Name: systems analysis of the neuroinflammatory and hemodynamic response to traumatic brain injury
Uploaded: 2022-05-27T13:15:00
Description: Watch this Scientific Journal Video about Systems Analysis of the Neuroinflammatory and Hemodynamic Response to Traumatic Brain Injury at JoVE.com

该项目得到了美国国立卫生研究院R21 NS104801（EMB）和R01 NS115994（LBW / EB）以及亚特兰大儿童保健初级教师重点奖（EMB）的支持。这项工作还得到了美国国防部通过国会指导的医学研究计划的支持，奖励号为。W81XWH-18-1-0669 （LBW/EMB）.意见，解释，结论和建议是作者的意见，解释，结论和建议不一定得到国防部的认可。本材料基于美国国家科学基金会研究生研究奖学金计划支持的工作，授予编号1937971。本材料中表达的任何意见，发现，结论或建议均为作者的观点，并不一定反映美国国家科学基金会的观点。

所有动物程序均由埃默里大学机构动物护理和使用委员会（IACUC）批准，并遵循NIH实验室动物护理和使用指南。
1. 轻度创伤性脑损伤的降重模型
<ol><li>准备举重设置。将虎钳安装在平坦的表面上，并垂直对齐1米的导流管（内径2.54厘米）（使用水平仪检查）。使用54克螺栓（基本体径0.95厘米，头部直径2厘米，长度10.2厘米）进行冲击。</li>
	<li>短暂麻醉小鼠。用4.5%异氟醚在100...

<ol>
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在这里，我们详细介绍了评估对重复性轻度创伤性脑损伤的血流动力学和神经炎症反应的方法。此外，我们已经展示了如何使用偏最小二乘回归将这些数据作为多元系统分析的一部分进行积分。在下面的文本中，我们将讨论与协议相关的一些关键步骤和限制，以及这些方法相对于现有方法的优缺点。
轻度创伤性脑损伤的降重模型。 这种创伤性脑损伤诱导方法是?...

概述 轻度创伤性脑损伤（mTBIs）每年影响约160-380万运动员1.这些损伤，包括亚脑震荡和脑震荡损伤，可使患者出现短暂的躯体、情绪、心理和认知症状2.此外，在“脆弱性窗口”内持续的 重复 性mTBI（rmTBI）可能导致认知后果的累积严重程度和持续时间，其持续时间比单个mTBI单独3的影响更长，最终甚至导致功能永久丧失4，5，6。虽然许多患者在相对较短的时间范围内（<1周）恢复，但10-40%的患者在1个月内遭受>mTBI的更持久影响，有些持续长达1年3，7，8，9。尽管这些损伤的患病率很高且后果持久，但对损伤机制知之甚少，并且没有有效的治疗策略。
鉴于mTBI/rmTBI后结局的高度变异性，在确定在终末mTBI/rmTBI研究中获得的组织的早期分子触发因素的一个挑战是缺乏纵向数据证明这些分子触发因素与长期结局的明确“急性分子联系”。为了克服这一挑战，我们小组发现，使用称为弥漫相关光谱（DCS）的光学工具急性测量的急性脑血流量减少与rmTBI10小鼠模型中的长期认知结果密切相关。使用这种血流动力学生物标志物，我们表明脑血流量急性降低（并且通过扩展，预测的长期结果更差）的小鼠在MAPK和NFκB通路内同时具有神经元磷酸信号传导的急性增加，促炎细胞因子的神经元表达增加，吞噬细胞/小胶质细胞标志物的表达增加111.这些数据表明，神经元磷酸化信号传导、细胞因子表达和小胶质细胞活化在损伤后脑血流的急性调节以及触发导致神经元功能障碍和更差认知结果的信号级联反应中可能起作用。在本文中，我们详细介绍了在rmTBI之后同时探测血流动力学和神经炎症环境的方法，以及如何整合这些复杂的数据集。具体而言，我们概述了这种综合方法的四个关键步骤的程序：（1）轻度创伤性脑损伤的重量下降模型，（2）使用弥漫相关光谱评估脑血流，（3）神经炎症环境的定量，以及（4）数据整合（图1）。下面，我们简要介绍了这些关键步骤中的每一个，以帮助指导读者了解我们方法背后的基本原理。手稿的其余部分为每个关键步骤提供了详细的协议。
轻度创伤性脑损伤的降重模型 虽然存在许多重复性轻度TBI的优秀临床前模型12，13，14，15，16，17，18，但我们采用了一个完善且临床相关的重量下降闭合性头部损伤模型。该模型的主要特征包括（1）完整颅骨/头皮的钝性影响，然后头部围绕颈部的无限制旋转，（2）没有明显的结构性脑损伤，水肿，血脑屏障损伤，急性细胞死亡或慢性脑组织丢失，以及（3）持续（长达1年）仅在多次命中后出现认知缺陷19 （图2））。
弥漫相关光谱评估脑血流 弥漫相关光谱（DCS）是一种非侵入性光学技术，可测量血流量5，20，21。在DCS中，近红外光源放置在组织表面上。将检测器放置在与组织表面上的光源固定的距离处，以检测通过组织倍增散射的光（图3）。散射移动的红细胞会导致检测到的光强度随时间波动。一种称为相关扩散理论的简单分析模型用于将这些强度波动与血流指数相关联（CBFi， 图4）。虽然CBFi （cm2 / s）的单位不是传统的流量单位（mL / min / 100 g），但先前对小鼠的研究表明，CBFi 与通过标记为MRI21的动脉旋转测量的脑血流量密切相关。
作为参考，这里使用的DCS仪器是内部构建的，由852 nm长相干长度激光器，4个光子计数雪崩光电二极管阵列和硬件自相关器板（单tau，8通道，100 ns最小采样时间）21，22组成。数据是使用LabView编写的自制软件获取的。该器件的动物接口由 400 μm 多模源光纤（400-2200 nm 波长范围、纯硅芯、TECS 硬包层）和 780 nm 单模检测器光纤（780-970 nm 波长范围、纯硅芯、TECS 硬包层、730 ± 30 nm 秒模式截止）组成，间隔 6 mm 并嵌入黑色 3D 打印传感器 （4 mm x 8 mm， 图 3）。
神经炎症环境的量化 虽然神经炎症受多种细胞过程的调节，但两个关键的相关机制是细胞因子/趋化因子的细胞外信号传导和磷酸化蛋白的细胞内信号传导。为了研究损伤后大脑的神经炎症环境，从小鼠中提取大脑，进行微解剖，并使用Luminex量化细胞因子/趋化因子和磷酸化蛋白（图5， 图6， 图7）。Luminex多重免疫分析通过将酶联免疫吸附测定（ELISA）与荧光标记的磁珠偶联，能够同时对这些蛋白质的各种集合进行定量。对每种感兴趣的蛋白质使用不同的荧光标签，并且每个标签的磁珠都用针对该特定蛋白质的捕获抗体进行功能化。将数百个用于捕获每种蛋白质的磁珠混合在一起，置于96孔板中，并与样品一起孵育。样品孵育后，使用磁铁将磁珠捕获在孔中，同时将样品洗出。接下来，生物素化的检测抗体与感兴趣的分析物结合，形成类似于传统ELISA的抗体 - 抗原三明治，但每种蛋白质的ELISA发生在不同的荧光标记珠上。加入藻红素偶联链霉亲和素（SAPE）完成每个反应。然后，Luminex仪器读取磁珠并根据每个荧光标签/蛋白质分离信号。
数据集成 由于在Luminex测定中测量了大量的分析物（例如细胞因子），如果单独分析每个定量的蛋白质，则数据分析可能难以解释。为了简化分析并捕获分析物中观察到的趋势，我们使用一种称为偏最小二乘回归的多元分析方法（PLSR， 图8）23。PLSR的工作原理是识别对应于每个被测蛋白质（即细胞因子或磷酸化蛋白，称为“预测变量”）的权重轴，这些权重轴与反应变量（例如，脑血流量）一起最优地解释了被测蛋白质的协方差。权重被称为“载荷”，并被组装成一个称为潜在变量（LV）的向量。通过在两个LV中的每一个上投影（称为“评分”）测量的蛋白质数据，可以根据这些LV重新绘制数据。计算 PLSR 后，我们使用 varimax 旋转来识别新的 LV，该 LV 使样本投影到 LV 和预测变量24 之间的协方差最大化。这种方法允许我们将LV1定义为响应变量方差得到最好解释的轴。LV2最大化了响应变量和LV1残留数据之间的协方差，这可能与样品之间的生物或技术变异性有关。最后，我们进行一劳永逸的交叉验证（LOOCV），以确保PLSR模型不会严重依赖于任何一个样本23。
在该协议中，我们详细介绍了表征神经炎症和血流动力学组织对mTBI反应的方法。一般工作流程如图 1 所示。在该协议中，小鼠使用重量下降闭合性头部损伤模型进行一个或多个mTBI。脑血流量在损伤前和损伤后的多个时间点纵向测量。在询问神经炎症变化的兴趣点，动物被安乐死，大脑被提取出来。通过显微切割分离感兴趣的大脑区域，然后裂解以提取蛋白质。然后，裂解物用于细胞因子和磷酸化蛋白表达的Luminex多重免疫测定以及蛋白质印迹。最后，使用偏最小二乘回归分析集成该整体数据集。

<table>
	<tbody>
		<tr>
			<td>Adjustable pipettes</td>
			<td></td>
			<td></td>
			<td>any adjustable pipette</td>
		</tr>
		<tr>
			<td>Aluminum foil</td>
			<td>VWR</td>
			<td>89107-726</td>
			<td></td>
		</tr>
		<tr>
			<td>Bio-Plex cell lysis kit</td>
			<td>C Bio-Rad</td>
			<td>171304012</td>
			<td></td>
		</tr>
		<tr>
			<td>BRAND BRANDplates pureGrade Microplates, Nonsterile</td>
			<td>BrandTech</td>
			<td>781602 96</td>
			<td></td>
		</tr>
		<tr>
			<td>Complete mini protease inhibitor tablet</td>
			<td>Sigma-Aldrich</td>
			<td>11836153001</td>
			<td></td>
		</tr>
		<tr>
			<td>Depilatory cream</td>
			<td>Amazon</td>
			<td>Nair</td>
			<td></td>
		</tr>
		<tr>
			<td>DiH2O</td>
			<td>VWR</td>
			<td>VWRL0200-1000</td>
			<td></td>
		</tr>
		<tr>
			<td>Handheld magnetic separator block for 96 well flat bottom plates</td>
			<td>Millipore Sigma Catalogue</td>
			<td>40-285</td>
			<td></td>
		</tr>
		<tr>
			<td>Hardware Autocorrelator Board</td>
			<td>www.correlator.com</td>
			<td>Flex05-8ch</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane 250 mL</td>
			<td>MED-VET INTERNATIONAL</td>
			<td>RXISO-250</td>
			<td></td>
		</tr>
		<tr>
			<td>Kimwipe (11.2 x 21.3 cm)</td>
			<td>VWR</td>
			<td>21905-026</td>
			<td></td>
		</tr>
		<tr>
			<td>Laboratory vortex mixer</td>
			<td>VWR</td>
			<td>10153-838</td>
			<td></td>
		</tr>
		<tr>
			<td>LabView</td>
			<td>National Instruments</td>
			<td>LabVIEW</td>
			<td></td>
		</tr>
		<tr>
			<td>Luminex 200, HTS, FLEXMAP 3D, or MAGPIX with xPONENT software</td>
			<td>Luminex Corporation</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Luminex Drive Fluid</td>
			<td>Luminex</td>
			<td>MPXDF-4PK</td>
			<td></td>
		</tr>
		<tr>
			<td>Luminex sheath fluid</td>
			<td>EMD Millipore</td>
			<td>SHEATHFLUID</td>
			<td></td>
		</tr>
		<tr>
			<td>MILLIPLEX MAP Mouse Cytokine/Chemokine Magnetic Bead Panel - Premixed 32 Plex - Immunology Multiplex Assay</td>
			<td>Millipore Sigma</td>
			<td>MCYTMAG-70K-PX32</td>
			<td></td>
		</tr>
		<tr>
			<td>MILLIPLEX MAPK/SAPK Signaling 10-Plex Kit-Cell Signaling Multiplex Assay</td>
			<td>Millipore Sigma</td>
			<td>48-660MAG</td>
			<td></td>
		</tr>
		<tr>
			<td>Mini LabRoller rotator</td>
			<td>VWR</td>
			<td>10136-084</td>
			<td></td>
		</tr>
		<tr>
			<td>Phenylmethylsulfonyl fluoride</td>
			<td>Sigma-Aldrich</td>
			<td>P7626-1G</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate-buffered Saline (PBS)</td>
			<td>VWR</td>
			<td>97064-158</td>
			<td></td>
		</tr>
		<tr>
			<td>Plate Sealer</td>
			<td>VWR</td>
			<td>82050-992</td>
			<td></td>
		</tr>
		<tr>
			<td>Polypropylene microfuge tubes</td>
			<td>VWR</td>
			<td>20901-547</td>
			<td></td>
		</tr>
		<tr>
			<td>Mini LabRoller</td>
			<td>Millipore Sigma</td>
			<td>Z674591</td>
			<td></td>
		</tr>
		<tr>
			<td>Reagent Reservoirs</td>
			<td>VWR</td>
			<td>89094-668</td>
			<td></td>
		</tr>
		<tr>
			<td>R Programming Language</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>RStudio</td>
			<td>www.rstudio.com</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sonicator</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Titer plate shaker</td>
			<td>VWR</td>
			<td>12620-926</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween20</td>
			<td>Sigma-Aldrich</td>
			<td>P9416-50ML</td>
			<td></td>
		</tr>
		<tr>
			<td>1 m acrylic guide tube</td>
			<td>McMaster-Carr</td>
			<td>49035K85</td>
			<td></td>
		</tr>
		<tr>
			<td>4 photon counting avalanche photodiode</td>
			<td>Perkin-Elmer</td>
			<td>SPCM-AQ4C-IO</td>
			<td></td>
		</tr>
		<tr>
			<td>400 um multimode source fiber</td>
			<td>Thorlabs Inc.</td>
			<td>FT-400-EMT</td>
			<td></td>
		</tr>
		<tr>
			<td>54 g bolt</td>
			<td>Ace Hardware</td>
			<td></td>
			<td>0.95 cm basic body diameter, 2 cm head diameter, 10.2 cm length</td>
		</tr>
		<tr>
			<td>780 nm single mode detector fiber</td>
			<td>Thorlabs Inc.</td>
			<td>780HP</td>
			<td></td>
		</tr>
		<tr>
			<td>852 nm long-coherence length laser</td>
			<td>TOPTICA Photonics</td>
			<td>iBeam smart</td>
			<td></td>
		</tr>
	</tbody>
</table>

systems analysis of the neuroinflammatory and hemodynamic response to traumatic brain injury

轻度创伤性脑损伤（mTBI）是一个重大的公共卫生问题。反复暴露于 mTBI 可导致累积的、持久的功能缺陷。我们小组和其他人的大量研究表明，mTBI刺激细胞因子表达并激活小胶质细胞，减少脑血流量和新陈代谢，并损害脑血管反应性。此外，一些作品报告了这些神经炎症和血液动力学标志物中的紊乱与认知障碍之间的关联。在本文中，我们详细介绍了表征小鼠神经炎症和血流动力学组织对mTBI的反应的方法。具体而言，我们描述了如何执行mTBI的重量下降模型，如何使用称为弥漫相关光谱的非侵入性光学技术纵向测量脑血流量，以及如何对脑组织样品进行Luminex多重免疫测定以量化细胞因子和免疫调节磷酸化蛋白（例如，在MAPK和NFκB途径内），这些蛋白响应并调节小胶质细胞和其他神经免疫细胞的活性。最后，我们详细介绍了如何使用多变量系统分析方法集成这些数据，以了解所有这些变量之间的关系。了解这些生理和分子变量之间的关系最终将使我们能够确定导致mTBI的机制。

先前收集的数据取自先前的工作，其中一组8只C57BL / 6小鼠遭受三次闭合头部损伤（图2），间隔一次，每日11。在这项工作中，在最后一次损伤后4 h用弥漫相关光谱测量脑血流量（图3， 图4）。在损伤后CBF评估后，对动物进行安乐死，并通过免疫测定提取脑组织以定量细胞因子和磷酸化蛋白（图5</...

Watch this Scientific Journal Video about Systems Analysis of the Neuroinflammatory and Hemodynamic Response to Traumatic Brain Injury at JoVE.com

Systems Analysis of the Neuroinflammatory and Hemodynamic Response to Traumatic Brain Injury

该协议提出了表征神经炎症和血流动力学对轻度创伤性脑损伤的反应的方法，并将这些数据整合为使用偏最小二乘回归的多变量系统分析的一部分。

创伤性脑损伤的神经炎症和血流动力学反应的系统分析

Mild traumatic brain injuries (mTBIs) are a significant public health problem. Repeated exposure to mTBI can lead to cumulative, long-lasting functional ...

这些方法可用于表征神经炎症和血流动力学对脑损伤的反应，作为使用偏最小二乘回归的多变量系统分析的一部分。这些技术用于提供大脑对损伤反应的整体视图。为了诱导损伤，确认麻醉小鼠对脚趾捏合缺乏反应，并将小鼠置于薄膜中心的俯卧位。用双手握住所教组织，将鼠标的尾巴固定在拇指下，并将鼠标头定位在导管下方。当鼠标就位时，瞄准眼睛后部和耳朵前部之间的冲击力，将螺栓从导管顶部放到鼠标头部的背侧。撞击时，小鼠将突破组织，允许头部围绕颈部的快速加速。受伤后，让小鼠在37摄氏度的加热垫上以仰卧位恢复。稳定两分钟后，将DCS传感器轻轻地放在鼠标头骨的右半球上，使光学传感器的顶部边缘与眼睛的后部对齐，传感器的侧面沿着中线对齐。用杯子放在传感器上，以保护其免受室内光线的影响，并获取五秒钟的数据。然后，将传感器重新定位在左半球，并获取五秒钟的左半球数据。为了评估多重细胞因子和磷酸化蛋白的产生，每收获的动物脑组织约3毫克加入150微升混合裂解缓冲液。并使用1， 000微升移液器吸头对组织进行机械研磨15至20次。将均质化的样品在4摄氏度下在旋转器上放置30分钟，然后通过离心收集组织碎片。将上清液转移到新管中并离心样品以除去任何剩余的沉淀物。将上清液转移到新管中，并根据标准方案通过BCA测定测定蛋白质浓度。以新管中的线性范围分析确定的最佳蛋白质浓度制备每个样品的25微升。使用测定缓冲液将每个样品的总体积归一化至25微升。为了制备测定板，向96孔板的每个孔中加入200微升洗涤缓冲液，并在振荡器上以每分钟750转的速度摇动板10分钟。在孵育结束时，倾析洗涤缓冲液并将板敲击到纸巾上以除去任何残留物。接下来，向每个孔中加入25微升的测定缓冲液，然后向背景孔中加入25微升的缓冲液。将25微升稀释的样品加入适当的样品孔中，并涡旋多路复用磁珠的通孔一分钟，然后向每个孔中加入25微升完全重新悬浮的磁珠。添加完所有珠子后，用板封口机密封板，并用箔片盖住板，在两到八摄氏度下振荡过夜孵育。第二天早上，将板放在磁选机上，确保孔与磁铁对齐。两分钟后，用仍然附着在磁选机上的板倾析孔内容物，并向每个孔中加入200微升洗涤缓冲液。保持摇晃两分钟后，将板放在磁选机上两分钟。然后，用仍然附着在磁选机上的板倾析孔内容物，并用每孔200微升的洗涤缓冲液洗涤板，如刚才所示。第二次洗涤后，每孔加入25微升检测抗体，并用箔重新覆盖，在室温下以每分钟750转的速度孵育一小时。在孵育结束时，向每个孔中加入25微升链霉亲和素藻红素，并将板返回到振荡器中，在室温下再放置30分钟。在孵育结束时，将板放在磁选机上两分钟，然后倾析孔内容物。每次洗涤用200微升新鲜洗涤缓冲液洗涤板两次，并向每个孔中加入75微升适当的驱动液。然后，在室温下将磁珠重新悬浮在平板振荡器上五分钟，并根据制造商的说明在分析仪上读取板。在该分析中，在最后一次损伤后四小时用弥漫相关光谱测量脑血流量。然后可以确定每个半球的脑血流量指数和平均脑流量指数。在从所有样品收集数据之前，可以进行线性范围分析以确定适当的蛋白质上样质量。可以通过从样品数据中减去背景测量值，然后计算每种阳极解质的Z评分数据来制备细胞因子数据。部分最小二乘回归可以通过使用吞噬细胞小胶质细胞活化标记或其他感兴趣的变量作为响应变量，并将细胞因子测量作为预测变量来进行。可以执行Varimax旋转，以最大化潜在变量1上数据的协方差，并进行激活标记测量。潜伏变量1中的高负载权重对应于与活化标记物的高表达最相关的细胞因子表达。活化标志物和细胞因子之间的线性回归表明，在潜伏变量1中具有最大负载权重的细胞因子对于该分析也具有统计学意义。虽然我们把这个方案集中在创伤性脑损伤上，但这些方法被广泛推广到影响大脑的大量病理状况的研究上。我们使用这些技术来帮助我们确定可处理的靶点，以便在未来的实验中进行调节，以建立因果机制关系，并最终开发针对轻度创伤性脑损伤的新治疗策略。

Research

JoVE Journal

Neuroscience

Lateral Fluid Percussion: Model of Traumatic Brain Injury in Mice

侧向液压冲击：创伤性脑损伤小鼠模型

Traumatic brain injury (TBI) research has attained renewed momentum due to the increasing awareness of head injuries, which result in morbidity and ...

科研

神经科学

Investigations on Alterations of Hippocampal Circuit Function Following Mild Traumatic Brain Injury

调查轻度创伤性脑损伤海马电路功能的改变

Traumatic Brain Injury (TBI) afflicts more than 1.7 million people in the United States each year and even mild TBI can lead to persistent neurological ...

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

A Method to Inflict Closed Head Traumatic Brain Injury in Drosophila

方法对敌人造成闭合性颅脑创伤性脑损伤<em&gt;果蝇</em

Traumatic brain injury (TBI) affects millions of people each year, causing impairment of physical, cognitive, and behavioral functions and death. Studies ...

A Novel In Vitro Model of Blast Traumatic Brain Injury

A Novel In Vitro Model of Blast Traumatic Brain Injury

一种新的脑外伤体损伤模型

Traumatic brain injury is a leading cause of death and disability in military and civilian populations. Blast traumatic brain injury results from the ...

Effects of Blast-induced Neurotrauma on Pressurized Rodent Middle Cerebral Arteries

爆炸性神经损伤对压生啮鼠中脑动脉的影响

Though there have been studies on the histopathological and behavioral effects of blast exposure, fewer have been dedicated to blast&#39;s cerebral ...

Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice

研究小鼠创伤性脑损伤后凯库姆微生物群的改动

Increasing evidence shows that the microbiota-gut-brain axis plays an important role in the pathogenesis of brain diseases. Several studies also ...

Advanced Diffusion Imaging in The Hippocampus of Rats with Mild Traumatic Brain Injury

轻度创伤性脑损伤大鼠海马学的高级扩散成像

Mild traumatic brain injury (mTBI) is the most common type of acquired brain injury. Since patients with traumatic brain injury show a tremendous ...

Murine Model of Controlled Cortical Impact for the Induction of Traumatic Brain Injury

创伤性脑损伤诱导受控皮质冲击的母模型

The Centers for Disease Control and Injury Prevention estimate that almost 2 million people sustain a traumatic brain injury (TBI) every year in the ...

Intra-Arterial Delivery of Neural Stem Cells to the Rat and Mouse Brain: Application to Cerebral Ischemia

神经干细胞向大鼠和小鼠大脑的动脉内传递：对脑缺血的应用

Neural stem cell (NSC) therapy is an emerging innovative treatment for stroke, traumatic brain injury and neurodegenerative disorders. As compared to ...

A Metric Test for Assessing Spatial Working Memory in Adult Rats Following Traumatic Brain Injury

创伤性脑损伤后评估成年大鼠空间工作记忆的指标测试

Impairments to sensory, short-term, and long-term memory are common side effects after traumatic brain injury (TBI). Due to the ethical limitations of ...