儿童健康的尤尼斯·肯尼迪·施莱佛国立:这项工作是由工作，家庭和健康网络（www.WorkFamilyHealthNetwork.org），它是通过美国国立卫生研究院和美国疾病控制和预防了合作协议，资助支持与人类发展（格兰特＃U01HD051217，U01HD051218，U01HD051256，U01HD051276），国家老龄问题研究所（格兰特＃U01AG027669），行为科学和科学研究，国家心脏，肺和血液研究所（R01HL107240）和国家职业办公室安全与健康（格兰特＃U01OH008788，U01HD059773）。从威廉T格兰特基金会，阿尔弗雷德·P斯隆基金会和政府儿童和家庭补助金提供了额外资金。本出版物的内容完全是作者的责任，并不一定代表这些机构和办事处的官方意见。

下面的协议已被批准适用的机构审查委员会，并根据赫尔辛基宣言进行。 1。开始之前:标准Precuations 确保所有的样品采集工作人员已通过美国NIOSH要求的血源性病原体（BBP）的培训。 <ol><li> 工程控制 <ol><li>使用食品和药物管理局批准处置锐器容器来处理使用过的无菌，一次性使用采血针的。使用正确标记生物废料桶进来有可能感染物质接触的任何其他材料。 </li><li>不要装得过满锐器容器。更换容器，以防止受伤之前只能填写一个锐器容器到3/4的容量。有用于对每一个尖锐物品容器隔室充满度的一个指标。 </li><li>只有在锐器容器处置锐器。请勿使用尖锐物品容器作OT她浪费。 </li><li>处置进来的适当标记生物废料桶与潜在的BBP接触任何非清晰的材料。 </li></ol></li><li> 个人防护装备（PPE） <ol><li>始终正确使用个人防护装备，以防止感染从BBP。收集星展期间使用个人防护装备，包括手套，实验室外套，保护眼睛，长裤和封闭趾鞋。 </li><li>戴上手套来处理血液或任何有接触血液或其他潜在感染性材料（OPIMs）。手套不宜太紧也不能太松，因为这可能会妨碍"挤奶"（后述）的过程。不戴戒指，同时收集血点，因为它们可能刺破手套。 </li><li>穿白大褂，以保护皮肤和衣服从血液和OPIMs。白大褂已经进来了BBP接触，应在清洗重新使用之前10％的漂白剂/水溶液。不管曝光后，白大褂应定期清洗，以保持它彗星瘦。 </li><li>穿封闭趾鞋来保护脚对血液和OPIMs可能的接触。 </li><li>如果需要，戴护目镜，以防止血液和因飞溅OPIMs可能的接触。 </li><li>为了保持工作区的洁净度，在离开工作区域，以避免污染实验室区域以外的项目中删除所有个人防护装备。 </li></ol></li><li> 工作实践控制 <ol><li>之前和之后与每个参与者的身体接触，用肥皂和温水，持续20秒洗手。用毛巾关闭自来水。如果一个接收器无法使用，至少使用60％酒精的洗手液。 </li><li>不管之前消毒剂应用和删除的手套后洗手，用酒精洗手的机会。使用足够的消毒剂，以弥补双手的所有表面，用手在一起，直到干燥。不要冲洗或擦拭手用毛巾擦干，因为这会降低消毒剂的功效。 </li><li>不要吃或饮用或使用化妆品或隐形眼镜在血液采集领域。个人接触到BBPS可能发生通过这些路径是直接路由到粘膜。 </li><li>不处理生物危险品不戴手套，不处理无害材料与先前触及生物危险品手套。 </li></ol></li><li> 适当的家政服务 良好的内务管理将继续帮助防止暴露于BBP和防止交叉污染环境。 <ol><li>使用杀病毒微生物的消毒湿巾正确洁净区星展收集过程中的污染。 </li><li>保持生物危害或污染的废物分开的普通垃圾，污染的材料是那些有可见的血液或OPIM他们。 </li></ol></li><li> 曝光响应 如果暴露于BBP，响应于以下方式。 <ol><li>用肥皂和水立即彻底冲洗接触面积。快速动作可减少感染的风险。 </li><li>识别风险的来源和途径，以便能够说明情况的现场主管和/或响应医师。 </li><li>暴露发生后立即通知待命的主管。它强调以获得支持，现场主管，了解风险水平进行感染，因此如果能及时的医疗处理是必要的评估。在特定的曝光控制计划（执委会）制定了指导方针，必须提供给所有那些在暴露于BBP危险。 </li></ol></li></ol> 2。收集室和安装材料<ol><li>选择一个样本采集位置是不适合吃，如茶水间，那就是自由的所有不相关的项目设备。一个不想要在一个领域是拥挤，有便于移动是很重要的。下面介绍如何设置干血点集合（LAB）的区域。 </li><li>建立生物危害的废物桶和一个生物危害垃圾袋行内部。废物桶应是可访问的，同时坐下。 </li><li>用消毒湿巾或类似的产品彻底清洁收集区。处理用过的清洁实验室区域中的生物危害废料桶的所有材料。在此步骤中，应戴上手套，以保护皮肤免受暴露于有害化学物质。 </li><li>更换手套和建立收集剩余物资。 <ol><li>地方手套和酒精洗手液的收集表。 </li><li>将锐器容器内收集的椅子的地方。该锐器容器应带手套处理在任何时候，然后妥善处置时充满。 </li><li>将所有在收集区保护焊要求星展收集剩余的材料。包括滤纸采集卡，几个采血针，酒精棉片，纱布垫，以及创可贴，但不能太多，因此该地区混乱。做没有设立超过了可能被用于每个参与者。 </li></ol></li></ol> 3。星展样品Cellection 始终佩戴个人防护装备（实验室外套，手套，长裤，封闭趾鞋，如果需要，安全眼镜）收集过程中，并把保护焊（CHUX）在膝部保护自己的街头服装。 <ol><li> 准备参加: <ol><li>获得同意后，要求参加者，如果他/她会喜欢使用左手或右手的收集和选择一个手指的集合。避免严重老茧或损坏（撕裂）手指和任何环形的手指。典型的小指和大拇指未被选择，以便分别减少参与者的疼痛和促进集合。的指尖的侧面被用作收集地点的一致性和便于收藏。 </li><li>有参与者用手揉搓在一起，温暖他们的手和刺激血液流动。一只手娃RMER可以使用，或者参与者可以运行他们的手温开水送下，如果接收器是可用的。另一种技术来刺激血液流动将有参与者交替握紧和放松的目标手，然后指尖前休息充分。 </li><li>清洁所选手指用酒精垫并使其充分干燥。与第二个酒精棉片重复完全去除任何污染物或洗手液。手指必须完全干燥在进行手指棒或酒精之前，将污染的血液样本。与所有用于收集材料，立即在生物危险废物容器处理的酒精擦拭。 </li></ol></li><li> 准备穿刺部位上的参与者的手指: <ol><li>确保参与者的手臂，以防止收集过程中的运动和放松手臂和手部肌肉。休息参与者的手肘在桌子的边缘，或在他们的膝盖上。保持参与者的手POSitioned舒适低于心脏水平和他们的肌肉放松，促进血液流动。 </li><li>抓住拇指和食指之间的选择的手指，慢慢牢牢挤压手指移动时从手掌的把握，以指尖血拉向指尖轻轻牛奶手指。 </li><li>按微柳叶刀坚决反对参与者的手指垂直于指纹放电柳叶刀。不刺破手指或接近甲床的中心，这两个点之间而不是目的，在手指的尺侧。在锐器容器立即丢弃用过的柳叶刀的。 </li></ol></li><li> 采血点: <ol><li>擦拭的第一滴血了，用无菌纱布垫除去样品中的组织液，并立即处置纱布进入生物危害的废物桶。 </li><li>同时保持参与者的手低于其心脏有向下的角度，根TLY奶手开始在手腕上下共同努力，手指，产生血液的基础。不要只挤在手指上，因为这将减少血流量。牛奶中的手指，直到出现大幅回落是挂从指尖。 </li><li>收集使用集合文件斑点。 <ol><li>小心地将手指下方的收集纸张并允许下降到属于它自己的重量。这使得收集的最大体积血点。如果血池在滤纸上， 即 "珠"，轻轻拍打卡，以鼓励血液浸透进纸。 </li><li>如果血液过于粘稠，或滴太小，属于自己的体重，缓慢将收集到的纸张与血滴接触通过毛细作用吸收血液从手指不接触手指。如果手指触摸收集纸，移动到该集合区被破坏的文件的另一个地方。参与者的手指笑ULD从来没有接触滤纸。 </li><li>不要在彼此顶部血滴，即使血迹未填充集合圈。 </li><li>不要让血跑下来的手指或到指甲，因为这使得集合的大斑点更加困难。 </li></ol></li><li>如果参与者的血液在收集过程中血块的任何时间，请求允许刺另一根手指，然后清洁和包扎初始穿刺部位，并重新开始，如果该请求被接受。 <ol><li>为了防止凝血，继续挤奶参与者的手指，以保持血液流动，或尝试牢牢擦拭穿刺用无菌纱布破坏血凝块形成。 注:后续手指棒可能需要几个原因:寒冷的房间可能会导致更快速凝固;还参加者的血液黏度可能不理想，或贫穷挤奶技术也会影响血流量。手暖，或温暖的自来水，用于PUNC前ture可反向寒冷的房间的效果。 </li></ol></li><li>一旦星展已收集，或者如果参与者不希望以后的手指棒，清理任何多余的血液穿刺部位。 </li><li>包扎穿刺部位，以防止潜在的污染，一旦参与者离开样品采集区。 </li></ol></li><li> FI质量控制（QC）的评估: 在完成样品的采集，评估的样品质量。确保从收集区的背面测量的斑点是1）至少0.375的直径; 2）通过收集纸充分浸湿，以便同样大小的滤纸上的两侧，斑点不符合该标准，表明通过手指到集合纸张接触并且容易涂抹集合; 3）不与彼此接触（重叠斑点的区域是不可测定的）。 </li><li> 完成样品的收集 <ol><li>以下GUI在步骤1.1中所述delines，处置任何剩余的污染收集的材料，包括可能一直在与潜在传染性物质接触任何物品。 </li><li>站上的保护焊的收集纸张干燥。空间收集卡片，让他们之间有良好的空气流通。的时间进行空气干燥的最小量为15分钟，优选至少30分钟。允许最大空气流量，折叠收藏卡的背面在一个方式，它可以自由地站立和提供有关收集纸样品背后气流是很重要的。 </li><li>所有步骤完成后，取出个人防护装备。 </li></ol></li><li> 响应对象的感觉淡淡的，或不能很好地 尽管可能性不大，参与者可能期间或之后的指尖采血变得头晕，眼花，或模糊。要警惕的迹象，如面色苍白，出汗的脸和额头，视力模糊，下垂或眼睑飘飘的投诉，或恶心的投诉。 T的发病灰褐色的症状往往是突然的，样品收集器必须准备迅速作出反应，如果任何的症状是可见的。如果任何这些症状都存在在任何点的过程中，根据下面的程序进行反应。 <ol><li>如果参与者是感觉淡淡的，眼花，头晕，或出现昏厥的迹象，采取以下措施: <ol><li>停止该过程。 </li><li>要小心，参与者不属于或成为受伤。 </li><li>平静地安抚参与者，必要时，要求参加者以弯曲的腰，把他/她的头他/她的两腿之间。如果流动性是有限的，参与者可以移动，相邻的缓冲位置可以用来休息。 </li><li>有参加者休息10分钟。 </li><li>恢复过程，如果参与者同意继续。 </li></ol></li><li>如果参与者昏倒，采取以下措施: <ol><li>停止该过程。 </li><li>要小心，参与者不属于或成为受伤。 </li><li>有与会者趴在他/她的背部尽可能快地用双脚抬高。参与者将被指示直接从坐姿躺下没有站起来。 </li><li>请参与者松开任何过紧的衣服。 </li><li>有参加者休息10分钟。 </li><li>血片采集不应该继续。 </li><li>如参加者没有1分钟后回应，拨打911。 </li></ol></li></ol></li><li> 打破实验室面积: 一旦血点的收集工作已经完成，收集区域必须被分解和净化清楚地让过路人的区域不会被感染BBP。采血区，设备容器，收集材料和废弃物的容器必须干净BBP的。与往常一样，处理潜在传染性物质时戴上个人防护装备。 <ol><li>从收集区取出并重新包装所有未受污染的材料。 </li><li>处理任何剩余的污染废物材料进入生物废料桶。 </li><li>关闭该尖锐物品容器，并将其存储在提供保护免受污染的区域。如果锐器容器是3/4满，适当地处理它，并更换新的锐器容器。 </li><li>请用杀病毒微生物的消毒湿巾整个样本收集区。 </li><li>一旦所有的生物危险品已被出售，生物危害袋可高压灭菌。当从垃圾筒取出袋子，谨慎，只摸袋的外面，并封住包装。这也是很好的做法，使用杀病毒微生物湿巾清洁所有设备的外部，以防止交叉污染。 </li></ol></li></ol> 4。培训，收集星展银行验证任何干血片采集器是由合格的个人进行多项式前已杜热的病人或研究对象。确保收藏者展示尊重参与者的信心和熟练使用采血用品和个人防护装备的，有足够的容量点的集合中，并在完成过程中的及时和有效的方式，最好在10分钟。 5。样品包装与贮存 <ol><li>确保DBS样品已经干燥至少15分钟，它们被装入袋中存放前，饱和卡可能需要30分钟或更长的时间来充分地干燥。关闭采集卡盖，确保样品完全干燥，防止拖尾和污染样品。 </li><li>采取生物危害标本袋中，然后放入干燥剂小袋放入标本袋。将星展样品进入生物危害标本袋。总是处理星展样品时戴上手套。不要让血液样本接触到样品袋的外侧。 </li><li>将每个样品在它自己的生物危害样品袋机智哈哈干燥剂包。压平包以除去多余的空气，然后将袋子密封严密。将袋装样品在储存冷却器用于运输或临时存储。避免高温。 </li><li> 2周集合内冻结或测定DBS样品，如DBS的样本通常可以在室温下储存两周无显著降解取决于分析物7,2。 </li></ol> 6。样品运输和货运 <ol><li>除非已知含有传染性病原体，船舶干血斑样本作为下运输法规美国能源部，并按照第2，第6章豁免人类标本，§的国际民用航空组织（ICAO）的技术指南8 6.3.2.3.6 。 </li><li>如果冻，船样过夜干冰与有害物质所需的额外的预防措施，或gelpaks冷冻到-80°C。船舶上周一至周三为了避免解冻在运输过程中因延误或周末/节假日，与确认接收站点，他们期待并准备好接收和冷冻样品。 </li></ol> 7。质控考核<ol><li>以下收集和之前测定（以最小化冻融循环的次数优选冻结前）评估样品的质量。 </li><li>包括样品质量的评估下列条件，适当的包装，颜色编码的干燥剂保持完整（干），斑点的饱和度，斑点的大小，适当的收集方法包括点不重叠或涂在卡指示与主体手指接触。 </li></ol> 8。冷冻贮藏虽然降解时间不同的分析物通过，冻结DBS样品2周集合内以获得最佳效果。冷冻样品与干燥剂在实验室级的冷冻机中。如同任何生物试样，最小化冻融Çycles，尽管DBS的样本中的一些试验已经表明没有证据表明退化的高达6次冻融循环9-11。

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Chem. 55, 1034-1036 (2009).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=International+Civil+Aviation+Organization..">International Civil Aviation Organization..</a> Technical Instructions for the Safe Transport of Dangerous Goods by Air. , (2010).</li><li>McDade, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epstein-Barr+virus+antibodies+in+whole+blood+spots:+a+minimally+invasive+method+for+assessing+an+aspect+of+cell-mediated+immunity.">Epstein-Barr virus antibodies in whole blood spots: a minimally invasive method for assessing an aspect of cell-mediated immunity.</a> Psychosom. Med. 62, 560-567 (2000).</li><li>McDade, T. W., Burhop, J., Dohnal, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-sensitivity+enzyme+immunoassay+for+C-reactive+protein+in+dried+blood+spots.">High-sensitivity enzyme immunoassay for C-reactive protein in dried blood spots.</a> Clin. Chem. 50, 652-654 (2004).</li><li>McDade, T. W., Shell-Duncan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole+blood+collected+on+filter+paper+provides+a+minimally+invasive+method+for+assessing+human+transferrin+receptor+level.">Whole blood collected on filter paper provides a minimally invasive method for assessing human transferrin receptor level.</a> J. Nutr. 132, 3760-3763 (2002).</li><li>Gerszten, R. E., Wang, T. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+search+for+new+cardiovascular+biomarkers.">The search for new cardiovascular biomarkers.</a> Nature. 451, 949-952 (2008).</li><li>D'Agostino, R. B., Grundy, S., Sullivan, L. M., Wilson, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Validation+of+the+Framingham+coronary+heart+disease+prediction+scores:+results+of+a+multiple+ethnic+groups+investigation.">Validation of the Framingham coronary heart disease prediction scores: results of a multiple ethnic groups investigation.</a> J. Am. Med. Assoc. 286, 180-187 (2001).</li><li>McDade, T. W., Rutehrford, J., Adair, L., Kuzawa, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+origins+of+inflammation:+microbial+exposures+in+infancy+predict+lower+levels+of+C-reactive+protein+in+adulthood.">Early origins of inflammation: microbial exposures in infancy predict lower levels of C-reactive protein in adulthood.</a> Proc. R Soc. B.. , (2009).</li><li>McDade, T. W., Ice, G. H., James, G. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Measuring Stress in Humans. A Practical Guide for the Field. , 181-207 (2007).</li><li>Tanner, S., McDade, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Enzyme+immunoassay+for+total+immunoglobulin+E+in+dried+blood+spots.">Enzyme immunoassay for total immunoglobulin E in dried blood spots.</a> Am. J. Hum. Biol. 19, 440-442 (2007).</li><li>Miller, A. A., Sharrock, K. C., McDade, T. W. <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+leptin+in+dried+blood+spot+samples.">Measurement of leptin in dried blood spot samples.</a> Am. J. Hum. Biol. 18, 857-860 (2006).</li><li>Miller, E. M., McDade, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+highly+sensitive+immunoassay+for+interleukin-6+in+dried+blood+spots.">A highly sensitive immunoassay for interleukin-6 in dried blood spots.</a> Am. J. Hum. Biol. , (2012).</li></ol>

奥尔菲巴克斯顿已经收到了研究者发起的补助金从Sepracor公司公司（现Sunovion; ESRC-0977，ClinicalTrials.gov标识符NCT00900159），作为一个顾问和专家证人斯莫尔有限责任公司，于松谷美的科学顾问委员会，并获得咨询费用从维克森林大学医学中心。

生物标志物是跨多个学科的非常有用的工具，包括医学，细胞生物学，遗传学，心理学，社会学，人口学，社会流行病学。在每一个学科，早期阶段的有效性和可靠性的生物标志物的研究被用来揭示两个隐藏的疾病的风险，并提供健康结果的纵向预测。在这些情况下，生物标志物，是准确的，耐用的（ 即它们可以存活样本收集和实施实验室测定之间的滞后，提取该生物标记物），和通用（ 即有用的各种型号）是必需的。 而星展银行的集合是一个相对简单的过程，有几个技巧可以帮助最大限度地采集点数量。之前收集开始，确保参与者的手是温暖的，因此该血管扩张，有利于血液流动，一旦手指被刺破。用暖手器，或具有参与者执行他们的手温开水送下可以帮助这一进程。挤奶手指集合之前会拉血的指尖和温暖的手，帮助血液流动。挤奶是一个过程，实践与提高。收藏家应该使用什么技术，他们发现最舒适，最有效的，但最常见的方法包括挤奶参与者的手与收藏家的另一只手。这种方法将允许收藏家的拇指工作深入到参与者的手掌，而另一只手可以稳定参与者的手腕上。作为一个例子，如果一个人从手指上的参与者的左手采血，收集器将稳定在参与者的手腕的人用左手和乳用右手。而挤奶包括不使用严格控制，上面拉的手，而工作到肌肉拉血往指尖的皮制成常见的错误。同样重要的是避免销清手指与挤奶手或紧紧地攥着参与者的手腕与nonmilking手，这两个阻碍血液流动。 当穿刺手指，使穿刺尽可能深地牢牢地压入柳叶刀放电前的手指。浅穿刺导致更少的血流量和更快凝固。刺破手指后，很快工作是关键。薄血导致更容易收集和较大的斑点。穿刺后，使血液变得越来越粘稠，直到凝固。收集血点的卡就越大其体积所需的时间就越少。 理想情况下，参与者的血液会瘦，以至于将下降属于自己的体重，但这不会是对所有参与者的情况下，有些人的血更厚或比别人快凝块。如果血太厚从手指回落，过滤卡必须被带到血液采集。瓦EN使卡到其血液中的下降联系人持有该卡，而它被挂在手指，只要有可能，而不是只涂抹血液到卡上是很重要的。这将有助于拉血液从手指以最大化光点的大小。这种技术需要稳定的双手，以维持血液和卡之间的接触。用双手，一是稳住参与者的手和一个持有该卡，并保持双手互相接触，有助于卡和稳定的血液​​一起。 先进的研究领域，如心脏代谢风险评估，长期依赖生物标志物如C-反应蛋白（CRP），胰岛素，糖化血红蛋白（HbA1c）的早期风险检测和预防工作，以及"硬终点"的生物标志物对于心血管疾病（CVD）的风险和心血管疾病相关的死亡12。临床流行病学经常评估生物标志物来prospectivelŸ预测心脏代谢风险的结果，如心脏疾病和II型糖尿病。也许是最广泛使用的所有生物标记物是Framingham风险分数，复合，包括高血压，高密度脂蛋白胆固醇水平和总胆固醇的水平，已证明在预测CVD 13效用。然而，Framingham评分经历了一些技术性的争议，并有一定的局限性的研究主要集中在社会和环境因素链接到健康，提示需要涉及到心血管研究更准确，更具体的生物标志物。 有许多使用ELISA或可​​供DBS样品的测定法类似的技术可商购的试剂盒。必要DBS样品的测定是使用特定的和优化的洗脱步骤，使干燥的样品在溶液中2。这是研究的一个扩展区域为目前正在开发用于检测多种被分析物1。中日已经验证，被广泛用的电子检测是胆固醇2，CRP 10,14,15，IgE16，EB病毒9,15，17瘦素和cyotkines 18。有许多专门的实验室在星展银行的分析，其中包括华盛顿大学实验室医学系（UW医学实验室）生物标志物实验室（导演:马克雯儿）的。 铭记生物标志物组成的小组，而不是一个孤独的生物标志物，是最有用的心血管疾病和糖尿病研究领域，生物标志物需要具有各种背景因素，以最大限度地发挥其效用要考虑沿。生物标志物的研究可能很昂贵但是，包括财政和数据收集时间。因此，尽管它是非常重要的选择生物标志物，是准确，耐用，用途广泛，工作和家庭的生物标志物的研究必须植根于实用性通过选择生物标志物，价格低廉，适合，并能迅速收集的Wi个放心。此外，实际的考虑往往导致无法在一个程序，同时最大限度地提高价格，质量和速度。在这里躺下场和人口研究中的医疗生物标志物研究的下一个步骤。生物标志物研究领域目前正处于飞速发展阶段，但是，大部分的工作还在后头。

生物标志物可以提供关于生物过程，从正常到致病性可能有助于临床上可识别的疾病1有价值的信息。在研究环境中，干燥血点（DBS）正变得越来越优选以静脉穿刺来采集生物标志物在全血中的方法。 DBS集合是简单的，相对无痛，微创然后静脉穿刺，并且需要最小的存储要求（ 即样品不需要被立即冻结，可存放相当长的时间周期在测定之前稳定冷冻环境）。 DBS样品可用于各种不同的分析物，包括胆固醇，C-反应蛋白，糖化血红蛋白，Epstein-Barr病毒的抗体，和一些细胞因子的检测。 静脉穿刺长期以来一直是黄金标准的血液生物标志物的集合。但是，获取，存储和装运血样需要adherenCE到多种协议和健康的代码，这可能无法在某些研究协议。抽血，必须静脉穿刺正式认证，而星展银行的抽样程序更简单，不需要官方认证。静脉采血还需要更多的设备比不收集星展银行（ 即针，管，止血带），多收集后的努力。静脉穿刺样品必须进行处理（ 即血清或血浆通过离心或在室温下静置对给定的时间期间内从血液样品中提取的）以及在-20℃和-70℃冷冻Ç立即以防止分析物的降解而DBS样品必须被简单地在空气中干燥，然后可在室温下保存一周或更依赖于分析物，转移至冷冻室2之前。星展样品收集更便宜比静脉穿刺样本进行上述原因。星展样品也占用空间少，降低了运输和存储成本之前检测。 利用唾液作为生物标志物已成为关键的生物行为研究。唾液提供了一个简单，非侵入性的方法来收集生物标记物，已成为现场工作，旨在测量皮质醇或丁宁3-5的标准。尿液是另一个试样，可以无创地进行收集，并且可以用在群体水平上研究褪黑激素的测定或用于毒理学。虽然这两种尿和唾液提供了简单的是能够通过参与者自身完成采集程序，不是所有的分析物可以在这些介质进行测量。 星展提供了一个简单，具成本效益，替代静脉穿刺，并在该领域日益成为实施的研究工具和人口研究6。

<table border="0" cellpadding="0" cellspacing="0" width="573">
	<colgroup>
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	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Asepti II Microbial Wipes</td>
			<td style="width:71px;">EcoLab</td>
			<td style="width:119px;">D12330345</td>
			<td style="width:167px;">3rd party shipping</td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Bio-hazard waste bags</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">14220-092</td>
			<td style="width:167px;">High Strength Autoclavable Polypropylene Biohazard Bags, 2 mil</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;"></td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Bio-hazard waste barrel</td>
			<td style="width:71px;">Tyco</td>
			<td style="width:119px;">58578-092</td>
			<td style="width:167px;">8g. Slide lid</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Sharps Container</td>
			<td style="width:71px;">BD Medical</td>
			<td style="width:119px;">BD305487</td>
			<td style="width:167px;">1.4 qt.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Chux</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">82020-845</td>
			<td style="width:167px;">Absorbent bench underpads&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Gloves</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">XSmall: 89038-267</td>
			<td style="width:167px;">Powder-free, latex-free nitrile gloves</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;"></td>
			<td style="width:71px;"></td>
			<td style="width:119px;">Small: 89038-268</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;"></td>
			<td style="width:71px;"></td>
			<td style="width:119px;">Medium: 89038-270</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;"></td>
			<td style="width:71px;"></td>
			<td style="width:119px;">Large: 89038-272</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;"></td>
			<td style="width:71px;"></td>
			<td style="width:119px;">XLarge: 89038-274</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Hand sanitizer (e.g., Purell)</td>
			<td style="width:71px;">Johnson &#38; Johnson</td>
			<td style="width:119px;">89040-210</td>
			<td style="width:167px;">3rd party shipping</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Alcohol wipes</td>
			<td style="width:71px;">WEBCOL and CURITY</td>
			<td style="width:119px;">82003-856</td>
			<td style="width:167px;">Prep pads</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;"></td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Gauze</td>
			<td style="width:71px;">VERSALON</td>
			<td style="width:119px;">82004-826</td>
			<td style="width:167px;">Non-woven sponges</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;"></td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Band-Aids</td>
			<td style="width:71px;">Nutramax Stat Strips</td>
			<td style="width:119px;">15431-475</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Lancets</td>
			<td style="width:71px;">Owen Mumford</td>
			<td style="width:119px;">82028-060</td>
			<td style="width:167px;">3.0 mm, 21 ga.&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Protein paper collection cards</td>
			<td style="width:71px;">Whatman</td>
			<td style="width:119px;">Custom-build</td>
			<td style="width:167px;">903 Protein Saver Collection paper</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Desiccant</td>
			<td style="width:71px;">VWR</td>
			<td style="width:119px;">61161-319</td>
			<td style="width:167px;">Humidity sponges</td>
		</tr>
		<tr height="168">
			<td height="168" style="height:168px;width:216px;">Specimen bags</td>
			<td style="width:71px;">Lab Guard Reclosable Biohazard Bags, ITW Minigrip</td>
			<td style="width:119px;">56766-370</td>
			<td style="width:167px;">6&#8221; x 9&#8221;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;"></td>
			<td style="width:71px;"></td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Specimen storage cooler</td>
			<td style="width:71px;">Igloo</td>
			<td style="width:119px;">Playmate Mini</td>
			<td style="width:167px;">4 qt.; direct from manufacturer</td>
		</tr>
	</tbody>
</table>

dried blood spot collection of health biomarkers to maximize participation in population studies

生物标记是疾病，健康，曝光，或其他生物体信息的直接测量的生物学指标。在人口和社会科学，生物标志物必须是容易获得，运输，并分析。干血斑满足这种需要，并且可以被收集在该领域具有高的响应率。这些元素是特别重要的在纵向研究设计，包括干预措施，其中流失是至关重要的，以避免和高反应率提高结果的解释。干血斑样本采集简便，快捷，相对无痛，创伤小，然后静脉穿刺，并且仅需最少的领域存储需求（ 即样品不需要立即冷冻，可以保存的很长一段时间之前，在一个稳定的环境下冷冻测定法）。该样品可用于各种不同的分析物，包括胆固醇，C-反应蛋白，糖化血红蛋白，众多的细胞因子和o的被分析疗法的分析物，以及提供遗传物质。星展集合被描绘成用在最近的几项研究。

<img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/50973/50973fig1highres.jpg" src="/files/ftp_upload/50973/50973fig1.jpg" /> 图1。干血斑样本，一个采集卡显示，在干燥过程中的四个，有效的收集点。 <a href="https://www.jove.com/files/ftp_upload/50973/50973fig1highres.jpg" target="_blank">点击这里查看大图。</a> <img alt="图2" fo:content-width="5in" fo:src="/files/ftp_upload/50973/50973fig2highres.jpg" src="/files/ftp_upload/50973/50973fig2.jpg" /> 图2:好的和坏的干血片样本。成功（右）和不成功（左）星展集合结果显示， <a href="https://www.jove.com/files/ftp_upload/50973/50973fig2highres.jpg" target="_blank">点击此处查看大IM</a>年龄。 <img alt="图3" fo:content-width="5in" fo:src="/files/ftp_upload/50973/50973fig3highres.jpg" src="/files/ftp_upload/50973/50973fig3.jpg" /> 图3。经理的态度和与员工心血管代谢疾病的风险6相关的做法，这个数字已经被修改伯克曼，LF，巴克斯顿，OM，厄特尔，K.＆奥科丘克沃，工作-家庭平衡有关C.经理的行为预测员工的心血管风险和睡眠持续时间延长的医疗环境。 中国职业健康心理学 115，316-329（2010年）。暴露在管理者与开放性和创造力，帮助其员工的工作和家庭需要的中等或低工作与家庭的成绩的员工，有具有两个或多个心血管疾病的风险（目前吸烟者，肥胖，高血压或炒作的统计学显著增加的可能性rtension，高胆固醇和高糖化血色素（HbA1c）或糖尿病的诊断）。顶部和中间板描绘出平均值±标准偏差为总胆固醇由干血片和血红蛋白A1C由医疗点设备（也可以通过干血斑可靠地计量）的评估综合评估。底部面板由经理得分显示心血管疾病的风险。分析仅限于护士和其他员工提供直接护理表明，暴露于低创造性和开放性管理有关的工作，家庭问题可能是特别重要的，这些人（或低WF得分= 6.4，P = 0.01）。所有型号控制了年龄，性别，工资，教育，种族/民族，和工地。 <a href="https://www.jove.com/files/ftp_upload/50973/50973fig3highres.jpg" target="_blank">点击这里查看大图。</a>

Watch this Scientific Journal Video about Dried Blood Spot Collection of Health Biomarkers to Maximize Participation in Population Studies at JoVE.com

Dried Blood Spot Collection of Health Biomarkers to Maximize Participation in Population Studies

生物标志物是疾病或健康的直接测量的生物指标。在人口和社会科学，生物标志物必须是容易获得，运输，并分析。干血斑（DBS）的集合满足这方面的需求，可以在现场收集与高响应速率和多种生物标志物分析。

干生生物标志物的血斑收藏最大限度地参与人口研究

Biomarkers are directly-measured biological indicators of disease, health, exposures, or other biological information. In population and social sciences, ...

dried-blood-spot-collection-health-biomarkers-to-maximize

Research

JoVE Journal

Medicine

36.2K 次观看.  Harvard School of Public Health. 生物标志物是疾病或健康的直接测量的生物指标。在人口和社会科学，生物标志物必须是容易获得，运输，并分析。干血斑（DBS）的集合满足这方面的需求，可以在现场收集与高响应速率和多种生物标志物分析。 

视频: Dried Blood Spot Collection of Health Biomarkers to Maximize Participation in Population Studies

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

在动物模型揭示神经，血液，创伤后应激障碍和行为相关因素的生物标志物

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

科研

医学

High Throughput Sequential ELISA for Validation of Biomarkers of Acute Graft-Versus-Host Disease

Unbiased discovery proteomics strategies have the potential to identify large numbers of novel biomarkers that can improve diagnostic and prognostic testing in a clinical setting and may help guide therapeutic interventions. When large numbers of candidate proteins are identified, it may be difficult to validate candidate biomarkers in a timely and efficient fashion from patient plasma samples that are event-driven, of finite volume and irreplaceable, such as at the onset of acute graft-versus-host disease (GVHD), a potentially life-threatening complication of allogeneic hematopoietic stem cell transplantation (HSCT).
Here we describe the process of performing commercially available ELISAs for six validated GVHD proteins: IL-2R&alpha;5, TNFR16, HGF7, IL-88, elafin2, and REG3&alpha;3 (also known as PAP1) in a sequential fashion to minimize freeze-thaw cycles, thawed plasma time and plasma usage. For this procedure we perform the ELISAs in sequential order as determined by sample dilution factor as established in our laboratory using manufacturer ELISA kits and protocols with minor adjustments to facilitate optimal sequential ELISA performance. The resulting plasma biomarker concentrations can then be compiled and analyzed for significant findings within a patient cohort. While these biomarkers are currently for research purposes only, their incorporation into clinical care is currently being investigated in clinical trials.
This technique can be applied to perform ELISAs for multiple proteins/cytokines of interest on the same sample(s) provided the samples do not need to be mixed with other reagents. If ELISA kits do not come with pre-coated plates, 96-well half-well plates or 384-well plates can be used to further minimize use of samples/reagents.

High throughput validation of multiple candidate biomarkers can be performed by sequential ELISA in order to minimize freeze/thaw cycles and use of precious plasma samples. Here, we demonstrate how to sequentially perform ELISAs for six different validated plasma biomarkers1-3 of graft-versus-host disease (GVHD)4 on the same plasma sample.

高通量顺序酶联免疫吸附试验的急性移植物抗宿主病的生物标志物的验证

Unbiased discovery proteomics strategies have  the potential to identify large numbers of novel biomarkers that can improve  diagnostic and prognostic ...

An Affordable HIV-1 Drug Resistance Monitoring Method for Resource Limited Settings

HIV-1 drug resistance has the potential to seriously compromise the effectiveness and impact of antiretroviral therapy (ART). As ART programs in sub-Saharan Africa continue to expand, individuals on ART should be closely monitored for the emergence of drug resistance. Surveillance of transmitted drug resistance to track transmission of viral strains already resistant to ART is also critical. Unfortunately, drug resistance testing is still not readily accessible in resource limited settings, because genotyping is expensive and requires sophisticated laboratory and data management infrastructure. An open access genotypic drug resistance monitoring method to manage individuals and assess transmitted drug resistance is described. The method uses free open source software for the interpretation of drug resistance patterns and the generation of individual patient reports. The genotyping protocol has an amplification rate of greater than 95% for plasma samples with a viral load &#62;1,000 HIV-1 RNA copies/ml. The sensitivity decreases significantly for viral loads &#60;1,000 HIV-1 RNA copies/ml. The method described here was validated against a method of HIV-1 drug resistance testing approved by the United States Food and Drug Administration (FDA), the Viroseq genotyping method. Limitations of the method described here include the fact that it is not automated and that it also failed to amplify the circulating recombinant form CRF02_AG from a validation panel of samples, although it amplified subtypes A and B from the same panel.

Drug resistance testing for HIV-1 infected individuals failing antiretroviral therapy (ART) can guide future therapies and improve treatment outcomes. Optimizing individual and population health outcomes in high HIV prevalence but resource-limited settings will ultimately require affordable and accessible drug resistance genotyping and interpretation methods.

对于资源有限设置一个负担得起的HIV-1耐药性监测方法

HIV-1 drug resistance has the potential to seriously compromise the effectiveness and impact of antiretroviral therapy (ART). As ART programs in ...

Fundus Photography as a Convenient Tool to Study Microvascular Responses to Cardiovascular Disease Risk Factors in Epidemiological Studies

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an important role in maintaining cardiovascular health. The retina is a tissue that lines the interior of the eye and it is the only tissue that allows for a non-invasive analysis of the microvasculature. Nowadays, high-quality fundus images can be acquired using digital cameras. Retinal images can be collected in 5 min or less, even without dilatation of the pupils. This unobtrusive and fast procedure for visualizing the microcirculation is attractive to apply in epidemiological studies and to monitor cardiovascular health from early age up to old age.
Systemic diseases that affect the circulation can result in progressive morphological changes in the retinal vasculature. For example, changes in the vessel calibers of retinal arteries and veins have been associated with hypertension, atherosclerosis, and increased risk of stroke and myocardial infarction. The vessel widths are derived using image analysis software and the width of the six largest arteries and veins are summarized in the Central Retinal Arteriolar Equivalent (CRAE) and the Central Retinal Venular Equivalent (CRVE). The latter features have been shown useful to study the impact of modifiable lifestyle and environmental cardiovascular disease risk factors.
The procedures to acquire fundus images and the analysis steps to obtain CRAE and CRVE are described. Coefficients of variation of repeated measures of CRAE and CRVE are less than 2% and within-rater reliability is very high. Using a panel study, the rapid response of the retinal vessel calibers to short-term changes in particulate air pollution, a known risk factor for cardiovascular mortality and morbidity, is reported. In conclusion, retinal imaging is proposed as a convenient and instrumental tool for epidemiological studies to study microvascular responses to cardiovascular disease risk factors.

Retinal image analysis is an unobtrusive procedure for visualizing the microcirculation. The impact of cardiovascular disease risk factors can result in changes of retinal vessel calibers. The procedures to acquire fundus images and the steps for calculating the vessel calibers are described.

眼底摄影作为一种方便的工具，研究微血管回应流行病学研究心血管疾病的风险因素

The microcirculation consists of blood vessels with diameters less than 150 &#181;m. It makes up a large part of the circulatory system and plays an ...

Quantification of the Immunosuppressant Tacrolimus on Dried Blood Spots Using LC-MS/MS

The calcineurin inhibitor tacrolimus is the cornerstone of most immunosuppressive treatment protocols after solid organ transplantation in the United States. Tacrolimus is a narrow therapeutic index drug and as such requires therapeutic drug monitoring and dose adjustment based on its whole blood trough concentrations. To facilitate home therapeutic drug and adherence monitoring, the collection of dried blood spots is an attractive concept. After a finger stick, the patient collects a blood drop on filter paper at home. After the blood is dried, it is mailed to the analytical laboratory where tacrolimus is quantified using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) in combination with a simple manual protein precipitation step and online column extraction.
For tacrolimus analysis, a 6-mm disc is punched from the saturated center of the blood spot. The blood spot is homogenized using a bullet blender and then proteins are precipitated with methanol/0.2 M ZnSO4 containing the internal standard D2,13C-tacrolimus. After vortexing and centrifugation, 100 &#181;l of supernatant is injected into an online extraction column and washed with 5 ml/min of 0.1 formic acid/acetonitrile (7:3, v:v) for 1 min. Hereafter, the switching valve is activated and the analytes are back-flushed onto the analytical column (and separated using a 0.1% formic acid/acetonitrile gradient). Tacrolimus is quantified in the positive multi reaction mode (MRM) using a tandem mass spectrometer.
The assay is linear from 1 to 50 ng/ml. Inter-assay variability (3.6%-6.1%) and accuracy (91.7%-101.6%) as assessed over 20 days meet acceptance criteria. Average extraction recovery is 95.5%. There are no relevant carry-over, matrix interferences and matrix effects. Tacrolimus is stable in dried blood spots at RT and at +4 &#176;C for 1 week. Extracted samples in the autosampler are stable at +4 &#176;C for at least 72 hr.

Here we describe a high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) assay to quantify the immunosuppressant tacrolimus in dried blood spots using a simple manual protein precipitation step and online column extraction.

在免疫抑制剂他克莫司对干血斑使用LC-MS / MS定量

The calcineurin inhibitor tacrolimus is the cornerstone of most immunosuppressive treatment protocols after solid organ transplantation in the United ...

In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose

High-grade serous ovarian cancer (HGSC), the cause of widespread peritoneal metastases, continues to have an extremely poor prognosis; fewer than 30% of women are alive 5 years after diagnosis. The omentum is a preferred site of HGSC metastasis formation. Despite the clinical importance of this microenvironment, the contribution of omental adipose tissue to ovarian cancer progression remains understudied. Omental adipose is unusual in that it contains structures known as milky spots, which are comprised of B, T, and NK cells, macrophages, and progenitor cells surrounding dense nests of vasculature. Milky spots play a key role in the physiologic functions of the omentum, which are required for peritoneal homeostasis. We have shown that milky spots also promote ovarian cancer metastatic colonization of peritoneal adipose, a key step in the development of peritoneal metastases. Here we describe the approaches we developed to evaluate and quantify milky spots in peritoneal adipose and study their functional contribution to ovarian cancer cell metastatic colonization of omental tissues both in vivo and ex vivo. These approaches are generalizable to additional mouse models and cell lines, thus enabling the study of ovarian cancer metastasis formation from initial localization of cells to milky spot structures to the development of widespread peritoneal metastases.

In Vivo and Ex Vivo Approaches to Study Ovarian Cancer Metastatic Colonization of Milky Spot Structures in Peritoneal Adipose

We outline a protocol that implements both in vivo and ex vivo approaches to study ovarian cancer colonization of peritoneal adipose tissues, particularly the omentum. Furthermore, we present a protocol to quantitate and analyze immune cell-structures in the omentum known as milky spots, which promote metastases of peritoneal adipose.

在体内和体外途径研究银河系结构现货卵巢癌转移定植腹膜脂肪

High-grade serous ovarian cancer (HGSC), the cause of widespread peritoneal metastases, continues to have an extremely poor prognosis; fewer than 30% of ...

A Method of Trigonometric Modelling of Seasonal Variation Demonstrated with Multiple Sclerosis Relapse Data

This report describes a novel Stata-based application of trigonometric regression modelling to 55 years of multiple sclerosis relapse data from 46 clinical centers across 20 countries located in both hemispheres. Central to the success of this method was the strategic use of plot analysis to guide and corroborate the statistical regression modelling. Initial plot analysis was necessary for establishing realistic hypotheses regarding the presence and structural form of seasonal and latitudinal influences on relapse probability and then testing the performance of the resultant models. Trigonometric regression was then necessary to quantify these relationships, adjust for important confounders and provide a measure of certainty as to how plausible these associations were. Synchronization of graphing techniques with regression modelling permitted a systematic refinement of models until best-fit convergence was achieved, enabling novel inferences to be made regarding the independent influence of both season and latitude in predicting relapse onset timing in MS. These methods have the potential for application across other complex disease and epidemiological phenomena suspected or known to vary systematically with season and/or geographic location.

Combining plot analysis with trigonometric regression is a robust method for exploring complex, cyclical phenomena such as relapse onset timing in multiple sclerosis (MS). This method enabled unbiased characterisation of seasonal trends in relapse onset permitting novel inferences around the influence of seasonal variation, ultraviolet radiation (UVR) and latitude.

季节变化的三角建模的方法证明与多发性硬化症复发数据

This report describes a novel Stata-based application of trigonometric regression modelling to  55 years of multiple sclerosis relapse data from 46 ...

Apical Resection Mouse Model to Study Early Mammalian Heart Regeneration

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring cardiomyocytes lost due to injury and disease. In contrast to the robust cardiac regeneration of certain lower vertebrates, adult mammalian hearts typically show minimal capacity for heart regeneration and repair. However, recent studies have sparked considerable scientific interest with the finding that, between postnatal day 1 to 7 (P1 to P7), the neonatal mouse heart retains significant regenerative potential after apical resection (i.e., surgical amputation and exposure of left ventricular apex). One major controversy over this finding might be due to the diverse surgery-related procedures used in efforts to replicate or expand upon this important finding. These instructions dynamically present the materials and methodology for apical resection in a mouse model. The salient steps of this rodent survival surgery involve hypothermia anesthesia, thoracotomy, surgical amputation of heart ventricular apex, and suture and recovery of mice. The approach described could expand the application of the apical resection mouse model for cardiovascular research.

A step-by-step video protocol of apical resection is demonstrated in this study. Apical resection is a recently highlighted surgical approach in mammalian heart regeneration research. This study may promote the application of apical resection as a standard methodology in research into the mechanism underlying heart regeneration.

根尖切除小鼠模型来研究早期哺乳动物心脏再生

Cardiovascular disease plagues the whole world due to intensive lifestyle changes. Heart regeneration holds great promise for repairing and restoring ...

Assessment of Child Anthropometry in a Large Epidemiologic Study

A high proportion of children have overweight and obesity in the United States and other countries. Accurate assessment of anthropometry is essential to understand health effects of child growth and adiposity. Gold standard methods of measuring adiposity, such as dual X-ray absorptiometry (DXA), may not be feasible in large field studies. Research staff can, however, complete anthropometric measurements, such as body circumferences and skinfold measurements, using inexpensive portable equipment. In this protocol we detail how to obtain manual anthropometric measurements from children, including standing and sitting height, weight, waist circumference, hip circumference, mid-upper arm circumference, triceps skinfold thickness, and subscapular skinfold thickness, and procedures to assess the quality of these measurements. To demonstrate accuracy of these measurements, among 1,110 school-aged children in the pre-birth cohort Project Viva we calculated Spearman correlation coefficients comparing manual anthropometric measurements with a gold standard measure of body fat, DXA fat mass1. To address reliability, we evaluate intra-rater technical error of measurement at a quality control session conducted on adult female volunteers.

In epidemiologic studies of children, well-trained research staff can accurately and precisely assess weight, height, sitting height, skinfold thicknesses, and body circumferences.

大型流行病学研究儿童人体测量评估

A high proportion of children have overweight and obesity in the United States and other countries. Accurate assessment of anthropometry is essential to ...

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

The human placenta is highly inaccessible for research while still in utero. The current understanding of human placental physiology in vivo is therefore largely based on animal studies, despite the high diversity among species in placental anatomy, hemodynamics and duration of the pregnancy. The vast majority of human placenta studies are ex vivo perfusion studies or in vitro trophoblast studies. Although in vitro studies and animal models are essential, extrapolation of the results from such studies to the human placenta in vivo is uncertain. We aimed to study human placenta physiology in vivo at term, and present a detailed protocol of the method. Exploiting the intraabdominal access to the uterine vein just before the uterine incision during planned cesarean section, we collect blood samples from the incoming and outgoing vessels on the maternal and fetal sides of the placenta. When combining concentration measurements from blood samples with volume blood flow measurements, we are able to quantify placental and fetal uptake and release of any compound. Furthermore, placental tissue samples from the same mother-fetus pairs can provide measurements of transporter density and activity and other aspects of placental functions in vivo. Through this integrative use of the 4-vessel sampling method we are able to test some of the current concepts of placental nutrient transfer and metabolism in vivo, both in normal and pathological pregnancies. Furthermore, this method enables the identification of substances secreted by the placenta to the maternal circulation, which could be an important contribution to the search for biomarkers of placenta dysfunction.

The 4-vessel Sampling Approach to Integrative Studies of Human Placental Physiology In Vivo

We present a detailed method to study human placental physiology in vivo at term. The method combines blood sampling from the incoming and outgoing vessels on the maternal and fetal sides of the placenta with ultrasound measurements of volume blood flow and placental tissue sampling.

人体胎盘生理学综合研究的4血管取样方法<em&gt;体内</em

The human placenta is highly inaccessible for research while still in utero. The current understanding of human placental physiology in vivo is therefore ...