Name: collecting hair samples for hair cortisol analysis in african americans
Uploaded: 2018-06-10T14:00:00
Description: Watch this Scientific Journal Video about Collecting Hair Samples for Hair Cortisol Analysis in African Americans at JoVE.com

This publication was made possible by the Clinical and Translational Science Collaborative of Cleveland; KL2TR000440 from the National Center for Advancing Translational Sciences (NCATS) component of the National Institutes of Health; and NIH Roadmap for Medical Research. Dr. Jodi Ford's work in this area has been funded by the National Institutes of Health National Institute on Drug Abuse (1R21DA034960) and The Ohio State University Institute for Population Research seed grant funding from the National Institute of Child Health and Development (2P2CHD058484). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Dr. Laudenslager's work in this area has been funded by a contract from the Patient-Centered Outcomes Research Institute (CE-1304-6208), a grant from the Administration for Children and Families (90YR0058), and a grant from the Developmental Psychobiology Research Group Endowment Fund of the Department of Psychiatry, University of Colorado Denver Anschutz Medical Campus.

The study is approved by the University Hospitals Cleveland Medical Center Institutional Review Board, and the protocol follows the guidelines of participant protection and data safety as indicated by the review committee.
1. Prepare Participant for Hair Sample and Hair Sample Collection Questions
<ol>
	<li>Explain to participant that the hair sample collected is less hair than that is lost normally in brushing each day from the back of the head (posterior vertex). Tell the participant that ...

<ol>
	<li>Sauve, B. K. G., Walsh, G., Tokmakejian, S., Van Uum, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+cortisol+in+human+hair+as+a+biomarker+of+systemic+exposure.">Measurement of cortisol in human hair as a biomarker of systemic exposure.</a> Clin Invest Med. 30 (5), 183-191 (2007).</li><li>Wright, K. D., Hickman, R., Laudenslager, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hair+cortisol+analysis:+a+promising+biomarker+of+HPA+activation+in+older+adults.">Hair cortisol analysis: a promising biomarker of HPA activation in older adults.</a> Gerontologist. 55, 140-145 (2015).</li><li>Awad, G. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Beauty+and+body+image+concerns+among+African+American+college+women.">Beauty and body image concerns among African American college women.</a> J Black Psychol. 41 (6), 540-564 (2015).</li><li>Capodilupo, C. M., Kim, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gender+and+race+matter:+the+importance+of+considering+intersections+in+Black+women's+body+image.">Gender and race matter: the importance of considering intersections in Black women's body image.</a> J Couns Psychol. 61 (1), 37-49 (2014).</li><li>Miracle, V. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Belmont+Report:+the+triple+crown+of+research+ethics.">The Belmont Report: the triple crown of research ethics.</a> Dimens Crit Care Nurs. 35 (4), 223-228 (2016).</li><li>Fishman, B. M., Bobo, L., Kosub, K., Womeodu, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cultural+issues+in+serving+minority+populations:+emphasis+on+Mexican+Americans+and+African+Americans.">Cultural issues in serving minority populations: emphasis on Mexican Americans and African Americans.</a> Am J Med Sci. 306 (3), 160-166 (1993).</li><li>Webb, J. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Louisiana+voodoo+and+superstitions+related+to+health.">Louisiana voodoo and superstitions related to health.</a> HSMHA Health Rep. 86 (4), 291-301 (1971).</li><li>Ford, J. L., Boch, S. J., McCarthy, D. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Feasibility+of+hair+collection+for+cortisol+measurement+in+population+research+on+adolescent+health.">Feasibility of hair collection for cortisol measurement in population research on adolescent health.</a> Nurs Res. 65 (3), 249-255 (2016).</li><li>Wosu, A. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hair+cortisol+in+relation+to+sociodemographic+and+lifestyle+characteristics+in+a+multiethnic+US+sample.">Hair cortisol in relation to sociodemographic and lifestyle characteristics in a multiethnic US sample.</a> Ann Epidemiol. 25 (2), 90-95 (2015).</li><li>Webb, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+individual+systemic+stress+through+cortisol+analysis+of+archaeological+hair.">Assessing individual systemic stress through cortisol analysis of archaeological hair.</a> J Archaeolog Sci. 37 (4), 807-812 (2010).</li><li>Russell, E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+standardization+of+hair+cortisol+measurement:+results+of+the+first+international+interlaboratory+round+robin.">Toward standardization of hair cortisol measurement: results of the first international interlaboratory round robin.</a> Ther Drug Monit. 37 (1), 71-75 (2015).</li><li>Bosley, R. E., Daveluy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+primer+to+natural+hair+care+practices+in+black+patients.">A primer to natural hair care practices in black patients.</a> Cutis. 95 (2), 78-80 (2015).</li><li>Loussouarn, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=African+hair+growth+parameters.">African hair growth parameters.</a> Brit J Derm. 145 (2), 294-297 (2001).</li><li>Hoffman, M. C., Karban, L. V., Goodteacher, A., Laudenslager, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemical+processing+and+shampooing+impact+cortisol+measured+in+human+hair.">Chemical processing and shampooing impact cortisol measured in human hair.</a> Clin Invest Med. 37 (4), 252-257 (2014).</li><li>Ramirez, J., Elmofty, M., Castillo, E., DeRouen, M., Shariff-Marco, S., Allen, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+cortisol+and+telomere+length+measurements+in+ethnically+diverse+women+with+breast+cancer+using+culturally+sensitive+methods.">Evaluation of cortisol and telomere length measurements in ethnically diverse women with breast cancer using culturally sensitive methods.</a> J Community Genet. 8, 75-86 (2017).</li></ol>

Essential steps in the hair collection protocol include providing an example of how much hair will be taken, showing the sample site (posterior vortex), use of the single strain twist and gently pull method, and securing the sample with painter&#39;s tape away from the scalp end. By gently twisting and pulling the sample, the curly hair is lengthened and the scalp end alignment is maintained. This reduces laboratory time needed to align the scalp ends, measure, and cut the sample. Although thinning shears have been used ...

Collecting samples of saliva, blood, or urine for cortisol analysis has unique procedural challenges and greater participant burden, as the procedure requires multiple saliva samples, venipuncture samples, or 24-hour urine collection. Obtaining cortisol from hair samples is a simple and acceptable alternative to using saliva, serum, or urine samples1,2. Hair cortisol provides a retrospective representation of hypothalamic-pituitary-adrenal (HPA) axis function over time. Approximately 3 cm of hair collected from the scalp is equal to average cortisol release for the past 3 months1. However, researchers encounter unique challenges when attempting to collect hair samples across diverse populations. Such challenges include religious or spiritual beliefs including exposing hair (e.g., Muslim women), hair as sacred (American Indian groups), or cultural norms that connect hair to standards of beauty and self-image.
In the African American culture, particularly among women, scalp hair is a very personal and important aspect of their body image3,4. Further, historical experiences (e.g., the Tuskegee experiments) continue to foster mistrust in the research process5. Some African American folklore and cultural beliefs also suggest that the hair sample could be used in rituals that could cause harm6,7. Researchers should be aware of potential challenges that they may face in studies when African American participants are asked to provide hair samples.
In addition to issues surrounding body image, the specific techniques for collecting and handling hair samples from African Americans may differ from that of European Americans6,7. For example, African Americans may have straight, thin, curly, kinky, coarse, or a combination of these hair types, each requiring different techniques to collect and secure adequate hair samples for processing. Recent studies on samples of African American teens8, as well as multi-ethnic samples9, showed that collection of hair for cortisol level measurement was acceptable among African American participants. At present, there are no visual training procedures designed to provide researchers with the skill and cultural knowledge to collect hair samples in African Americans. The purpose of this paper is to present a culturally informed protocol to help researchers obtain adequate hair samples from African American participants. This method was developed with careful consideration of physical, psychosocial, and cultural characteristics relevant to collection of hair samples from African American adults. To date, there are no visual methods of collecting these samples, and this protocol will be the first guide that details proper hair sample collection from African American adults. To illustrate the representative results of the protocol implementation, de-identified data are presented from African Americans that participated in a community-based study on chronic stress that were aged 65 and older. Both male and female research assistants from a variety of racial backgrounds contacted potential participants from a clinic generated list and flyers. The primary investigator, who is an African American woman, spoke at community centers and senior subsidized housing apartments to older adult African Americans about the study, including a description of the hair collection methods.

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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">&#160;Salon grade scissors</td>
			<td style="width:232px;">Cricket</td>
			<td style="width:312px;">S-3 550</td>
			<td style="width:479px;">Carded Professional Hair Shear, 5.5 inch</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Large tooth comb</td>
			<td style="width:232px;">Goody</td>
			<td style="width:312px;">No catalog number__Goody Stylista Rake Comb&#160;&#160;&#160;</td>
			<td>Link to Company website&#160; http://www.goody.com/Products/Styling_Tools/Goody_Stylista_Rake_Comb?colId=d11c04a1-1dd3-4848-a313-046fc6c109ec&#38;c_id=35a36c88-d359-4104-a53c-dfbfd8452780</td>
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			<td height="21" style="height:21px;width:216px;">Small tooth comb</td>
			<td style="width:232px;">Goody</td>
			<td colspan="2">No catalog number_ Goody Start.Style.Finish. Link to company website http://www.goody.com/Products/Styling_Tools/Goody_StartStyleFinish_Combs?colId=d11c04a1-1dd3-4848-a313-046fc6c109ec&#38;c_id=35a36c88-d359-4104-a53c-dfbfd8452780</td>
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			<td height="42" style="height:42px;width:216px;">Alcohol preps</td>
			<td style="width:232px;">Kendall Webcol Alcohol Preps</td>
			<td style="width:312px;">No catalog number. Kendall Webcol Alcohol Preps</td>
			<td>https://www.buyemp.com/product/kendall-webcol-alcohol-preps</td>
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			<td height="21" style="height:21px;width:216px;">Painter&#39;s tape</td>
			<td style="width:232px;">3M Scotch</td>
			<td>ScotchBlue ORIGINAL Painter&#8217;s Tape</td>
			<td>https://www.scotchblue.com/3M/en_US/scotchblue/products/painters-tape/</td>
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			<td height="21" style="height:21px;width:216px;">Permanent marker</td>
			<td style="width:232px;">Sharpie</td>
			<td style="width:312px;">Black Sharpie Fine Point</td>
			<td>http://www.sharpie.com/sharpie/fine/SHFineBlackFine.html</td>
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			<td height="42" style="height:42px;width:216px;">Non-latex gloves</td>
			<td style="width:232px;">Prot&#233;g&#233;</td>
			<td style="width:312px;">93997</td>
			<td style="width:479px;">Protege&#174;Nitrile Powder-Free Examination Gloves http://aureliagloves.com/product/protege/</td>
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			<td height="21" style="height:21px;width:216px;">Hair clips&#160;</td>
			<td style="width:232px;">Goody</td>
			<td>Goody Start.Style.Finish. Curl Clips</td>
			<td>http://www.goody.com/Products/Styling_Tools/Goody_StartStyleFinish_Curl_Clips</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Envelope</td>
			<td style="width:232px;">Office Depo</td>
			<td>330808</td>
			<td>Office Depot&#174; Brand Clasp Envelopes, 9&#34; x 12&#34;, Brown, Box Of 100</td>
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			<td height="21" style="height:21px;width:216px;">Foil sheets&#160;</td>
			<td style="width:232px;">Choice&#160;</td>
			<td>1229105</td>
			<td>Box of 500 Sheets Food Service Interfolded Pop-up foil sheets&#160;</td>
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		<tr height="21">
			<td height="21" style="height:21px;">Pre-weighed 2 ml cryovial</td>
			<td>Wheaton, Millville, NJ, USA</td>
			<td>W985902</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">100% isopropanol</td>
			<td style="width:232px;">Spectrum</td>
			<td style="width:312px;">I1275 | 67-63-0</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Liquid nitrogen&#160;</td>
			<td style="width:232px;">Ohio State Unversity Stores</td>
			<td>Gas Cylinder Warehouse</td>
			<td>http://osustores.osu.edu/prodservices/liquidnitrogen.aspx</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Stainless steel ball bearing&#160;</td>
			<td>McMasterCarr</td>
			<td>9291K42</td>
			<td>https://www.mcmaster.com/#standard-stainless-steel-balls/=1acqby0</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Specially milled aluminum cassettes</td>
			<td style="width:232px;">FisherScientific&#160;</td>
			<td style="width:312px;">Retsch 20.745.0001</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">HPLC grade methanol</td>
			<td style="width:232px;">FisherScientific&#160;</td>
			<td style="width:312px;">A452N1-19</td>
			<td>https://www.fishersci.com/shop/products/methanol-hplc-fisher-chemical-6/p-216129</td>
		</tr>
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			<td height="21" style="height:21px;">Microcentrifuge tube</td>
			<td style="width:232px;">Eppendorf North America</td>
			<td style="width:312px;">22363352</td>
			<td>https://online-shop.eppendorf.us/US-en/Laboratory-Consumables-44512/Tubes-44515/Eppendorf-Safe-Lock-Tubes-PF-8863.html?_ga=2.36544352.1776616592.1511283212-1254533852.1511283212</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Cortisol ELISA Kit</td>
			<td>Salimetrics LLC, State College, PA, USA</td>
			<td>1-3002 (5PK 1-3002-5)</td>
			<td>https://www.salimetrics.com/assay-kit/cortisol-salivary-elisa-eia-kit</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Centtrifuge for the micorcentrifuge tube</td>
			<td style="width:232px;">Beckman Coulter</td>
			<td style="width:312px;">Microfuge 20 series</td>
			<td>http://centrifugebybeckman.com/?page_id=727/?pi_ad_id=creative?utm_term=microfuge</td>
		</tr>
	</tbody>
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collecting hair samples for hair cortisol analysis in african americans

The hormone cortisol is typically assessed in saliva, serum, or urine samples. More recently, cortisol has been successfully extracted from hair, including humans. The advantage of hair cortisol concentration is that it reflects a retrospective representation of hypothalamic-pituitary-adrenal (HPA) axis function over time, much like hemoglobin A1C represents glycemic control. However, obtaining hair samples can be challenging, due to the cultural beliefs and hair care practices of minority participants. For example, African Americans may be reluctant to provide samples. Additionally, few researchers are trained to collect hair samples from African Americans. The purpose of this paper is to present a culturally informed protocol to help researchers obtain hair samples from African Americans. To illustrate the representative results of this protocol implementation, de-identified data from African Americans that participated in a community-based study on chronic stress are provided. Hair practice preferences are assessed. The participants are made comfortable by showing pictures of hair samples prior to cutting their hair. The single strain twist and gently pull method is used to collect approximately 30 - 50 strands of hair from the posterior vertex region of the scalp. This protocol will significantly improve collection of hair samples from African Americans.

In our study, twenty-one African Americans (mean age = 75; SD = 6.9; 85% female) consented to participate. Four participants (19%) refused to provide a hair sample and two (9.5%) could not provide a sample because their hair was too short. A total of 15 hair samples were obtained and processed (Table 2). The length of the hair samples that were processed ranged from 1 cm to 4.8 cm. Technical problems included samples were poorly aligned at the scalp end, tangled, or curly...

Watch this Scientific Journal Video about Collecting Hair Samples for Hair Cortisol Analysis in African Americans at JoVE.com

Collecting Hair Samples for Hair Cortisol Analysis in African Americans

Hair cortisol concentration analysis provides an alternative to traditional measures of cortisol; however, to collect hair samples from African Americans, scientists need to be culturally informed and competent. The purpose of this protocol is to demonstrate a culturally informed technique to collect hair samples for cortisol analysis from African Americans.

The hormone cortisol is typically assessed in saliva, serum, or urine samples. More recently, cortisol has been successfully extracted from hair, ...

This method can help researchers acquire adequate hair samples in a culturally sensitive manner. The main advantages of this technique are its culturally informed approach and the twist and roll maneuver that is used to secure the samples. Implementation of this technique will help increase the participation of African American subjects in studies about the influence of longterm cortisol exposure and health outcomes.Our protocol takes into account a careful consideration of physical, psychosocial, and cultural characteristics relevant to the collection of hair samples from the African American adults. Visual demonstration of this method is critical for appreciation of the importance of hair in the African American community and to provide the necessary steps for improving the participants experience. Before collecting the sample, explain to the participant that the sample collected is less hair than is lost in normal everyday brushing from the back of the head.Inform the participant how the site for the sampling is hidden by the surrounding hair and therefore not visible after collection, and that the sample will be used to measure a hormone called cortisol that is present in the hair. Show the participant a sample photo to illustrate the amount of hair that will be collected. Then, ask the participant the hair care practice questions listed in the table.Next, escort the participant to a private area. Don a pair of non-latex gloves, and clean a pair of salon grade scissors and a large and small tooth parting comb with an alcohol prep pad. Gently pull the hair in the sample taut to allow measurement from the scalp to the ends of the strands.Use the comb to part the hair horizontally along the posterior vertex of the scalp between the tips of the ears, and use a clip to secure the hair from the top of the part. Then, place another clip at the bottom of the part to expose a five by 10 centimeter rectangle between the two clips of loose hair, and ask the participant if they prefer a wide tooth or a narrow tooth comb. After combing the hair rectangle, ask the participant how they would like to have the remaining hair in the comb discarded.And, grasp approximately 30 to 50 strands of hair to the right of the five by 10 centimeter rectangle. Gently pull and twist the hair away from the scalp in a rolling motion between the fingers, and use the scissors to make a clean, straight cut with the root ends aligned carefully along the cut. Use painter's tape to secure the hair sample in the center of an eight by 10 inch piece of aluminum foil, and use a marker to label the scalp end of the sample.Fold the foil into a square so the hair cannot fall out of the foil, but without bending the hair. Place the foil into a business envelope. After sealing, use a premade barcode label to label the envelope with the participant's unique study identification number, and store the sample in a dry area at room temperature.If you do not have a premade barcode label, you can write the participant's unique id number, date, and hair sample on an envelope. The hair cortisol can then be analyzed according to standard protocols. 21 African Americans consented to participate in this representative study.Four participants ultimately refused to provide a hair sample, and two could not provide a sample because their hair was too short. A total of 15 usable hair samples with lengths ranging from one to 4.8 centimeters were obtained and processed from the other study participants. Technical problems included samples that were poorly aligned at the scalp end, too small of a sample, curly, or with painter's tape being over the scalp end.Once mastered, this technique can be completed in 15 minutes if performed properly. While attempting this procedure, it's important to remember to thank the participants and offer to give the participants any unused hair. Although thinning shears have been used in previous studies, shears are not the preferred tool for cutting hair because their use results in uneven scalp end alignment and an increased time for preparing the hair samples for analysis.This technique paved the way for researchers in the hair cortisol analysis field in obtaining culturally informed hair samples from African American study participants and in quantifying the potential influences of longterm cortisol exposure on health in African Americans. After watching this video, you should have a good understanding of how to approach and to collect adequate hair samples for cortisol concentration analysis from African American adults. Don't forget that working with salon grade scissors can be extremely sharp, so take care not to handle the blade ends of the scissors and not to press the scissors directly against the participant's scalp.

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