development of inflammatory chronic diseases  ADDIN EN.CITE  ADDIN EN.CITE.DATA [e.g.,  HYPERLINK  \l "_ENREF_27" \o "Taylor, 2004 #2005" 27,  HYPERLINK  \l "_ENREF_28" \o "Baum, 1999 #2017" 28-30] and increase the deleterious impact of physical risk factors for inflammatory outcomes  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_31" \o "Shankardass, 2009 #2026" e.g., air pollution; 31].  Thus, hair cortisol may serve as an important indicator to consider in research examining resiliency factors that are posited to buffer the impact of stress on various health outcomes  ADDIN EN.CITE  ADDIN EN.CITE.DATA [ HYPERLINK  \l "_ENREF_32" \o "Pressman, 2005 #52" 32]  ADDIN EN.CITE <EndNote><Cite><Author>Steptoe</Author><Year>2009</Year><RecNum>53</RecNum><DisplayText>[33]</DisplayText><record><rec-number>53</rec-number><foreign-keys><key app="EN" db-id="zpvd2d52srd5axeetv1psfz9zeaawrdrf5at">53</key></foreign-keys><ref-type name="Journal Article">17</ref-type><contributors><authors><author>Steptoe, Andrew</author><author>Dockray, Samantha</author><author>Wardle, Jane</author></authors></contributors><titles><title>Positive Affect and Psychobiological Processes Relevant to Health</title><secondary-title>Journal of Personality</secondary-title></titles><periodical><full-title>Journal of Personality</full-title></periodical><pages>1747-1776</pages><volume>77</volume><number>6</number><dates><year>2009</year></dates><publisher>Blackwell Publishing Inc</publisher><isbn>1467-6494</isbn><urls><related-urls><url>http://dx.doi.org/10.1111/j.1467-6494.2009.00599.x</url></related-urls></urls><electronic-resource-num>10.1111/j.1467-6494.2009.00599.x</electronic-resource-num></record></Cite></EndNote>[ HYPERLINK  \l "_ENREF_33" \o "Steptoe, 2009 #53" 33].

Acknowledgements:
We thank Dr. Clemens Kirschbaum and his lab for performing the hair cortisol assays.

NIH Funding:
Research reported in this publication was supported by the National Cancer Institute of the National Institutes of Health under award number T32CA009492.





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