这出版物由克利夫兰的临床和翻译科学合作使成为可能;KL2TR000440 国家卫生研究院 (NCATS) 国家促进转化科学中心;和 NIH 的医学研究路线图。朱迪. 福特博士在这一领域的工作由国立国家药物滥用研究所 (1R21DA034960) 和俄亥俄州立大学人口研究研究所资助, 由国立儿童健康研究所和发展 (2P2CHD058484)。内容完全是作者的责任, 不一定代表国家卫生研究院的官方意见。Laudenslager 博士在这一领域的工作是由以病人为中心的结果研究所 (CE-1304-6208) 的一份合同提供资金的, 这是由儿童和家庭当局 (90YR0058) 提供的补助金, 以及发展 Psychobiology 的补助金。科罗拉多大学丹佛安舒茨医学院精神病学系研究组捐赠基金。

这项研究获得了克利夫兰医疗中心机构审查委员会的批准, 该议定书遵循了审查委员会所指出的参加者保护和数据安全准则。
1. 为头发样本和发样收集问题准备参与者
<ol><li>向参加者解释, 收集的毛发样本比从头部后部 (后顶点) 每天刷牙的头发少。告诉参与者, 头发样本的网站是由周围的头发隐藏, 因此, 收集后不可见。通知参与者, 样品将被用来测量一种叫做皮质醇的荷尔蒙, 它存在于头发10,11。</li>
	<li>取出样本照片, 并向参与者显示将收集的头发量 (30-50 股)。 注: 见图 1为头发样本的照片。</li>
	<li>询问参加者的头发护理实践12问题表 1中列出。</li>
</ol>2. 收集头发样本
<ol><li>告诉参与者, 样品是收集在一个私人区域在一个便携的分区, 窗帘, 或关闭门。向参与者保证, 如果他们戴上假发或假发, 他们的头发不会在别人看到的地方被剪掉。通过轻轻拉紧头发, 从头皮到末端, 测量头发长度。</li>
	<li>如果头发短 (小于3厘米), 按照步骤3中的短头发协议 (虽然头发不到1厘米长, 样品不能收集, 理想情况下, 头发应该至少有3厘米长)。对于长度超过6厘米的头发, 请按照步骤4中的长头发协议进行。否则, 请继续执行下一步。</li>
	<li>确保材料通过放置理发用品 (一张铝箔, 商业信封, 沙龙级剪刀, 大和小齿分梳, 酒精眼影, 画家胶带, 永久性标记, 非乳胶手套 [可选], 和头发) 做好准备。剪辑)10在参与者旁边的曲面上。在收集样品之前, 用酒精准备垫清洁剪刀和梳子。切出一个 8 x 10 "片铝箔用于样品。</li>
	<li>戴上非乳胶手套。把梳子和部分头发沿头皮的后顶点水平沿着耳朵的尖端。</li>
	<li>拿一个夹子, 把头发从零件的顶端移动, 然后在底部放置另一个剪辑, 在两个松散的头发 (大约是纸牌的大小) 之间暴露一个 5 x 10 厘米的矩形。</li>
	<li>询问参加者, 如果他们喜欢使用宽齿或窄齿梳通过 2-3 "矩形的头发。询问参与者, 她或他希望在梳子上留下的毛发被丢弃。把梳子上的头发给参与者, 或者, 如果 (而且只有当) 参与者说, 它可以扔掉松散的头发, 把剩下的头发从梳子扔进垃圾桶。</li>
	<li>抓住大约 30-50 股头发的权利, 2-3 "矩形, 并轻轻地拉和扭曲头发远离头皮在滚动运动之间的手指。</li>
	<li>确保样品收集尽可能接近头皮, 但不要削减头皮。使用沙龙级剪刀, 做一个干净的直切。</li>
	<li>抓住头发样本, 确保根端沿切口小心对齐。</li>
	<li>用画家的胶带把头发贴在铝箔纸上。不要覆盖头皮末端, 也不要在短于或等于3厘米的头发上使用画家的胶带. 将松散的剪发放到箔的中心, 折叠成一个正方形, 确保头发不脱落的箔。</li>
	<li>把标记和标签的头皮端在铝箔上。使用预制条码标签附加到箔和信封。</li>
	<li>折叠铝箔不折叠头皮端的头发, 以防止床头 (皱和弯曲的头发), 把它放在商业大小信封, 和印章。</li>
	<li>使用参与者唯一的研究标识号、日期和主要调查员的姓名, 将信封外部标上标签, 或者如果有的话, 可以用条形码标签。</li>
	<li>在室温下将样品贮存在干燥区域。</li>
</ol>3. 短头发标本收集 (1-3 厘米)
<ol><li>执行步骤2.3 和2.4。</li>
	<li>离别后, 请参加者把分开的头发紧贴头皮。剪头发沿部分, 同时用食指和拇指10紧紧地拿着头发。</li>
	<li>将松散的毛发放在箔上, 然后安全折叠。不要把头发贴在箔上。</li>
	<li>执行步骤2.11 至2.14。</li>
</ol>4. 长头发样本收集 (长于6厘米)
<ol><li>部分头发左到右在后顶点。</li>
	<li>使用发夹来保留额外的头发。</li>
	<li>创建一个扭曲的头发, 紧紧抓住食指和拇指, 以免下降样本10。</li>
	<li>使干净的直切尽可能靠近头皮。收集 30-50 股头发。</li>
	<li>如果参与者的头发很薄, 则在后顶点处切开 2-3 小面积 (1 厘米), 以隐藏切口的部位。</li>
	<li>执行步骤2.10 至2.14。</li>
</ol>5. 样品加工
<ol><li>将含有头发样品的箔包无限期存放在室温下直到分析。 注意: 头发皮质醇是非常稳定的头发, 可以储存多年。事实上, 从木乃伊标本中测定的皮质醇是11。</li>
	<li>分析以前发表的12的头发皮质醇。样品可以在常温下隔夜装运。</li>
</ol>

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行为免疫学和内分泌实验室被指定为 Salimetrics, 有限责任公司的卓越实验室。

头发收集协议的基本步骤包括提供一个例子, 说明将采取多少头发, 显示样本站点 (后涡), 使用单一的应变扭和轻轻拉法, 并确保样品与画家的磁带远离头皮末端。通过轻轻地扭和拉样品, 卷曲的头发被拉长, 头皮端对齐保持。这样可以减少对齐头皮末端、测量和切割样品所需的实验室时间。尽管在先前的研究中使用了变薄剪, 但由于头皮末端排列不均, 而且在准备头发样本进行分析时, 它们并不是切割毛发的首选工具。
收集来自非洲裔美国人的毛发样本对皮质醇的测量是可行的。在非洲裔美国人的文化中, 欣赏头发的意义是必要的, 以获得支持收集头发样本。调查人员不应该传达关于头发卫生的判断, 比如洗发的频率, 因为一些非洲裔美国人的头发比非西班牙裔白人的13还要干。非拉美裔白人 (M = 396 µm/天-1, SD = 55 µm/天-1)14, 非洲裔美国人的头发增长慢, 以平均率256µm/天-1 。参与者也可以佩戴发型, 如编织/缝在头发, 盒辫子, 和古怪的曲折或钩针辫子。这些发型需要数小时才能完成, 费用从50元到500美元不等。如果可能, 询问有关发型和协调头发样本收集与参与者。在这些情况下, 自然头发可能需要收集的美容师任命或沙龙。
研究者对头发、风格偏好和成本的重要性缺乏欣赏, 这可能导致参与者不愿提供头发样本。正如目前的研究所反映的那样, 非洲裔美国人不情愿或无力提供毛发样本的这种现象在多种族青少年的研究中被记录在案, 并且在皮质醇测量中收集毛发的可行性和可接受性8. 福特等。(2016) 抽取516名年龄在11-17 岁的混合种族青少年。非洲裔美国人青少年不太可能提供头发样品由于短发和拒绝 (或 = 0.26; 95% CI [0.13, 0.51])8。
本研究的参与者为65岁及以上。头发皮质醇分析的最大限制是剃光头, 这在男性或头发稀疏的人中很常见。收集足够的头发为样本在年长的成年人可能是挑战, 因为脱发。头发的重量更重要, 因为如果没有足够的质量, 那么微板块阅读器就不能准确地检测皮质醇水平。标准化的头发重量作为一个规范化的因素之前, 处理样品的皮质醇分析可能是一个未来的考虑, 以解决这一挑战。
在以前的头发皮质醇研究中已经概述了一些局限性, 包括头发护理实践的变化和皮质醇的标准值缺乏结果8,12,15。头发直、松弛剂、操控、调理剂和染料的使用改变了头发皮质醇的水平。暴露于漂白和操控减少皮质醇的数量, 而过氧化物增加测量皮质醇15。此外, 缺乏规范的价值提出了一个挑战, 比较头发皮质醇的结果跨研究, 以建立正常的价值观。我们研究的头发皮质醇的结果是有限的, 只能推广到其他老年人和慢性病患者。一般情况下, 慢性疾病 (糖尿病、心血管) 和情绪紊乱患者的头发皮质醇水平高于没有慢性病9、16的人。另一项对社区居住的非洲裔美国人老年人的研究报告说, 皮质醇的含量从0.93 到 6.36, 低于我们研究16的范围。另一项关于多民族老年心血管疾病样本的研究报告说, 皮质醇水平为 22.1 pg/毫克, 类似于本研究的结果。
本文所概述的技术的未来应用应包括教学参与者的捻度和轻轻拉程序, 使他们能够收集自己的头发样本。在非洲裔美国人的乳腺癌患者中进行了自我样本收集, 并被认为是更可接受的16。然而, 在乳腺癌研究中, 提供头发样本的非洲裔美国妇女的反应率很低。为了提高非洲裔美国人参与收集头发 biospecimens 的研究, 需要研究更好地了解拒绝的原因以及任何潜在的年龄或队列人口对拒绝率的影响。正如本研究中发现的那样, 以一种非批判性的方式接近潜在参与者, 承认在非洲裔美国人社区中毛发和毛发的重要性, 分享头发样本的照片, 提供隐私, 并通过扭曲和轻轻拉法是所有的因素, 归因于成功的参与者参加了研究。
目前, 没有发布的协议考虑文化偏好收集头发样本的皮质醇。同样, 很少有调查人员被训练来收集来自非洲裔美国人的头发。单应变扭和轻轻拉剪头发样本协议可以用于其他研究, 需要来自非洲裔美国人的头发样本。该议定书有可能增加来自非洲裔美国人的毛发样本数量, 以及有文化背景的调查人员和数据收集者的数量。

收集唾液, 血液, 或尿液的皮质醇分析有独特的程序性挑战和更大的参与者负担, 因为程序需要多个唾液样本, 静脉穿刺样本, 或24小时尿收集。从头发样本获得皮质醇是一种简单而可接受的替代方法, 使用唾液, 血清, 或尿样1,2。头发皮质醇提供了一个回顾性表示下丘脑-垂体-肾上腺 (HPA) 轴功能随着时间的推移。从头皮上收集的大约3厘米的毛发相当于过去3月1的平均皮质醇释放率。然而, 研究人员在尝试收集不同人群的毛发样本时遇到了独特的挑战。这些挑战包括宗教或精神信仰, 包括揭露头发 (如穆斯林妇女), 头发作为神圣的 (美洲印第安人团体), 或文化规范, 连接头发的标准的美丽和自我形象。
在非洲裔美国人的文化中, 尤其是在女性中, 头皮毛发是他们身体形象3,4的一个非常个人和重要的方面。此外, 历史经验 (例如, 塔斯基吉实验) 在研究过程5中继续助长不信任。一些非洲裔美国人的民俗和文化信仰也表明, 毛发样本可以用于可能造成伤害6,7的仪式。研究人员应该意识到, 当非洲裔美国人的参与者被要求提供毛发样本时, 他们可能面临的潜在挑战。
除了围绕身体图像的问题外, 收集和处理来自非洲裔美国人的毛发样本的具体技术可能与欧洲美国人6,7不同。例如, 非洲裔美国人可能有直, 薄, 卷曲, 古怪, 粗糙, 或这些毛发类型的组合, 每个需要不同的技术来收集和确保足够的头发样本处理。最近对非洲裔美国人青少年样本的研究8以及多种族样本9表明, 在非洲裔美国人的参与下, 为皮质醇水平测量收集毛发是可以接受的。目前, 尚无视觉训练程序, 旨在为研究人员提供在非洲裔美国人收集毛发样本的技能和文化知识。本文的目的是提供一个文化上知情的协议, 以帮助研究人员获得足够的头发样本来自非洲裔美国人的参与者。这一方法是在仔细考虑了与收集来自非洲裔美国人的毛发样本有关的生理、心理和文化特征时制定的。到目前为止, 还没有收集这些样品的视觉方法, 这项议定书将是第一个指南, 详细介绍了从非洲裔美国人成人正确的头发样本收集。为了说明《议定书》执行工作的代表性结果, 从非洲裔美国人那里得到了不确定的数据, 这些人参加了一项以社区为基础的关于65岁以上的慢性压力的研究。来自不同种族背景的男性和女性研究助理都与临床上产生的名单和传单的潜在参与者联系。主要调查员, 谁是非洲裔美国妇女, 在社区中心和高级补贴住房公寓向年长的成年非洲裔美国人介绍了这项研究, 包括对毛发收集方法的描述。

<table><tbody><tr><td>&amp;nbsp;Salon grade scissors</td><td>Cricket</td><td>S-3 550</td><td>Carded Professional Hair Shear, 5.5 inch</td></tr><tr><td>Large tooth comb</td><td>Goody</td><td>No catalog number__Goody Stylista Rake Comb&amp;nbsp;&amp;nbsp;&amp;nbsp;</td><td>Link to Company website&amp;nbsp; http://www.goody.com/Products/Styling_Tools/Goody_Stylista_Rake_Comb?colId=d11c04a1-1dd3-4848-a313-046fc6c109ec&amp;amp;c_id=35a36c88-d359-4104-a53c-dfbfd8452780</td></tr><tr><td>Small tooth comb</td><td>Goody</td><td>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&amp;amp;c_id=35a36c88-d359-4104-a53c-dfbfd8452780</td><td></td></tr><tr><td>Alcohol preps</td><td>Kendall Webcol Alcohol Preps</td><td>No catalog number. Kendall Webcol Alcohol Preps</td><td>https://www.buyemp.com/product/kendall-webcol-alcohol-preps</td></tr><tr><td>Painter&#039;s tape</td><td>3M Scotch</td><td>ScotchBlue ORIGINAL Painter&amp;rsquo;s Tape</td><td>https://www.scotchblue.com/3M/en_US/scotchblue/products/painters-tape/</td></tr><tr><td>Permanent marker</td><td>Sharpie</td><td>Black Sharpie Fine Point</td><td>http://www.sharpie.com/sharpie/fine/SHFineBlackFine.html</td></tr><tr><td>Non-latex gloves</td><td>Prot&amp;eacute;g&amp;eacute;</td><td>93997</td><td>Protege&amp;reg;Nitrile Powder-Free Examination Gloves http://aureliagloves.com/product/protege/</td></tr><tr><td>Hair clips&amp;nbsp;</td><td>Goody</td><td>Goody Start.Style.Finish. Curl Clips</td><td>http://www.goody.com/Products/Styling_Tools/Goody_StartStyleFinish_Curl_Clips</td></tr><tr><td>Envelope</td><td>Office Depo</td><td>330808</td><td>Office Depot&amp;reg; Brand Clasp Envelopes, 9&quot; x 12&quot;, Brown, Box Of 100</td></tr><tr><td>Foil sheets&amp;nbsp;</td><td>Choice&amp;nbsp;</td><td>1229105</td><td>Box of 500 Sheets Food Service Interfolded Pop-up foil sheets&amp;nbsp;</td></tr><tr><td>Pre-weighed 2 ml cryovial</td><td>Wheaton, Millville, NJ, USA</td><td>W985902</td><td></td></tr><tr><td>100% isopropanol</td><td>Spectrum</td><td>I1275 | 67-63-0</td><td></td></tr><tr><td>Liquid nitrogen&amp;nbsp;</td><td>Ohio State Unversity Stores</td><td>Gas Cylinder Warehouse</td><td>http://osustores.osu.edu/prodservices/liquidnitrogen.aspx</td></tr><tr><td>Stainless steel ball bearing&amp;nbsp;</td><td>McMasterCarr</td><td>9291K42</td><td>https://www.mcmaster.com/#standard-stainless-steel-balls/=1acqby0</td></tr><tr><td>Specially milled aluminum cassettes</td><td>FisherScientific&amp;nbsp;</td><td>Retsch 20.745.0001</td><td></td></tr><tr><td>HPLC grade methanol</td><td>FisherScientific&amp;nbsp;</td><td>A452N1-19</td><td>https://www.fishersci.com/shop/products/methanol-hplc-fisher-chemical-6/p-216129</td></tr><tr><td>Microcentrifuge tube</td><td>Eppendorf North America</td><td>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><td>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><td>Centtrifuge for the micorcentrifuge tube</td><td>Beckman Coulter</td><td>Microfuge 20 series</td><td>http://centrifugebybeckman.com/?page_id=727/?pi_ad_id=creative?utm_term=microfuge</td></tr></tbody></table>

collecting hair samples for hair cortisol analysis in african americans

荷尔蒙皮质醇通常在唾液、血清或尿样中进行评估。最近, 皮质醇已经成功地从毛发中提取, 包括人类。头发皮质醇浓度的优势在于它反映了随着时间的推移, 下丘脑-垂体-肾上腺 (HPA) 轴功能的回顾性表现, 就像血红蛋白 A1C 代表血糖控制。然而, 由于少数参与者的文化信仰和护发行为, 获得毛发样本可能具有挑战性。例如, 非洲裔美国人可能不愿意提供样品。此外, 很少有研究人员被训练来收集来自非洲裔美国人的毛发样本。本文的目的是提供一个文化上知情的协议, 以帮助研究人员获得来自非洲裔美国人的头发样本。为了说明本议定书执行工作的代表性结果, 提供了参加基于社区的关于慢性压力的研究的非裔美国人的数据。评估头发练习偏好。参加者通过在理发前展示头发样本的图片使他们感到舒适。单应变扭和轻轻拉法是用来收集大约 30-50 股头发从后顶点区域的头皮。这项议定书将大大改善来自非洲裔美国人的毛发样本的收集。

在我们的研究中, 二十一非洲裔美国人 (平均年龄 = 75;SD = 6.9;85% 女性) 同意参加。四名参加者 (19%) 拒绝提供头发样本, 两人 (9.5%) 不能提供样本, 因为他们的头发太短。共获得和处理15发样品 (表 2)。处理的毛发样本的长度从1厘米到4.8 厘米不等. 技术问题包括样品在头皮末端、纠结或卷曲, 并且画家的磁带被放置在头皮末端。
在实施单应变扭和轻轻拉的协议, 头发样本被排列和皮质醇含量确定。头发样品的头皮末端的近3厘米被切开, 洗涤和地面, 然后皮质醇提取与乙醇。根据制造商的指示, 使用商业高灵敏度 EIA 试剂盒确定头发皮质醇浓度 (HCC)。对以前的地面毛发进行了联合控制, 并在每一个 EIA 板上分别提取并包括在一起, 用于测定变异间的检测系数。所有示例都以重复的运行。对照毛池的变异系数 (CV) 小于 10%, 重复检测的内 cv 值小于4%。头发皮质醇浓度报告为 pg/毫克。平均皮质醇含量为 20.4 pg/毫克, 标准差为 22.7 pg/毫克, 结果从 4.3 pg/毫克到非规模, 甚至在额外稀释后 (图 2)。此外, 在实验室里处理样品的时间也少了。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57288/57288fig1.jpg"> 图 1。照片的头发样本, 表明放置画家的磁带上箔, 以确保样品.<a href="/files/ftp_upload/57288/57288fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57288/57288fig2.jpg"> 图 2。头发皮质醇浓度的直方图.结果分布不均匀。头发皮质醇的结果往往是扭曲的, 需要统计转换 (e., 日志转换), 以纠正不均匀分布的数据。<a href="/files/ftp_upload/57288/57288fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table fo:keep-together.within-page="1"><tbody>
<tr>
<td>告诉我你一周洗头发的次数？</td>
		</tr>
<tr>
<td>告诉我你一个月洗头发的次数？</td>
		</tr>
<tr>
<td>告诉我你一年洗头发的次数？</td>
		</tr>
<tr>
<td>你的头发上使用调理剂吗？ 是或否</td>
		</tr>
<tr>
<td>你的头发是颜色还是漂白？</td>
		</tr>
<tr>
<td>在过去的3月里, 你烫头发还是烫发？</td>
		</tr>
<tr>
<td>在过去的3月里, 你的头发上有化学矫直剂吗？</td>
		</tr>
<tr>
<td>你是否接管了任何头皮条件的柜台或处方药？</td>
		</tr>
<tr>
<td>你在过去的3月里服用过类固醇吗？例如, 吸入药物, 在关节内注射类固醇, 或其他口服类固醇, 如泼尼松, 松, 地塞米松, 或醛固酮。</td>
		</tr>
</tbody></table>表1。可能会影响头发中皮质醇浓度的毛发护理的清单.例如, 巴西头发矫直的做法减少了头发中皮质醇的含量。
<table fo:keep-together.within-page="1"><tbody>
<tr>
<td>头发重量 (毫克)</td>
			<td>皮质醇 (pg/毫克)</td>
			<td>头发长度</td>
			<td>颜色</td>
			<td>剪发技术</td>
			<td>评论</td>
		</tr>
<tr>
<td>9.15</td>
			<td>10。3</td>
			<td>3厘米</td>
			<td>黑白相间</td>
			<td>细化剪</td>
			<td>头发没有排列。</td>
		</tr>
<tr>
<td>5.91</td>
			<td>46。0</td>
			<td>3厘米</td>
			<td>金发和黑色的。切两个。</td>
			<td>细化剪</td>
			<td>头发没有排列。头发纠结, 所以只采取了头发, 可以确定为头皮末端。</td>
		</tr>
<tr>
<td>5.02</td>
			<td>13。7</td>
			<td>3厘米</td>
			<td>黑白相间</td>
			<td>细化剪</td>
			<td>头发没有排列。头发纠结, 所以只采取了头发, 可以确定为头皮末端。</td>
		</tr>
<tr>
<td>30。9</td>
			<td>30。2</td>
			<td>1.4 厘米</td>
			<td>黑白相间</td>
			<td>细化剪</td>
			<td>头发放在信封里没有锡纸。很难收集头发。</td>
		</tr>
<tr>
<td>31.07</td>
			<td>6。7</td>
			<td>1厘米</td>
			<td>黑白相间</td>
			<td>细化剪</td>
			<td>头发放在信封里没有锡纸。很难收集头发。</td>
		</tr>
<tr>
<td>6.45</td>
			<td>12。7</td>
			<td>3.3 厘米</td>
			<td>黑白相间</td>
			<td>沙龙级剪刀</td>
			<td>头发非常卷曲。把头发剪得尽可能接近3厘米。</td>
		</tr>
<tr>
<td>6.13</td>
			<td>105。2</td>
			<td>3厘米</td>
			<td>黑白相间</td>
			<td>细化剪</td>
			<td>头发没有排列。头发纠结, 所以只采取了头发, 可以确定为头皮末端。</td>
		</tr>
<tr>
<td>6.81</td>
			<td>23。5</td>
			<td>3.2 厘米</td>
			<td>黑白相间</td>
			<td>沙龙级剪刀和扭拉</td>
			<td>头发非常卷曲。把头发剪得尽可能接近3厘米。</td>
		</tr>
<tr>
<td>4.94</td>
			<td>6。2</td>
			<td>3厘米</td>
			<td>黑</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>11。7</td>
			<td>10。5</td>
			<td>2.8 厘米</td>
			<td>黑白相间</td>
			<td>沙龙级剪刀和扭拉</td>
			<td>头发非常卷曲。把头发剪得尽可能接近3厘米。</td>
		</tr>
<tr>
<td>14</td>
			<td>4。3</td>
			<td>3.1 厘米</td>
			<td>黑色和白色的金发小贴士。切所有。</td>
			<td>沙龙级剪刀和扭拉</td>
			<td>头发非常卷曲。把头发剪得尽可能接近3厘米。</td>
		</tr>
<tr>
<td>29.08</td>
			<td>非规模</td>
			<td>4.8 厘米</td>
			<td>黑白相间</td>
			<td>细化剪</td>
			<td>头发楔在两片胶带之间。无法辨认头皮侧所以采取了整个长度。</td>
		</tr>
<tr>
<td>15.02</td>
			<td>8。4</td>
			<td>3厘米</td>
			<td>黑色带红色的紫色和白色的头皮。切所有。</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>18.25</td>
			<td>16。0</td>
			<td>3厘米</td>
			<td>黑</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>16.42</td>
			<td>29。5</td>
			<td>3厘米</td>
			<td>白色与一些淡褐色</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>32.93</td>
			<td>14。6</td>
			<td>3厘米</td>
			<td>黑色和灰色</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>29。9</td>
			<td>13。5</td>
			<td>3厘米</td>
			<td>灰色与一些淡褐色和黑色</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>28.89</td>
			<td>36。8</td>
			<td>3厘米</td>
			<td>灰色</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>22.42</td>
			<td>7。8</td>
			<td>3厘米</td>
			<td>黑</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>6.04</td>
			<td>23。6</td>
			<td>3厘米</td>
			<td>灰色</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
<tr>
<td>30.47</td>
			<td>5。0</td>
			<td>3厘米</td>
			<td>黑色与一些白色和棕色</td>
			<td>沙龙级剪刀和扭拉</td>
			<td></td>
		</tr>
</tbody></table>表2。头发皮质醇浓度的结果.表中的每条水平线代表一个参与者。第一列是在 mg 的头发重量, 其次是在样品中的 pg/毫克的皮质醇浓度。样品被测量, 并且长度在第三列从左边被表明了。一个参与者使用4.8 厘米长的头发, 因为样本量有限。与会者的头发颜色和对样品的评论也列出。

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

收集非洲裔美国人的头发样本进行头发皮质醇分析医学|视频

Collecting Hair Samples for Hair Cortisol Analysis in African Americans

头发皮质醇浓度分析提供了替代传统的皮质醇措施;然而, 要收集来自非洲裔美国人的毛发样本, 科学家需要有文化上的了解和能力。本议定书的目的是展示一种文化上消息灵通的技术来收集来自非洲裔美国人的皮质醇分析的毛发样本。

在非洲裔美国人中收集头发皮质醇分析样本

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

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17.9K 次观看.  The Ohio State University. 头发皮质醇浓度分析提供了替代传统的皮质醇措施;然而, 要收集来自非洲裔美国人的毛发样本, 科学家需要有文化上的了解和能力。本议定书的目的是展示一种文化上消息灵通的技术来收集来自非洲裔美国人的皮质醇分析的毛发样本。

视频: Collecting Hair Samples for Hair Cortisol Analysis in African Americans

A Modified Trier Social Stress Test for Vulnerable Mexican American Adolescents

The Trier Social Stress Test (TSST) is a well validated and widely used social stressor that has been shown to induce a 2-4 fold increase in cortisol, the biological output produced by the Hypothalamic-Pituitary-Adrenal (HPA) axis in humans. While studies have explored how modifications to the TSST influence stress responsivity, few studies have created a modified TSST appropriate for vulnerable youth that elicits a significant cortisol stress response. Thus, the current study sought to modify or adjust the TSST in a culturally sensitive way for a vulnerable sample of 14 year-old adolescents. The present study took place within the context of a longitudinal birth cohort study of Mexican American families in California called the Center for the Health Assessment of Mothers and Children of Salinas (CHAMACOS). The CHAMACOS sample was optimal to test the effectiveness of a modified culturally appropriate TSST, as it is comprised of Mexican American youth, who are often excluded from research. These youths also have experienced significant early life adversity. Example modifications included timed prompts, alternative math tasks, use of same-ethnicity peers as confederates, debriefing immediately after the conclusion of the TSST, and using an unknown youth examiner to administer the debrief. Saliva samples were collected at baseline (after a resting phase), and then again at 15, 30, and 45 min post-TSST onset to assess cortisol concentration. A pilot study of 50 participants (50% female) have been analyzed for cortisol reaction to the TSST. Results confirmed that this modified version of the TSST was successful at eliciting a significant cortisol reaction, with a wide range of variability likely due to individual differences. Goals for modifications and ethnicity considerations are discussed. This study provides the foundation for future research to utilize a modified TSST with vulnerable youth.

Here, we present a protocol that provoked cortisol reactivity in a vulnerable adolescent Mexican American sample utilizing a modified version of the Trier Social Stress Test (TSST). Saliva samples were collected at baseline, 15, 30, and 45 min post-TSST onset. Future research could utilize this modified TSST with vulnerable youth.

易受伤害的墨西哥美国青少年的改良特里尔社会压力测试

The Trier Social Stress Test (TSST) is a well validated and widely used social stressor that has been shown to induce a 2-4 fold increase in cortisol, the ...

科研

发育生物学

Collecting And Measuring Wound Exudate Biochemical Mediators In Surgical Wounds

We describe a methodology by which we are able to collect and measure biochemical inflammatory and nociceptive mediators at the surgical wound site. Collecting site-specific biochemical markers is important to understand the relationship between levels in serum and surgical wound, determine any associations between mediator release, pain, analgesic use and other outcomes of interest, and evaluate the effect of systemic and peripheral drug administration on surgical wound biochemistry. This methodology has been applied to healthy women undergoing elective cesarean delivery with spinal anesthesia. We have measured wound exudate and serum mediators at the same time intervals as patient's pain scores and analgesics consumption for up to 48 hours post-cesarean delivery. Using this methodology we have been able to detect various biochemical mediators including nerve growth factor (NGF), prostaglandin E2 (PG-E2) substance P, IL-1&#x03B2;, IL-2, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-17, TNF&alpha;, INF&#x03B3;, G-CSF, GM-CSF, MCP-1 and MIP-1&#x03B2;. Studies applying this human surgical wound bioassay have found no correlations between wound and serum cytokine concentrations or their time-release profile (J Pain. 2008; 9(7):650-7).1 We also documented the utility of the technique to identify drug-mediated changes in wound cytokine content (Anesth Analg 2010; 111:1452-9).2

This article provides a detailed and visual description of a methodology for collecting and measuring biochemical inflammatory and nociceptive mediators at the surgical wound site following cesarean delivery. This human bioassay has been used to determine correlations between wound and serum cytokine concentrations and drug-mediated changes in wound cytokines, chemokines and neuropetides.

手术伤口伤口渗出液生化调解员在收集和测量

We describe a methodology by which we are able to collect and measure biochemical inflammatory and nociceptive mediators at the surgical wound site. ...

医学

Cortisol Measurement in Koala (Phascolarctos cinereus) Fur

Optimal methods of hormone extraction used to measure stress in animals across sample types are not always the same. Australia&#39;s iconic marsupial species, the koala (Phascolarctos cinereus), faces prolonged exposure to anthropogenic-induced stressors and assessment of chronic stress in wild populations is urgently warranted. One of the most effective ways to measure chronic stress is through analyzing the glucocorticoid hormone cortisol in hair or fur, as it supports physiological and behavioral responses. This laboratory validation study aims to test current techniques to validate an optimal hormone extraction method to be used as a non-invasive measure of cortisol in koala fur. It is recognized that using non-invasive techniques to measure stress hormones is preferred over traditional, invasive techniques due to their ideal practical and ethical standpoints. Additionally, it is comparatively easier to acquire fur from koalas than it is to acquire samples of their blood. This study used samples of koala fur acquired from the Adelaide Koala and Wildlife Hospital to run a number of hormone extraction techniques in an attempt to validate an optimal cortisol extraction method. Results showed that 100% methanol provided the most optimal solvent extraction compared to 100% ethanol or 100% isopropanol based on parallelism results. In conclusion, this method of cortisol extraction from koala fur provided a reliable non-invasive assay that could be used to study chronic stress in koalas.

Cortisol Measurement in Koala Phascolarctos cinereus Fur

We present a protocol to determine the optimal extraction solvent to measure cortisol from koala fur. The solvents used in this protocol are methanol, ethanol and isopropanol. Determining an optimal extraction solvent will aid in reliably measuring fur to determine the impact of chronic stress on koalas.

科蒂醇测量在考拉 （菲斯科拉托斯西尼鲁斯） 毛皮

Optimal methods of hormone extraction used to measure stress in animals across sample types are not always the same. Australia&#39;s iconic marsupial ...

生物化学

Cortisol Extraction from Sturgeon Fin and Jawbone Matrices

The aims of this study were to develop a technique for the extraction of cortisol from sturgeon fins using two washing solvents (water and isopropanol) and quantify any differences in fin cortisol levels among three main sturgeon species. Fins were harvested from 19 sacrificed sturgeons including seven beluga (Huso huso), seven Siberian (Acipenser baerii), and five sevruga (A. stellatus). The sturgeons were raised in Iranian farms for 2 years (2017-2018), and cortisol extraction analysis was conducted in South Korea (January-February 2019). Jawbones from five H. huso were also used for cortisol extraction. Data were analyzed using the general linear model (GLM) procedure in the SAS environment. The intra- and inter-assay coefficients of variation were 14.15 and 7.70, respectively. Briefly, the cortisol extraction technique involved washing the samples (300 &#177; 10 mg) with 3 mL of solvent (ultrapure water and isopropanol) twice, rotation at 80&#160;rpm for 2.5 min, air-drying the washed samples at room temperature (22-28 &#176;C) for 7 days, further drying the samples using a bead beater at 50 Hz for 32 min and grinding them into powder, applying 1.5 mL methanol to the dried powder (75 &#177; 5 mg), and slow rotation (40 rpm) for 18 h at room temperature with continuous mixing. Following extraction, samples were centrifuged (9,500 x g for 10 min), and 1 mL supernatant was transferred into a new microcentrifuge tube (1.5 mL), incubated at 38 &#176;C to evaporate the methanol, and analyzed via enzyme-linked immunosorbent assay (ELISA). No differences were observed in fin cortisol levels among species or in fin and jawbone cortisol levels between washing solvents. The results of this study demonstrate that the sturgeon jawbone matrix is a promising alternative stress indicator to solid matrices.

In this study, we present a protocol for cortisol extraction from the fin and jawbone of sturgeon species. Fin and jawbone cortisol levels were further examined by comparing two washing solvents followed by ELISA assays. This study piloted the feasibility of jawbone cortisol as a novel stress indicator.

皮质醇从鱼翅和爪骨基质提取

The aims of this study were to develop a technique for the extraction of cortisol from sturgeon fins using two washing solvents (water and isopropanol) ...

环境科学

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations.

We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.

Salivary measures have emerged in bio-behavioral research  that are easy-to-collect, minimally invasive, and relatively inexpensive  biologic markers of ...

Providing Meaningful Environmental Enrichment and Measuring Saliva Cortisol in Pigs Housed on Slatted Flooring

As pork is the most consumed meat worldwide, the welfare of animals in the swine industry has increasingly become a major public concern, which imposes a substantial pressure coming from customers, legislators and other stakeholders, to make management changes to improve the well-being of these animals. Several studies have demonstrated that providing environmental enrichment to pigs allows them to express their natural behavior, such as rooting and exploring, as well as nesting prior to farrowing, and is associated with reduced stress and improved production and welfare. However, many considerations should be taken into account when providing environmental enrichment, such as the type of floor, drainage and sewage systems, the pigs&#39; stage in life, the material, as well as its hanging method, height and location within the pen. The objectives of this paper are (1) to give methodologic information on how to provide a relatively simple and practical meaningful environmental enrichment for pigs which are housed on slatted floors during the different stages of their life, and (2) to demonstrate how to collect saliva samples for the measurement of cortisol concentrations, as a biomarker for acute stress. Protocols include information regarding the use of jute, cotton ropes, straw in racks, as well as chewable silicone sticks devices as environmental enrichment in pens of farrowing and lactation, weaners and finishers. In addition, the use of cotton rope for a non-invasive saliva samples collection for cortisol concentrations analysis is detailed. The protocols provided are relevant for professionals aiming to improve and monitor animal welfare, in both research and industrial swine farming.

This protocol demonstrates how to provide practical meaningful environmental enrichment for pigs which are housed on slatted flooring during the different stages of their lives, and how to collect saliva samples in a non-invasive manner for the measurement of cortisol concentrations, as a biomarker for acute stress.

提供有意义的环境富集和测量唾液皮质醇在猪安置在板状地板

As pork is the most consumed meat worldwide, the welfare of animals in the swine industry has increasingly become a major public concern, which imposes a ...

行为学

Human Circadian Phenotyping and Diurnal Performance Testing in the Real World

In our continuously developing &#39;around the clock&#39; society, there is a need to increase our understanding of how changes in biology, physiology and psychology influence our health and performance. Embedded within this challenge, is the increasing need to account for individual differences in sleep and circadian rhythms, as well as to explore the impact of time of day on performance in the real world. There are a number of ways to measure sleep and circadian rhythms from subjective questionnaire-based methods to objective sleep/wake monitoring, actigraphy and analysis of biological samples. This paper proposes a protocol that combines multiple techniques to categorize individuals into Early, Intermediate or Late circadian phenotype groups (ECPs/ICPs/LCPs) and recommends how to conduct diurnal performance testing in the field. Representative results show large differences in rest-activity patterns derived from actigraphy, circadian phase (dim light melatonin onset and peak time of cortisol awakening response) between circadian phenotypes. In addition, significant differences in diurnal performance rhythms between ECPs and LCPs emphasizes the need to account for circadian phenotype. In summary, despite the difficulties in controlling influencing factors, this protocol allows a real-world assessment of the impact of circadian phenotype on performance. This paper presents a simple method to assess circadian phenotype in the field and supports the need to consider time of day when designing performance studies.

Here, we present a method to investigate diurnal rhythms in performance following accurate categorization of participants into circadian phenotype groups based on the Munich ChronoType Questionnaire, gold standard circadian phase biomarkers and actigraphic measures.

现实世界中的人类昼夜Phenotyping和二年性能测试

In our continuously developing &#39;around the clock&#39; society, there is a need to increase our understanding of how changes in biology, physiology and ...

A Noninvasive Hair Sampling Technique to Obtain High Quality DNA from Elusive Small Mammals

Noninvasive genetic sampling approaches are becoming increasingly important to study wildlife populations. A number of studies have reported using noninvasive sampling techniques to investigate population genetics and demography of wild populations1. This approach has proven to be especially useful when dealing with rare or elusive species2. While a number of these methods have been developed to sample hair, feces and other biological material from carnivores and medium-sized mammals, they have largely remained untested in elusive small mammals. In this video, we present a novel, inexpensive and noninvasive hair snare targeted at an elusive small mammal, the American pika (Ochotona princeps). We describe the general set-up of the hair snare, which consists of strips of packing tape arranged in a web-like fashion and placed along travelling routes in the pikas&#x2019; habitat. We illustrate the efficiency of the snare at collecting a large quantity of hair that can then be collected and brought back to the lab. We then demonstrate the use of the DNA IQ system (Promega) to isolate DNA and showcase the utility of this method to amplify commonly used molecular markers including nuclear microsatellites, amplified fragment length polymorphisms (AFLPs), mitochondrial sequences (800bp) as well as a molecular sexing marker. Overall, we demonstrate the utility of this novel noninvasive hair snare as a sampling technique for wildlife population biologists. We anticipate that this approach will be applicable to a variety of small mammals, opening up areas of investigation within natural populations, while minimizing impact to study organisms.

We present a noninvasive sampling approach to efficiently collect hair samples from elusive small mammals, as shown for the American pika. We demonstrate the utility of this method by extracting DNA from sampled hair and amplifying several types of molecular markers commonly used in studies of wildlife ecology and conservation.

一种无创性的发采样技术，以获得难以捉摸的小型兽类高质量的DNA

Noninvasive genetic sampling approaches are becoming increasingly important to study wildlife populations. A number of studies have reported using ...

生物学

Extraction and Analysis of Cortisol from Human and Monkey Hair

The stress hormone cortisol (CORT) is slowly incorporated into the growing hair shaft of humans, nonhuman primates, and other mammals. We developed and validated a method for CORT extraction and analysis from rhesus monkey hair and subsequently adapted this method for use with human scalp hair. In contrast to CORT &#34;point samples&#34; obtained from plasma or saliva, hair CORT provides an integrated measure of hypothalamic-pituitary-adrenocortical (HPA) system activity, and thus physiological stress, during the period of hormone incorporation. Because human scalp hair grows at an average rate of 1 cm/month, CORT levels obtained from hair segments several cm in length can potentially serve as a biomarker of stress experienced over a number of months.
In our method, each hair sample is first washed twice in isopropanol to remove any CORT from the outside of the hair shaft that has been deposited from sweat or sebum. After drying, the sample is ground to a fine powder to break up the hair&#39;s protein matrix and increase the surface area for extraction. CORT from the interior of the hair shaft is extracted into methanol, the methanol is evaporated, and the extract is reconstituted in assay buffer. Extracted CORT, along with standards and quality controls, is then analyzed by means of a sensitive and specific commercially available enzyme immunoassay (EIA) kit. Readout from the EIA is converted to pg CORT per mg powdered hair weight. This method has been used in our laboratory to analyze hair CORT in humans, several species of macaque monkeys, marmosets, dogs, and polar bears. Many studies both from our lab and from other research groups have demonstrated the broad applicability of hair CORT for assessing chronic stress exposure in natural as well as laboratory settings.

Cortisol (CORT) accumulates in the growing hair shaft of humans and nonhuman primates. We describe methods for extracting and analyzing hair CORT with high precision and sensitivity. Measurement of hair CORT is particularly well-suited for assessing chronic stress over periods of weeks to months.

提取与分析皮质醇从人与猴的毛

The stress hormone cortisol (CORT) is slowly incorporated into the growing hair shaft of humans, nonhuman primates, and other mammals. We developed and ...

Fecal Glucocorticoid Analysis: Non-invasive Adrenal Monitoring in Equids

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, corticotrophin releasing hormone (CRH) is released from the hypothalamus in the brain. This stimulates the release of adrenocorticotrophic hormone (ACTH) from the pituitary gland, which in turn stimulates release of glucocorticoids from the adrenal gland. In horses the glucocorticoid corticosterone is responsible for several adaptations needed to support equine flight behaviour and subsequent removal from the aversive situation. Corticosterone metabolites can be detected in the feces of horses and assessment offers a non-invasive option to evaluate long term patterns of adrenal activity. Fecal assessment offers advantages over other techniques that monitor adrenal activity including blood plasma and saliva analysis. The non-invasive nature of the method avoids sampling stress which can confound results. It also allows the opportunity for repeated sampling over time and is ideal for studies in free ranging horses. This protocol describes the enzyme linked immunoassay (EIA) used to assess feces for corticosterone, in addition to the associated biochemical validation.

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. The method offers a non-invasive option to assess long term patterns in both domestic and free ranging horses. This protocol describes the enzyme linked immunoassay involved and the associated biochemical validation.

糖皮质激素粪分析:在马属动物无创监测肾上腺

Adrenal activity can be assessed in the equine species by analysis of feces for corticosterone metabolites. During a potentially aversive situation, ...

在非洲裔美国人中收集头发皮质醇分析样本

摘要

探索更多视频

易受伤害的墨西哥美国青少年的改良特里尔社会压力测试

手术伤口伤口渗出液生化调解员在收集和测量

科蒂醇测量在考拉（菲斯科拉托斯西尼鲁斯）毛皮

皮质醇从鱼翅和爪骨基质提取

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

提供有意义的环境富集和测量唾液皮质醇在猪安置在板状地板

现实世界中的人类昼夜Phenotyping和二年性能测试

一种无创性的发采样技术，以获得难以捉摸的小型兽类高质量的DNA

提取与分析皮质醇从人与猴的毛

糖皮质激素粪分析:在马属动物无创监测肾上腺

在非洲裔美国人中收集头发皮质醇分析样本

摘要

探索更多视频

易受伤害的墨西哥美国青少年的改良特里尔社会压力测试

手术伤口伤口渗出液生化调解员在收集和测量

科蒂醇测量在考拉 （菲斯科拉托斯西尼鲁斯） 毛皮

皮质醇从鱼翅和爪骨基质提取

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

提供有意义的环境富集和测量唾液皮质醇在猪安置在板状地板

现实世界中的人类昼夜Phenotyping和二年性能测试

一种无创性的发采样技术，以获得难以捉摸的小型兽类高质量的DNA

提取与分析皮质醇从人与猴的毛

糖皮质激素粪分析:在马属动物无创监测肾上腺

科蒂醇测量在考拉（菲斯科拉托斯西尼鲁斯）毛皮