This work was supported by a Howard Hughes Medical Institute Education Grant and an internal grant from the Saint Joseph's University chapter of Sigma Xi. We would also like to thank the three anonymous reviewers who helped strengthen the protocol and interpretations.

为住房，保健和斑马鱼的研究以下方法已经批准圣约瑟夫大学IACUC。 1.斑马鱼的居所和照顾<ol><li>获取主题斑马鱼从本地供应商，还是从野外捕获的人群。请注意，住房斑马鱼是受IACUC的指导方针和额外的许可证都需要住房野生捕捞种群。 </li><li>根据性别鉴定和独立的斑马鱼。另外，请跳过此步骤或根据研究假设，分离不同的方法进行更换。 注:性别特征是难以确定在斑马鱼，尤其是在女性。参照本指引执行10或有用的在线手册来区分性别。 </li><li>随机进一步分离种群成焦鱼的适当比例对刺激鱼。注:焦鱼考试科目随后通过一系列behavio的拉尔测定，并刺激鱼是将占据侵略和浅水实验刺激室的个人。 注意:传统上，1和5之间每焦鱼刺激鱼在浅水测定法和1:1的刺激:用于在侵略测定14,17焦鱼比。然而，研究人员需要专门设计时考虑的假设研究（ 即，应该刺激鱼保持不变，保持在焦鱼行为的一致性，还是应该刺激鱼被每个检测之间旋转，以确保适当的随机化）。 </li><li>严格的12时12分光/暗周期下存储在斑马鱼房架所有的鱼（27.5-28.5℃）。日采食所有重点和激励鱼糜食品和每周一次的丰年虾，对非研究天。 </li><li>确保所有测试ZT1和ZT5（时间授时）之间进行，以避免昼夜节律的影响26。 </li></ol> 2.随机化和坦克安装<ol><li>镭ndomize所有个人先验行为检测的特定序列。设置使用在线随机序列发生器27或从统计软件包（R）的一个随机序列发生器一致随机化方案。 <ol><li>标签1每个检测到3，并保持这个任务是一致的。分配一个随机序列将每个独立的，并根据行为分析标签映射序列。例如，对于总共三个试验（1映射到变浅，2映射到胆识和3映射到侵略），一个给定的个体可以具有&#39;2,3,1&#39;的一个随机序列，这将要求它们执行特定行为检测指定的顺序排列。 </li></ol></li><li>设立在实验室试验罐（胆识，侵略和浅水）在尽量减少外界不良刺激的可能性，并确保有足够的空间用于视频摄像机捕捉整个坦克的地方。确保摄像机已enoug^ h清晰查看罐的标签，以及能够区分焦点鱼和安装在这个位置的摄像机的微妙行为。 </li><li>记录与保持在与指定的箱的全视图，以尽量减少在行为得分作为差分录音的结果意外差别相同位置的相机中的所有行为试验。 注意:避免将坦克重型交通区域和门的动作和声音会影响斑马鱼的行为。 </li><li>建立独特的JWatcher协议24得分每个行为的检测。请参阅JWatcher手册，了解如何建立联络主文件分配行为，以键盘和焦点分析​​文件，总结收集到的数据。 注:JWatcher是行为记分软件，允许可靠和可重复的定制行为得分24。 </li><li>定义标记方案来跟踪日期，鱼数量，并为所有的行为实验测定编号（ 如:"F5-B-123115"暗示鱼5，胆识检测，12月31日日 ，2015年），并在标签视图和摄像机的焦点罐用胶带之外。保持并记录在实验室笔记本为快速参考和未来的质量控制信息。 </li><li>设置将所有测定之间使用以及直接在个人从壳体齿条移除之后的驯化槽。 注:驯化槽和终端舱是小的水族馆砾石英寸（40厘米×20厘米×25厘米），并且接近于壳体机架的过滤器。 </li><li>该实验全部完成后，转移个人到一个单独的终端坦克将作为所有使用焦距个人最终目的地坦克。使用槽是大到足以容纳一组个体的生命（ 例如25加仑）的其余部分。 注:保持一致和相似的水的温度和条件ACROS这是非常重要的一切都坦克。这包括住房，驯化，和所有的测试坦克。 </li></ol> 3.开展侵略分析<ol><li>进行侵略测定在一个19升矩形槽测量30厘米×15厘米×10厘米。装在围绕侵略坦克三面外不透明隔板槽，留，以观看前面，以减少无用的外界刺激。分与外部标记罐分成四个相等的矩形象限并永久固定的反射镜（用硅酮填缝）左下象限内，在22.5°倾斜形成对罐的最左侧一个直角三角形。 </li><li>确保水在驯化槽和侵略罐中的温度在壳体机架2℃。要做到这一点，有新鲜的，老化水预热至27.5-28.5℃，准备加入到侵略坦克。请注意，以改变水在每个测定之间，以消除无端是重要嗅觉线索。 </li><li>转移，通过拔罐，焦点鱼从壳体机架到一个单独的驯化槽那里将适应环境10分钟。 注:重要的是要通过降低处理不当引入到鱼的刺激。传输个人如前所述27。 <ol><li>具体地，手动和小心地将一个小的，透明的塑料杯到感兴趣罐。而不破坏鱼不亚于通过使用净转移，慢慢鱼舀进杯子。 </li></ol></li><li>确保视频摄像机准备通过确认试验侵略罐（见步骤3.1）是焦点，并立即个体转移到侵略罐，在设备上按记录之前记录。通过拔罐从顺应坦克侵略罐转移焦点鱼。 注:记录鱼进入坦克侵略跟踪条目的确切时间之前是很重要的。 </li><li>记录汇聚分裂国家罐10分钟，30秒，然后杯从侵略罐放回驯化槽的焦点的鱼。 </li><li>在观看录制，等待30秒的鱼已经被引入到罐后的进球行为之前，一个额外的适应期。 </li><li>使用用于攻击行为限定的JWatcher评分系统，量化为10分钟以下行为:邻近镜花费时间，镜像方法，飞镖的数量，试图叮咬（在反射镜的方向张口）的数目，和横向显示的号码数（背鳍，胸鳍，臀鳍和尾鳍勃起）9。 </li><li>分数单侵略试验至少两次，直到检测与迭代之间的一致性高得分。 </li></ol> 4.开展大胆试验<ol><li>在开展小1.5升梯形水箱大胆试验，​​测量15厘米高x26.5厘米顶的X22.5厘米底×6厘米宽。环绕四面不透明的纸ØF中的坦克，但留下暴露观察前方。一个单一的，薄片胶带外部的放置在罐从罐的底部划界的顶部。 </li><li>确保水在大胆罐中的温度在壳体机架2℃（参见步骤3.2） </li><li>从壳体机架驯化槽转移焦点鱼（参见步骤3.3），并让适应环境10分钟。 </li><li>确保摄像机准备拍摄大胆试验，​​一旦准备转移焦点鱼，按记录设备。从驯化罐转移焦点鱼的胆量槽和录制了8分30秒的。 </li><li>当观看记录，允许另外的30秒的适应期的鱼被引入到得分前油箱之后。 </li><li>使用JWatcher软件，量化大胆行为8分钟。具体来说，量化以下行为:近表面的时间，延迟进入TRA的上部，数nsitions进入上部，和29不稳定的运动（飞镖）的数目。 </li><li>分数单大胆试验至少两次，直到检测与迭代之间的一致性高得分。 </li></ol> 5.开展了浅水分析<ol><li>进行浅水实验中有三个隔间用硅胶密封填缝两个玻璃隔板分成一个76升罐（所产生的三个室由两个刺激区和一个联络室）。在外部，标记指定每个刺激区&#39;玻璃分隔的6.35厘米双重偏好的区域，表明两种车身长度30。应用不透明的分区外部周围的浅水坦克的三面，但留下暴露观察前方。 注:此首选项区可进行定制，以测试不同大小的物种的特定假设。 </li><li>根据研究的性质，选择刺激滩涂适应环境至少12小时beforÈ测试并保留在浅水罐在整个实验过程中。 注:刺激滩涂是用来评估焦点鱼在这些浅滩的存在或不存在的行为鱼个体的集合。不同的刺激浅滩组合可以选择测试多个不同的浅水假设。可以肯定的刺激浅滩各自刺激区域之间旋转，以避免侧偏置，并且也使其适应测试前至少12小时。 </li><li>如前所述转移焦点鱼入驯化罐（见步骤3.3） </li><li>传送，通过从中间驯化槽在浅水罐中央室拔罐，焦点鱼。 </li><li>允许10分钟的现场得分的观察期前的10分钟适应期。另外，记录浅水试验，然后再使用JWatcher得分。鱼被确定为变浅时，他们在腔室中的优先区域内住房目标浅滩。 </li><li>量化在中心容积花各人喜好区和时间花费的时间。 </li><li>作为额外的描述计量，监测"浅水的实力"（SOS）。其中大小描述如何经常单个鱼群分配-1和+1之间的值。 </li><li>计算由浅水区的面积除以所花费的时间变浅的比率和减去的时间的比例花费不变浅和非浅水区的面积。放置此数量超过由每个区域的体积单独分开的每个区所花费的时间的总和。关于该计算的详细的信息可以在先前的研究中14中找到。 </li></ol> 6，数据质量控制<ol><li>分析使用传统的数据分析工具31-32的数据。 </li><li>测试使用非参数列文的测试的所有行为方差齐性。如果一个行为是不均匀的执行日志变换33。 </li><li>确认没有顺序的影响（ 即，随机化为了不影响个人的行为）对同质数据34使用秩和检验方差分析。 </li><li>确认有与侧固定系数35 SOS的指数创建单变量一般线性模型无副作用偏见的浅水检测。 注意:如果有一点与数据没有问题，下游分析可以进行。如果有问题，调和额外的转换，具有生物合理的解释合理，和/或增加样本量。 </li></ol> 7.分析数据<ol><li>计算双尾Spearman秩相关性36分开内的胆略和侵略检测所有行为。 </li><li>计算组内相关系数（ICC），为与行为分析稠度进行了一个以上的添双向混合模型E对于一个给定的个人36。在这一背景下，国际刑事法院是在给定的实验乖鱼如何衡量持续超过一次的测量。 </li><li>由中心和拉平所有行为预处理魄力和侵略行为，以输入准备一个主成分分析（PCA）25。 注:由中心和拉平预处理测量是一个重要的步骤。如果没有可变更多的次数多，会出现过度影响提取成分的解释这个预处理步骤的行为。居中和拉平的过程中会删除此偏差。 </li><li>执行与相关矩阵PCA一，以确定是否魄力和侵略是作为行为综合症联系在了一起。在&gt; 0.6诠释每个组件显著的行为负荷。见2010 Budaev 24报告PCA的行为学研究的综合指南。一个碎石图将知识性上eigenve构建函数应保持解释。 </li><li>使用统计软件包如R或SPSS 31,32，提取得分为每个单独的每个组件。该分数代表多少给定的个人的行为可以由特定的组件进行说明。 </li><li>计算因素回归分数和不冻36的强度之间的Spearman等级相关。 注:行为综合症被视为存在，如果魄力和侵略行为加载相同的组件上，或者，如果有显著可解释的负载任何组件与浅水的实力相关。另外，还要注意浅水的实力不包含在PCA，以允许更多复杂的行为quantifications（胆略和侵略）更好的解释。该PCA将正交崩溃魄力和侵略特性引入特征向量，最大限度的变化说明，并非常适用于这些复杂的测量量。解读所产生的特征向量和回归我们的浅水测量强度对他们每个人给出了每个复杂的行为（胆略和侵略）涉及到一个更明确的二进制浅水测量一个更加直观的解释（ 即鱼是不冻或不不冻）。 </li></ol>

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The authors (GW and SPM) have no competing financial interests or conflicts of interest.

该协议将确定是否有在魄力，侵略一致协会，和在斑马鱼变浅的行为。如果有一致的关联的任何这些行为之间的给定人群中，然后一个行为综合症存在。通过研究人口的自然行为综合症，研究人员可以有它的行为的动态的一个更完整的理解，种群结构，以及可能的进化史3。此外，操纵影响这些行为综合征，像在药理学6，毒理学7，行为遗传学8,9的环境中，和内分?...

有文献的从一个给定的群体调查个别动物内不同的行为之间的关联越来越多。这些协会被称为行为综合征，并测量通常包括气魄，侵略，探索行为和社交1-5。行为综​​合征是直接和间接原因有价值。直接，行为症状的知识，可以提供进化理论，人口结构和人口动态3的更完整视图。间接的，对于行为的关联知识可能通知量化的行为，如药理6，7毒理学，行为遗传学8,9，和内分泌10个字段。因为这些直接和间接的利益，行为症状的知识的增加是常用的模式生物，如斑马鱼特别有价值。使用斑马鱼的研究发现我呐多种学科，包括行为综合征11-13的分析。为了推进行为综合症的研究知识，因为其他学科也使用行为三围为假设检验，性能可靠，简洁的描述都需要有效的分析和解释，标准化的协议将有利于物种的学习间的比较。我们的协议被开发用于测量实验室饲养的斑马鱼14人口的气魄侵略，浅水行为综合症。然而，该协议的基础上（跟踪单个鱼，确保适当的随机化，以及适当的分析）可以很容易地修改各种替代行为的措施。此外，气魄，侵略，或浅水测定可单独针对不同的假设进行测试运行。因此，尽管我们的目标是描述如何进行行为综合症的研究和成功的人，微升协议埃维尔行为测量，此过程的每个方面都可以单独存在。 对行为综合征的文献跨越几个分类组，从节肢动物对人类4和，以测量一个行为综合症，至少两个行为背景必须量化。不幸的是，在用于量化横跨行为的轴线的行为测量测定法往往很少的一致性。例如，在鱼，粗体可能使用T-迷宫试验，旷场试验中，或引入一种新的或外国的刺激15的测量。在鱼的侵略研究可能涉及对子互动，视频刺激试验，或粘土模型试验12,16,17。同样，变浅的行为，这通常涉及shoalmate偏爱的测量分析，可以在不同类型的罐进行的，用不同的方法来确定缔合时间21-23。在这个协议中的overa的特定子集LL行为分析的剧目呈现。具体而言，该协议提出了一个方法来跟踪通过气魄，侵略和浅水测定个人，有利于个体内比较，以确定比较是否在整个人口中的所有个体一致的方式。我们在以前的研究中14,18执行该协议与斑马鱼和慈鲷罪犯（Amatitlania nigrofasciata），它可以与任何同样大小的淡水鱼工作。 魄力测定法中，有一个水平行划定在水箱的上部和下部相等大小的区域一个1.5升梯形罐进行。量化行为包括跃迁受水箱的上部和下部，在各部分，飞镖的数量所花费的时间之间的受试鱼的数目，并进入上部的延迟。侵略测定在一个19L的矩形箱，其包括执行一个3英寸×5英寸的镜面倾斜约22°，位于罐19的左下角。量化行为包括由目标鱼与镜20相互作用花费的总时间量，具有特定侵略性指标沿-试验鱼和其反射之间叮咬的数目，横向显示，飞镖。对于这些具体指标，叮咬定义为走向张开嘴镜子快速弓步，横向显示被定义为横向，胸鳍，臀鳍和背鳍的在镜子的方向燃烧，和飞镖任何不稳定的运动，是不朝镜子执导。最后，变浅测定量化测试鱼类中的三腔槽的中心腔室中的行为。该储箱的侧室要么空，或者含有鱼的"目标浅滩"，并且测试鱼花费每侧室接近的时间被测量21-23。单个综合得分称为STREN变浅的GTH（SOS），计算每个单独的测试鱼类，具体到刺激，并且可以在下游分析14一起使用。所有行为是由一个单一的浏览器，或使用被称为JWatcher 24自由行为量化软件多个观看者得分。 测试行为综合症的存在主要是一个统计的努力，并且在2010年Budeav所提出，建议遵循的准则25，具体而言，建议在中心和范数据执行主分量分析（PCA），其中输入是一个人的行为与多种行为测量 （ 即气魄和侵略）测定的载体。在PCA上的相关矩阵进行的，减少了行为测量的维数，因此，提取，解释大部分变化的最重要的知识。然后将提取的部件可以被解释浅上高的因子载荷为感兴趣和回归分数个体行为编辑可以提取用于说明组件的基础上，每一个人。然后，这些回归的分数可以比在SOS测量等各种非行为的测量，如鱼的大小或性别。 这个工作流程是在该胆识和浅水14之间存在特定的性别行为综合征被发现斑马鱼的行为综合征的研究中得到落实。在这种情况下，更大胆的斑马鱼男性更可能具有较大的，更积极的物种（D. albolineatus）相关联，但是这种关联在雌性丢失。该工作流还在其一个行为综合症未发现的，潜在地表明该物种的行为可塑性少年囚犯鲷亲属（Amatitlania nigrofasciata）18的研究来实现。因此，下面的协议，提出瓦特第i个在研究个人层面的行为综合症的框架划定的三个具体分析（气魄，侵略和浅水）性质的目标。

<table border="0" cellpadding="0" cellspacing="0" width="783">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:352px;">Zebrafish Rack System</td>
			<td style="width:224px;">Aquaneering Inc</td>
			<td style="width:119px;">Cat. # ZS550</td>
			<td style="width:88px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pet Valu Tropical Fish Food, 224.0g</td>
			<td>Pet Valu</td>
			<td>Cat. # 31700</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Premium Grade Brine Shrimp Eggs, 16 oz</td>
			<td>Brine Shrimp Direct</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">1.5 L Trapezoidal Tank</td>
			<td>Pentair Aquatic Ecosystems</td>
			<td>Cat. # itsts-a</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">19L rectangular tank</td>
			<td>That Fish Place</td>
			<td align="right">211932</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">76L rectangular tank</td>
			<td>That Fish Place</td>
			<td align="right">212180</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hitachi KP-D20A CCD Camera</td>
			<td>Prescott&#39;s, Inc.</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Nikon AF Nikkor 35-105mm f/305~4.5s MACRO lens</td>
			<td>Nikon Corporation</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">ArtMinds Square Mirror, Value Pack 3&#34;x3&#34;</td>
			<td>Michaels</td>
			<td>Cat. # 10334162</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Jwatcher</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">SPSS Statistics Base</td>
			<td>IBM</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">R</td>
			<td>The R Foundation</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

boldness, aggression, and shoaling assays for zebrafish behavioral syndromes

在具体的行为在不同的上下文交互一种行为综合征的存在。斑马鱼已经测试对象中最近的研究和标准化的协议，以确保正确的分析和解释是重要的。在我们以前的研究中，我们通过监测了一系列的1.5升水箱的梯形行为（近地表的时间，潜伏期过渡，过渡的数量和飞镖）的测量气魄。同样，我们在一个长方形19升油箱量化侵​​略观察叮咬，横向显示，飞镖和时间近一个倾斜的镜子。通过将一个76升罐投入三分，我们还研究了浅水偏好。变浅检测是一个高度可定制的试验和可以为特定的假设进行调整。然而，对于该测定的协议也必须是标准化的，但是定制不够灵活。在以前的研究中，末端腔要么空的，包含5或10的斑马鱼，或5闪电（鱼丹）（D. albolineatus）。在FOL哞哞叫的手稿，我们提出了一个详细的协议，并伴随该协议，该协议将允许的行为综合症实验复制的成功应用的代表性数据。

根据研究的性质，和所用的具体协议，几种不同的结果在一个行为综合征实验是可能的。下面的表格和数字，其中指出，改编自我们以前的杂志上发表的研究行为流程14和斑马鱼杂志17。当提案（如上所述）将其整体，两组结果，"测定法相关&#39;内，并进行"测定法相关之间，"预期（ 图1）。 所述第...

Watch this Scientific Journal Video about Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes at JoVE.com

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

气魄，侵略和浅水测定斑马鱼行为综合征

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

boldness-aggression-shoaling-assays-for-zebrafish-behavioral

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10.2K 次观看.  Saint Joseph’s University. This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

视频: Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

Intra-Operative Behavioral Tasks in Awake Humans Undergoing Deep Brain Stimulation Surgery

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through implanted electrodes. Deep brain stimulation was first implemented as a therapeutic modality by Benabid et al. in the late 1980s, when he used this technique to stimulate the ventral intermediate nucleus of the thalamus for the treatment of tremor 1. Currently, the procedure is used to treat patients who fail to respond adequately to medical management for diseases such as Parkinson's, dystonia, and essential tremor. The efficacy of this procedure for the treatment of Parkinson's disease has been demonstrated in well-powered, randomized controlled trials 2. Presently, the U.S. Food and Drug Administration has approved DBS as a treatment for patients with medically refractory essential tremor, Parkinson's disease, and dystonia. Additionally, DBS is currently being evaluated for the treatment of other psychiatric and neurological disorders, such as obsessive compulsive disorder, major depressive disorder, and epilepsy. 
DBS has not only been shown to help people by improving their quality of life, it also provides researchers with the unique opportunity to study and understand the human brain. Microelectrode recordings are routinely performed during DBS surgery in order to enhance the precision of anatomical targeting. Firing patterns of individual neurons can therefore be recorded while the subject performs a behavioral task. Early studies using these data focused on descriptive aspects, including firing and burst rates, and frequency modulation 3. More recent studies have focused on cognitive aspects of behavior in relation to neuronal activity 4,5. This article will provide a description of the intra-operative methods used to perform behavioral tasks and record neuronal data with awake patients during DBS cases. Our exposition of the process of acquiring electrophysiological data will illuminate the current scope and limitations of intra-operative human experiments.

Deep brain stimulation surgery offers a unique opportunity to examine information encoding in the awake human brain. This article will describe intra-operative methods used to perform cognitive and behavioral tasks while simultaneously acquiring physiological data such as EMG, single-unit neuronal activity and/or local field potentials.

在接受深部脑刺激手术术中唤醒人类行为任务

Deep brain stimulation (DBS) is a surgical procedure that directs chronic, high frequency electrical stimulation to specific targets in the brain through ...

科研

医学

Using Visual and Narrative Methods to Achieve Fair Process in Clinical Care

The Institute of Medicine has targeted patient-centeredness as an important area of quality improvement. A major dimension of patient-centeredness is respect for patient's values, preferences, and expressed needs. Yet specific approaches to gaining this understanding and translating it to quality care in the clinical setting are lacking. From a patient perspective quality is not a simple concept but is best understood in terms of five dimensions: technical outcomes; decision-making efficiency; amenities and convenience; information and emotional support; and overall patient satisfaction. Failure to consider quality from this five-pronged perspective results in a focus on medical outcomes, without considering the processes central to quality from the patient's perspective and vital to achieving good outcomes. In this paper, we argue for applying the concept of fair process in clinical settings. Fair process involves using a collaborative approach to exploring diagnostic issues and treatments with patients, explaining the rationale for decisions, setting expectations about roles and responsibilities, and implementing a core plan and ongoing evaluation. Fair process opens the door to bringing patient expertise into the clinical setting and the work of developing health care goals and strategies. This paper provides a step by step illustration of an innovative visual approach, called photovoice or photo-elicitation, to achieve fair process in clinical work with acquired brain injury survivors and others living with chronic health conditions. Applying this visual tool and methodology in the clinical setting will enhance patient-provider communication; engage patients as partners in identifying challenges, strengths, goals, and strategies; and support evaluation of progress over time. Asking patients to bring visuals of their lives into the clinical interaction can help to illuminate gaps in clinical knowledge, forge better therapeutic relationships with patients living with chronic conditions such as brain injury, and identify patient-centered goals and possibilities for healing. The process illustrated here can be used by clinicians, (primary care physicians, rehabilitation therapists, neurologists, neuropsychologists, psychologists, and others) working with people living with chronic conditions such as acquired brain injury, mental illness, physical disabilities, HIV/AIDS, substance abuse, or post-traumatic stress, and by leaders of support groups for the types of patients described above and their family members or caregivers.

This paper illustrates an innovative visual approach (photovoice or photo-elicitation) to achieve fair process in clinical care for patients living with chronic health conditions, illuminate gaps in clinical knowledge, forge better therapeutic relationships, and identify patient-centered goals and possibilities for healing.

利用视觉和叙事方法，以实现公平的进程中，在临床护理

The Institute of Medicine has targeted patient-centeredness as an important area of quality improvement. A major dimension of patient-centeredness is ...

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 ...

Eye Tracking Young Children with Autism

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a direct, detailed and objective measure of gaze behavior, eye-tracking has become a valuable tool for examining abnormal perceptual strategies in clinical populations and has been used to identify disorder-specific characteristics1, promote early identification2, and inform treatment3. In particular, investigators of autism spectrum disorders (ASD) have benefited from integrating eye-tracking into their research paradigms4-7. Eye-tracking has largely been used in these studies to reveal mechanisms underlying impaired task performance8 and abnormal brain functioning9, particularly during the processing of social information1,10-11. While older children and adults with ASD comprise the preponderance of research in this area, eye-tracking may be especially useful for studying young children with the disorder as it offers a non-invasive tool for assessing and quantifying early-emerging developmental abnormalities2,12-13. Implementing eye-tracking with young children with ASD, however, is associated with a number of unique challenges, including issues with compliant behavior resulting from specific task demands and disorder-related psychosocial considerations. In this protocol, we detail methodological considerations for optimizing research design, data acquisition and psychometric analysis while eye-tracking young children with ASD. The provided recommendations are also designed to be more broadly applicable for eye-tracking children with other developmental disabilities. By offering guidelines for best practices in these areas based upon lessons derived from our own work, we hope to help other investigators make sound research design and analysis choices while avoiding common pitfalls that can compromise data acquisition while eye-tracking young children with ASD or other developmental difficulties.

Eye tracking has long been used to study gaze patterns in typically-developing individuals, but recent technological advancements have made its use with clinical populations, including autism, more feasible. While eye-tracking young children with autism can offer insight into early symptom manifestations, it involves methodological challenges. Suggestions for best practices are provided.

自闭症幼儿的眼动追踪

The rise of accessible commercial eye-tracking systems has fueled a rapid increase in their use in psychological and psychiatric research. By providing a ...

Processing of Primary Brain Tumor Tissue for Stem Cell Assays and Flow Sorting

Brain tumors are typically comprised of morphologically diverse cells that express a variety of neural lineage markers. Only a relatively small fraction of cells in the tumor with stem cell properties, termed brain tumor initiating cells (BTICs), possess an ability to differentiate along multiple lineages, self-renew, and initiate tumors in vivo. We applied culture conditions originally used for normal neural stem cells (NSCs) to a variety of human brain tumors and found that this culture method specifically selects for stem-like populations. Serum-free medium (NSC) allows for the maintenance of an undifferentiated stem cell state, and the addition of bFGF and EGF allows for the proliferation of multi-potent, self-renewing, and expandable tumorspheres.
To further characterize each tumor's BTIC population, we evaluate cell surface markers by flow cytometry. We may also sort populations of interest for more specific characterization. Self-renewal assays are performed on single BTICs sorted into 96 well plates; the formation of tumorspheres following incubation at 37 &deg;C indicates the presence of a stem or progenitor cell. Multiple cell numbers of a particular population can also be sorted in different wells for limiting dilution analysis, to analyze self-renewal capacity. We can also study differential gene expression within a particular cell population by using single cell RT-PCR.
The following protocols describe our procedures for the dissociation and culturing of primary human samples to enrich for BTIC populations, as well as the dissociation of tumorspheres. Also included are protocols for staining for flow cytometry analysis or sorting, self-renewal assays, and single cell RT-PCR.

The identification of brain tumor initiating cells (BTICs), the rare cells within a heterogeneous tumor possessing stem cell properties, provides new insights into human brain tumor pathogenesis. We have refined specific culture conditions to enrich for BTICs, and we routinely use flow cytometry to further enrich these populations. Self-renewal assays and transcript analysis by single cell RT-PCR can subsequently be performed on these isolated cells.

加工的主要脑肿瘤组织的干细胞检测和流量排序

Brain tumors are typically comprised of morphologically diverse cells that express a variety of neural lineage markers. Only a relatively small fraction ...

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Genomic studies of human cancers have yielded a wealth of information about genes that are altered in tumors1,2,3. A challenge arising from these studies is that many genes are altered, and it can be difficult to distinguish genetic alterations that drove tumorigenesis from that those arose incidentally during transformation. To draw this distinction it is beneficial to have an assay that can quantitatively measure the effect of an altered gene on tumor initiation and other processes that enable tumors to persist and disseminate. Here we present a rapid means to screen large numbers of candidate melanoma modifiers in zebrafish using an autochthonous tumor model4 that encompasses steps required for melanoma initiation and maintenance. A key reagent in this assay is the miniCoopR vector, which couples a wild-type copy of the mitfa melanocyte specification factor to a Gateway recombination cassette into which candidate melanoma genes can be recombined5. The miniCoopR vector has a mitfa rescuing minigene which contains the promoter, open reading frame and 3'-untranslated region of the wild-type mitfa gene. It allows us to make constructs using full-length open reading frames of candidate melanoma modifiers. These individual clones can then be injected into single cell Tg(mitfa:BRAFV600E);p53(lf);mitfa(lf)zebrafish embryos. The miniCoopR vector gets integrated by Tol2-mediated transgenesis6 and rescues melanocytes. Because they are physically coupled to the mitfa rescuing minigene, candidate genes are expressed in rescued melanocytes, some of which will transform and develop into tumors. The effect of a candidate gene on melanoma initiation and melanoma cell properties can be measured using melanoma-free survival curves, invasion assays, antibody staining and transplantation assays.

A rapid way to screen for melanoma modifiers using a zebrafish autochthonous tumor model is presented. It takes advantage of the miniCoopR vector which allows for expression of candidate melanoma genes in melanocytes. A method to obtain melanoma-free survival curves, an invasion assay, a protocol for antibody staining of scale melanocytes and a melanoma transplantation assay are described.

对黑色素瘤修饰符一个斑马鱼原地肿瘤模型的筛选

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

Adaptation of Semiautomated Circulating Tumor Cell (CTC) Assays for Clinical and Preclinical Research Applications

The majority of cancer-related deaths occur subsequent to the development of metastatic disease. This highly lethal disease stage is associated with the presence of circulating tumor cells (CTCs). These rare cells have been demonstrated to be of clinical significance in metastatic breast, prostate, and colorectal cancers. The current gold standard in clinical CTC detection and enumeration is the FDA-cleared CellSearch system (CSS). This manuscript outlines the standard protocol utilized by this platform as well as two&#160;additional adapted protocols that describe the detailed process of user-defined marker optimization for protein characterization of patient CTCs and a comparable protocol for CTC capture in very low volumes of blood, using standard CSS reagents, for studying in vivo preclinical mouse models of metastasis. In addition, differences in CTC quality between healthy donor blood spiked with cells from tissue culture versus patient blood samples are highlighted. Finally, several commonly discrepant items that can lead to CTC misclassification errors are outlined. Taken together, these protocols will provide a useful resource for users of this platform interested in preclinical and clinical research pertaining to metastasis and CTCs.

Adaptation of Semiautomated Circulating Tumor Cell CTC Assays for Clinical and Preclinical Research Applications

Circulating tumor cells (CTCs) are prognostic in several metastatic cancers. This manuscript describes the gold standard CellSearch system (CSS) CTC enumeration platform and highlights common misclassification errors. In addition, two&#160;adapted protocols are described for user-defined marker characterization of CTCs and CTC enumeration in preclinical mouse models of metastasis using this technology.

半自动适应循环肿瘤细胞（CTC）分析临床和临床前研究中的应用

The majority of cancer-related deaths occur subsequent to the development of metastatic disease. This highly lethal disease stage is associated with the ...

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone resorption. Osteoclasts can be generated in vitro from monocyte/macrophage precursor cells in the presence of certain cytokines, which promote survival and differentiation. Here, both in vivo and in vitro techniques are demonstrated, which allow scientists to study different cytokine contributions towards osteoclast differentiation, signaling, and activation. The minicircle DNA delivery gene transfer system provides an alternative method to establish an osteoporosis-related model is particularly useful to study the efficacy of various pharmacological inhibitors in vivo. Similarly, in vitro culturing protocols for producing osteoclasts from human precursor cells in the presence of specific cytokines enables scientists to study osteoclastogenesis in human cells for translational applications. Combined, these techniques have the potential to accelerate drug discovery efforts for osteoclast-specific targeted therapeutics, which may benefit millions of osteoporosis and arthritis patients worldwide.

A Novel in vivo Gene Transfer Technique and in vitro Cell Based Assays for the Study of Bone Loss in Musculoskeletal Disorders

Differentiation of precursor cells into osteoclasts is regulated by cytokines and growth factors. Here, a novel gene transfer technique for differentiation of osteoclasts in vivo and cell culture protocols for differentiating precursor cells into osteoclasts in vitro as a method to study the effects of cytokines on osteoclastogenesis are described.

一种新型的<em&gt;在体内</em&gt;基因转移技术和<em&gt;在体外</em&gt;细胞分析为肌肉骨骼疾病骨质流失的研究

Differentiation and activation of osteoclasts play a key role in the development of musculoskeletal diseases as these cells are primarily involved in bone ...

A Novel Model of Mild Traumatic Brain Injury for Juvenile Rats

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor vehicle accidents, and falls, a reliable animal model of mTBI had previously not been developed for this important aspect of development. The modified weight-drop technique employs a glancing impact to the head of a freely moving rodent transmitting acceleration, deceleration, and rotational forces upon the brain. When applied to juvenile rats, this modified weight-drop technique induced clinically relevant behavioural outcomes that were representative of post-concussion symptomology. The technique is a rapidly applied procedure with an extremely low mortality rate, rendering it ideal for high-throughput studies of therapeutics. In addition, because the procedure involves a mild injury to a closed head, it can easily be used for studies of repetitive brain injury. Owing to the simplistic nature of this technique, and the clinically relevant biomechanics of the injury pathophysiology, the modified weight-drop technique provides researchers with a reliable model of mTBI that can be used in a wide variety of behavioural, molecular, and genetic studies.

The modified weight-drop technique is an easy, cost-effective procedure used for the induction of mild traumatic brain injury in juvenile rats. This novel technique produces clinically relevant symptomology that will advance the study of mild traumatic brain injury (mTBI) and concussion.

轻度脑外伤的幼年大鼠一种新模式

Despite growing evidence that childhood represents a major risk period for mild traumatic brain injury (mTBI) from sports-related concussions, motor ...

Real-time Cytotoxicity Assays in Human Whole Blood

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with high-throughput cell positioning technology. The targeted tumor cell populations are first preprogrammed to immobilization into an array format, and labeled with green fluorescent cytosolic dyes. Following the cell array formation, antibody drugs are added in combination with human whole blood. Propidium iodide (PI) is then added to assess cell death. The cell array is analyzed with an automatic imaging system. While cytosolic dye labels the targeted tumor cell populations, PI labels the dead tumor cell populations. Thus, the percentage of target cancer cell killing can be quantified by calculating the number of surviving targeted cells to the number of dead targeted cells. With this method, researchers are able to access time-dependent and dose-dependent cell cytotoxicity information. Remarkably, no hazardous radiochemicals are used. The WCA presented here has been tested with lymphoma, leukemia, and solid tumor cell lines. Therefore, WCA allows researchers to assess drug efficacy in a highly relevant ex&#160;vivo condition.

The whole blood cytotoxicity assay (WCA) is a cytotoxicity assay developed by incorporating high-throughput cell positioning technology with fluorescence microscopy and automated image processing. Here, we describe how lymphoma cells treated with an anti-CD20 antibody can be analyzed real-time in human whole blood to provide quantitative cellular cytotoxicity analysis.

实时的细胞毒性分析在人全血

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...