这项工作是由来自德研究所Salud的卡洛斯三世赠款，通过PI10 / 00160和PI12 / 02839，部分被FEDER和穆图阿Madrileña支持。 AS-G。从穆图阿Madrileña一个博士后奖学金的获得者。三维仿真采用Bio数码Human软件（创建<a href="http://www.biodigital.com">www.biodigital.com</a> ）和ZygoteBody Professional软件（www.zygotebody.com）

在下面描述的协议中，步骤1,2和3属于两个研究和临床方案，这两者都是严格从仅通过临床标准选择的每个近中TLE候选切除手术，随后。步骤4和5专属于研究协议。这两个程序都是按照医院德拉公主伦理委员会的指导方针。 1.预植入程序<ol><li>解释了实验过程的参与者，指定哪个点对应的研究，哪些是适用于临床实践，陈述了研究过程中没有办法修改临床操作。要特别注意解释的电极植入手术的潜在风险。获取签署知情同意书的参与者。 </li><li>对于所有候选人切除手术，术前进行神经学和神经心理学EXAminations 18。 <ol><li> 20个国际系统与莫兹利的-根据10评估通过发作间期单光子发射计算机断层显像（SPECT）与99锝HMPAO，磁共振成像使用25头皮电极（MRI）1.5 T和视频脑电图（V-EEG）病人协议18。 </li><li>在手术前V形脑电图记录期间，逐步锥度从第二天的抗癫痫药物到第四天（每天的剂量大约三分之一）。 </li></ol></li></ol> 2.植入程序（手术） <ol><li>施用抗癫痫药预可操作，并在全身麻醉下进行手术（3毫克/公斤异丙酚丸剂，接着0.2 - 0.3毫克/千克芬太尼和0.5毫克/千克罗库溴铵）。 </li><li>将两个六敌人接触用1厘米的中心到中心的距离为双侧使用克氏针技术19周围的蓄水池。 <ol><li>将病人Øn个手术台仰卧位，与颈部在15度轻轻地延长。与碘溶液制备患者的面颊，开始在切口部位和向外盘旋，并悬垂立即切口部位周围的区域。 </li><li>根据哈特尔的地标20与20号脊椎针头穿刺皮肤:一个入口点约3cm侧向朝向一个点口服合缝立即劣于前-后平面同侧瞳孔和点大致的同侧2.5厘米前在横向平面外耳道。 </li><li>推进朝针透视下卵圆孔的区域。使用由透视图像提供横向的观点来确定针尖端的位置。当针通过卵圆孔，取出针芯，用电极取代它，并提前入环池（ 图1A </STRONG&gt;）。 </li></ol></li><li>评估通过在手术室21荧光透视成像正确注入;排除渗透到颅底，中椎间孔这一点很重要，如眶下裂（位于前部的卵圆孔）和颈静脉孔（位于后部的话）。这样的错位插管可能会导致严重的神经血管损伤22。 </li><li>一旦电极被正确地定位在环境蓄水池，它们固定到与窗帘皮肤。醒来后病人，并导致他或她的恢复室。 </li></ol> 3.录音FOE收购<ol><li>返回病人到V-脑电图室逗留约5.2±2.4天（平均值±SD）。 </li><li>根据国际10-20体系放置19电极。 <ol><li>用卷尺测量鼻根（鼻梁）和INION（枕骨粗隆）之间的距离，d标记与标记在中间点（锆石电极的位置）。测量并标记点的鼻根（所述FPZ电极的位置）以上的距离的10％。 <ol><li>重复同样的步骤为INION（盎司电极的位置），标志着距离从锆石20％，在鼻根和INION方向（分别为Fz的和PZ电极的位置）两者。 </li></ol></li><li>同时测量耳前点之间的距离，并标记的距离的左和右耳前点（T3和T4的电极，分别）在10％以上。然后，标记的距离的20％都T3和T4在直拉方向以上以获得C3和C4的位置。 </li><li>使用卷尺到FPZ和盎司在两个电极以上在前面和在01（左）和O2（右）在后面的距离在FP1（左）和FP2（右）的5％链接创建一个圆周上。 </li><li>在同一圆周上，在INION方向添加的距离的10％上升至OBT泉F7的位置，加10％，达到T3（它应位于耳前点之间的线的上方），并添加另外10％，得到的T5（O1电极）。标记各电极位置，并重复同样的程序为右（偶数）的电极。 </li><li>测量和标记中途F7和Fz的，并在F3方向从FP1向上距离的20％之间的交集（F3电极位置）。重复此过程中的头部的每个象限获得F4（前右位置），P3（左后位置）和P4（背左位置）。 </li><li>清洁和干燥皮肤。放置火棉胶的温和量与在各电极杯导电凝胶，并在坦然区域电极定位。干后用吹风机的火棉胶。 </li></ol></li><li>全部由电​​线电极（头皮和敌人）的连接电极中，这已经是连接到一个electroencephalographer。确保电极信号都不错，并验证头皮电极IMPE舞蹈是在使用electroencephalographer为10kΩ。 </li><li>获取数字头皮脑电图（EEG）数据和FOE-数据1,024赫兹使用视频同步electroencephalographer（ⅴ脑电图），和利用带通滤波器在0.5过滤数据 - 100赫兹和一个陷波滤波器（50赫兹）与electroencephalographer。 </li><li>逐步消除来自第二抗癫痫药物第四天（每天的剂量约三分之一），以增加癫痫发作的可能性。此步骤取决于每个病人的特定的药物处方。 </li><li>同时使用发作阵发性发作活动约通过识别电极/频道里癫痫元素出现23，包括慢复合波，多峰，快速棘波，尖波，尖和慢复合波的运行定位ictogenic区慢尖波，尖峰和棘慢波。记录发作起始和结束的时间，以及一个Y以外的临床症状或出现相关的研究。有在患者的头部的电极的位置和在其允许解剖识别所述致癫痫活性的显示位置脑电图软件的头部模型之间的一对一的映射。 </li><li>当研究完成后，轻轻地向外拉出来，而病人的口腔保持半开除去在V-EEG单位的敌人。 FOE删除后没有系统地进行拍摄，当出现神经系统症状时除外。在这样的情况下，执行紧急计算机断层扫描（CT）扫描。 </li></ol> 4. FOE信号预处理<ol><li>导出在200Hz上存储electroencephalographer ASCII格式以适合于大约30分钟癫痫发作活动的（已经由专家神经生理学家标识）（ 图1C）的数值分析历元中的数据。避免含工件历元，如饱和电活动，肌肉活动，以及electrode位移。 </li><li>打开使用任何UNIX流编辑器导出的文件，并从导出的数据文件中删除所有的非数字字符，只留下时间戳和通道电压。除进一步的数值分析，修改后的文件。 注:从现在起，执行使用R封装从R库或自制码（ 表1）所有计算。 </li><li>基于R软件，安装所需的R程序包，以及修改后的数据文件加载到第r环境。订购的所有通道，指定每一个到包含的所有数据，消除空渠道，他们引用的平均中线的参考法（FZ +锆+ Pz的）/ 3阵列的特定列。 <ol><li>使用快速傅立叶变换算法（R函数:FFT）并绘制得到的变量，以检查有效去除线频率（大约50Hz）的。使用频域来过滤其他杂散FRequencies可能污染信号。 </li></ol></li><li>加载的数据转换为一个多元时间序列对象28列（MTS） - 16头皮和12的敌人-使用R功能的TS。划分MTS对象成每次5秒（在200赫兹1000个数据点）的非重叠时间窗口以减少文件大小和优化计算时间。 </li></ol> 5.后处理计算（复杂网络分析） 注:计算在每个时间窗口下面所述的措施，开始痉挛发作（60窗口）前5分钟，在痉挛发作后5分钟（60窗口）结束，用可视化的时空演化的目的。 <ol><li>计算单变量的措施，光谱功率，兴奋性和谱熵为每个列/通道不考虑不同时间序列之间的关系。 <ol><li>计算兴奋（S）为各V使用根据迅达24提出的公式自制代码电压活动时间系列（见补充文件）。 S&gt; 2.5被认为是癫痫，经验确定的阈值17,25,26。 </li><li> （ - 7赫兹4），阿尔法（7 - 14赫兹），β（14 - 30赫兹为每个活动的时间序列，使用自制的代码为德尔塔（&gt; 0.5赫兹和&lt;4赫兹），西塔计算功率谱密度）和伽玛（&gt; 30）。 </li><li>计算香农熵使用每个时间序列代替的相应概率的时间序列的功率谱密度的自制代码。平均超过一组电极每一个信道得到的各个频谱熵（SE）的值。香农熵在补充文件中说明。 注:在SE的减少应该被解释为在光谱的频率的数量的减少，因为SE是光谱的熵。 </li></ol></li><li> 网络措施 不E:本节评估不同的时间序列的电极之间的相互作用。 <ol><li>利用在零延迟（:CCF R函数）计算出的线性交叉相关系数的绝对值计算在每个时间窗口中的每对电压的时间序列之间的功能连接。 注:要消除同步的非代表值，建立了基于以往的研究17,25,26的阈值。在这种特殊情况下使用的0.5的阈值。 </li><li>安装的igraph - [R包27。创建一个从邻接矩阵的IGRAPH对象（R功能:graph.adjacency）。使用在先前步骤中获得的相关矩阵，指定的图形被加权和无向。 </li><li>在每个时间窗口中计算整个网络（头皮+ FOE）平均路径长度（APL）（R函数average.path.length），并为每个所述四个子网络:左头皮，右头皮，左FOE和正确的敌人。在电子Xactly公司以同样的方式，计算链接密度（DOL）（R功能:graph.density），模块化（MOD）（R功能:模块化），平均聚集系数（ACC）（R功能:传递）。 </li><li>重复使用相位同步（自制R代码里面 ），为功能连接，而不是互相关函数的估计的先前步骤5.2.1至5.2.3。 </li></ol></li><li>来表示变量的大小变化的影响，计算出标准化平均差（SMD）（R函数从包MBESS:SMD），所述preictal和发作阶段之间以及preictal和发作后阶段之间。 <ol><li>服用preictal为基线，选择30秒钟（6个值），五分钟前发作起始标志，作为preictal值。可以在癫痫发作过程中以量化的变化，对于preictal阶段，通过使用SMD选择的30个类似的时间窗口。 </li><lI&gt;以类似的方式，发作结束5分钟后，以量化在发作后阶段（在相对于preictal阶段）的变化选择的30秒的时间窗口。 </li></ol></li></ol>

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G., Pastor, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+mesial+synchronization+in+temporal+lobe+epilepsy.">Impaired mesial synchronization in temporal lobe epilepsy.</a> Clin Neurophysiol. 122 (6), 1106-1116 (2011).</li><li>Bettus, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Decreased+basal+fMRI+functional+connectivity+in+epileptogenic+networks+and+contralateral+compensatory+mechanisms.">Decreased basal fMRI functional connectivity in epileptogenic networks and contralateral compensatory mechanisms.</a> Hum Brain Mapp. 30 (5), 1580-1591 (2009).</li><li>Pereira, F. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asymmetrical+hippocampal+connectivity+in+mesial+temporal+lobe+epilepsy:+evidence+from+resting+state+fMRI.">Asymmetrical hippocampal connectivity in mesial temporal lobe epilepsy: evidence from resting state fMRI.</a> BMC Neurosci. 11, 66 (2010).</li><li>Vega-Zelaya, L., Pastor, J., Tormo, I., de Sola, R. G., Ortega, G. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessing+the+equivalence+between+etomidate+and+seizure+network+dynamics+in+temporal+lobe+epilepsy.">Assessing the equivalence between etomidate and seizure network dynamics in temporal lobe epilepsy.</a> Clin Neurophysiol. 127 (1), 169-178 (2011).</li><li>Pastor, J., Sola, R. G., Ortega, G. 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The authors have no conflicts of interest to disclose.

传统上，癫痫是一种面向区域的方法，其中分离出特定领域，基本上是发作起始区的重要性，因为癫痫发作的病因独特涉猎。最近，一种强调皮层区之间相互作用的重要性一个真正的网络方法已被青睐在经典的面向区域的透视13-17,28。然而，癫痫作为网络疾病的证据目前身体仍然高度分散，而且需要更多的研究。目前的工作旨在通过重新分析传统方法作为FOE提供的数据，复杂的网络方式下。这里介绍的协议描述一个循序渐进的方法步骤，在TLE患者进行半创记录的复杂网络和频谱分析。 上面描述的技术的应用已表明了网络的方法的有用性是与更传统的禄相比alized或面向区域的观点。在最近的作品17,29已表明，使用同样的步骤，这里所描述的，在近中连接在耐火TLE患者的不平衡是显而易见的。近中连接在同侧无论是在发作间期29和发作17,29阶段减少。这个结果不能仅仅通过在那里的致癫痫活性发生的地区寻找可以预见。这在某种程度上令人惊讶的结果也利用功能磁共振成像上信号30,31网络理论描述。此外，FOE +网络理论的组合技术的应用已经显示出近中活动的等价期间发作和下致癫痫活性的启动子的影响，因为它是依托咪酯32的药理学施用。 此处所描述的技术能够在短间期录音检测近中网络不平衡至多一个O持久- [R2小时29。以这种方式，在分析中的时间和病人住院时间大幅减少，可以实现的。另外，从治疗的角度看，在TLE患者现有的不平衡可以通过使用长期植入（由神经外科医生）设备，多，因为它是在深部脑刺激做的方式"解决"。 要使用在本协议中提供的信息，取得最佳的效果，有些问题应提前考虑。首先，电极植入应由有经验的神经外科医生来进行，因为他们的不正确的位置可能会产生严重的神经系统的后果和误导性的记录。此外，为进一步分析相应时期的选择完全依赖于原始脑电图的神经生理学家的解释;因此，在临床脑电图分析方面的经验是强制性的。从脑电图导出文件的数据格式取决于汉邦丘拉尔品牌;因此，需要良好的编程技能相适应的脚本以不同的数据格式。最后，为了确保数据的可靠性，质量控制应该被应用到的结果。过度估计和误报可能具有大量的相关性的工作时出现。在这种情况下，统计方法，以提高灵敏度，应使用。在这方面，必须建立在相关性的阈值丢弃并不代表真正的底层同步的值是重要的。因此，在该协议中，节点i和j之间的边缘将仅视为存在如果这些节点之间的相关性的绝对值大于0.5，以前使用17,26的标准。在0.2至0.8的范围内的其他的阈值应该被用来验证相似的结果，并确保从一个阈值到以下阈值的平滑过渡。除了阈值，其他methodologie之间可以用于获得可靠的结果，如Bonferroni校正或替代数据的测试。此外，脑电图数据时，必须记住，大脑网络与非线性动力学复杂的系统是非常重要的;因此，除了线性相关，应使用其它非线性同步措施，以确保结果，诸如交互信息或相位同步33的质量。 直接从头皮上的电极连接的计算，因为它是在这项工作完成了一部分可能会带来一定的风险。主要的问题，休息，由于体积传导，总是存在用头皮记录污染的效果。克服这个问题的一种方式是通过在源的空间，由许多研究中采用的吸引人的选择工作。另一种方法要求使用同步措施的影响幅度污染降至最低。通过使用相位同步（也称为相大号ocking值）我们最小化体积传导的影响，因为它是在几个作品34证实。 如在其他侵入性神经生理技术，从FOE记录不能从对照受试者得到，这一事实严重地限制使用某些研究协议。从FOE录音数据提供有关内侧颞叶活动17,29,35有价值的信息，尤其是不对称的致痫侧颞叶癫痫病人33时。与侵入技术相比，FOE技术是非外伤性的脑并涉及相对简单的操作，其记录的高品质在一段11长时间。相较于MRI，FOE录音提供electrocortical活动更好的时间分辨率。此外，存在许多可能性，探索比在这项工作中使用的其他措施。这些事实也增加了一些分析生物医学记录的可能性同时。 FOE录音用复杂的网络和频谱分析相结合的这些优点使得该技术对癫痫研究与临床实践中的应用潜力的强大工具。

癫痫是一种致残性疾病，影响1 - 世界人口的2％。在大多数情况下，癫痫发作 - 癫痫的标志 - 可以完全控制或者与抗癫痫药物废除。然而，癫痫专利约30％的难治性药物治疗。在最常见的类型癫痫，颞叶癫痫（TLE）1，幸运的是手术是改善病人的状况的有效替代。从荟萃分析的结果表明，几乎三分之二耐药TLE患者是无癫痫发作在切除手术2,3后的第一个两到三年，虽然这一比例在几个因素而变化，最值得注意的是，海马的类型硬化2。对于一个成功的结果的一个关键步骤是所谓的癫痫病灶的准确定位，皮层区负责发作的产生，其通常位于梅西亚颞叶l的面积。然而，即使在癫痫病灶已在手术过程中被正确识别和切除的情况下，患者的一个显著少数或者与手术后癫痫保持或必须在严格的抗癫痫药治疗控制发作被放置。因此，一个新的观点却出现了注意力不再仅仅关注偏远地区，而不是皮质的相互作用构成了目前的根本问题。这种"网络"方法在连接组概念4，重点关注在不同区域之间的神经连接，而不是突出条块结构的作用接地。这一新的模式是在图论，专门用于图形拓扑和统计特性研究的一个数学框架中发现，适当的工具来表达它的基本结论。根据这种观点，大脑被认为是通过链路互连的一组节点<suP&gt; 5-9，使得节点通过由电极和它们之间的链路所覆盖的皮层区域被同步的程度定表示。因此，该网络的方法已在侵入电极记录的分析中使用，并提供了新的信息，以促进发作产生和传播的作用机制的理解。 在世界上大多数的癫痫中心常规采用的众多微创神经生理学技术，卵圆孔电极（FOE）尤为显着。 FOE是半侵入性技术，因为没有必要执行开颅，这减少了手术相关的并发症10。此外，FOE在环池11的位置，使它们从参与发作产生和传播几个皮质结构，如内嗅皮层记录近中活性特别方便。因此，它因为使用它的外观是耐药TLE患者的术前评估普遍。传统上，这种技术用于定位在发作癫痫尖峰和尖锐波的形式刺激活性，更重要的是，要准确地识别近中发作起始的区域。 从委员会提议的分类和术语从对癫痫国际联盟（ILAE）的新定义表明，癫痫发作起源于特定网络12内的一些点。此外，几个研究已经证明，癫痫发作是由异常的网络活动，而不是由一个孤立的病变区域13-16引起的。显然，这个新的角度使用新的数值的方法，如复杂的网络方法需要的先前获得的信息再分析。虽然这些分析的实际使用是在临床实践中仍然初期，几项研究已经证实其值13-17。 下面描述的协议是对耐药TLE癫痫病人常规进行与网络分析的新技术在临床实践的结合。该方法允许对近中的网络参数的时间演化的评估。 FOE的双边插入环池在颞叶的几个领域近中同时记录electrocortical活动。施加到记录时间序列的网络的做法既interictally和癫痫发作期间跟踪近中网络的时间演化。以这种方式，所提出的协议提供了可视化和量化考虑几个近中区域之间的关系的措施的独特方式。

<table border="0" cellpadding="0" cellspacing="0" width="655">
	<tbody>
		<tr height="63">
			<td height="63" style="height:63px;width:247px;">Foramen Ovale Electrodes</td>
			<td style="width:81px;">AD-Tech, Racine, 
			USA</td>
			<td style="width:136px;">FO06K-SP10X-000</td>
			<td style="width:191px;">Six-contact platinum&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Electroencephalograph</td>
			<td style="width:81px;">XLTEK, Canada</td>
			<td style="width:136px;">XLT-EEG32T</td>
			<td>Natus XLTEK</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:247px;">MRI machine</td>
			<td style="width:81px;">General Electric</td>
			<td style="width:136px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:247px;">SPEC machine</td>
			<td style="width:81px;">General Electric</td>
			<td style="width:136px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

network analysis of foramen ovale electrode recordings in drug-resistant temporal lobe epilepsy patients

癫痫患者的约30％是难治抗癫痫药。在这些情况下，外科手术是消除/控制癫痫发作的唯一选择。但是，患者的一个显著少数继续显示手术后癫痫，即使在癫痫发作的可疑源已被正确本地化和切除这些情况。这里介绍的协议相结合的颞叶癫痫（TLE）的患者的术前评价与网络分析的新技术中常规使用的临床过程。该方法允许对近中的网络参数的时间演化的评估。卵圆孔电极（FOE）的双边插入环池在颞叶数近中区域同时记录electrocortical活性。此外，施加到记录时间系列网络方法跟踪近中网络的时间演化既interictally并在癫痫发作。以这种方式，所提出的协议提供了可视化和量化考虑几个近中的区域，而不是一个单一的区域之间的关系的措施的独特方式。

的FOE的最终位置是在环池，如在轴向和矢状MRI（ 图1A上图 ）看出。仇敌记录电活动从颞叶（ 图1A下图 ）的几个近中结构的接触。手术（ 图1B左图 ）后，患者被调度到视频脑电图室，头皮电极与10根据置于- 20系统（ 图1B右侧 ）。期间留在视频脑电图室，病人被连续监视，保存用于进一步分析头皮和FOE-录音，以及视频和重要常数。一个典型的原始头皮和FOE信号（ 图1C）显示了扣押左侧FOE的外观和其散布于头皮和右FOE联系人。 通过使用致痫活动的表示兴奋性的（S）从preictal到发作和发作后时期的过渡过程中对应于从图1C，原始脑电图记录（ 图2）。发作起始标记用实垂直线和时间（x轴）都提到这一点。 S（兴奋）&gt;的值2.5表示刺激性或致癫痫活性17,25,26。兴奋性较高（偏红色调）与左侧FOE触点（LFOE）较高的强度首先出现。作为由专家神经生理学家获悉这一结果是一致的左颞叶癫痫。 的几个网络措施时间动态以及从preictal到发作和发作后的阶段，对应于图1C和图2中显示的相同的发作在过渡期间的频谱熵（ 图3）。发作起始标有一个坚实的VERTI校准线和时间（x轴）都提到这一点。在这种情况下，该网络在全组电极，包括头皮和FOE-建造。 DOL和ACC值癫痫发作时均较高，随着APL和MOD的下降，表明整体连通的增加。在此期间还进行了观察和持续的兴奋性（垂直虚线）消失后SE水平较低。 的网络措施ACC，DOLS和APL和SE每个FOE（右和左）（ 图4）从preictal到发作和发作后阶段的过渡期间，分析。发作起始标记用实垂直线和时间（x轴）都提到这一点。这种措施的演进对应于相同的发作图1，图2和3。同侧（左）近中ACC，DOLS和APL提出较早，较contrala高变化或双侧的值，这可能是由在发作起始区的位置在左颞叶进行说明。在这种情况下，国防部不能计算，因为没有细分可用。 功能连接相同扣押图1，2，3时（ 图5），4代表视频呈现只是痉挛发作（时间0）后的一个关键的变化。在这一点，所有电极之间的连接显着增加，如通过增加的链路的数目和厚度（强度）的可见的那个边缘。这种增加左FOE之间开始于时间0.1和0.2，并在到达整个网络之前扩散到对侧。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> FFT </td><td> 4.3 </td><td> （统计包）计算快速傅立叶TR的信号的ansform。 </td></tr><tr><td> TS </td><td> 4.4 </td><td> （统计包）创建一个多元时间序列对象（MTS）。应提供的采样频率。 </td></tr><tr><td>刺激性</td><td> 5.1.1 </td><td>基于差异 R函数（自制）的功能。计算的信号的斜率的绝对值，然后将它规范化为标准偏差短基线时期。应提供的阈值。 </td></tr><tr><td>功率谱密度和谱熵</td><td> 5.1.2 </td><td>基于频谱和熵 R里面的函数（自制）的功能。计算归一化的功率谱和归一化的功率谱的香农熵</td></tr><tr><td> CCF </td><td> 5.2.1 </td><td> （基本软件包）通过使用Pearson相关零滞后，产生一个相关矩阵计算山对象的线性互相关。绝对值应该演算ated。 </td></tr><tr><td> graph.adjacency </td><td> 5.2.2 </td><td> （IGRAPH包）创建的igraph图，通过以下的igraph功能使用的基本对象</td></tr><tr><td> average.path.length </td><td> 5.2.3 </td><td> （的igraph包）确定的曲线图的平均路径长度，通过所有网络节点的计算的沿最短路径的步骤的平均数目。 </td></tr><tr><td> graph.density </td><td> 5.2.3 </td><td> （的igraph封装）通过计算链接的实际数量和网络的所有可能的链路之间的比率计算的曲线图的链接的密度。 </td></tr><tr><td> 模块化 </td><td> 5.2.3 </td><td> （的igraph包）确定图形的模块化，其中节点组更它们之间连接比与网络的其他节点计算</td></tr><tr><td> 传递 </td><td> 5。2.3 </td><td> （的igraph包）确定的曲线图的平均聚类系数，通过计算也彼此的邻居的相邻节点的比例</td></tr><tr><td>相位同步</td><td> 5.2.4 </td><td> （自制）函数基于在计算平均相位相干性来0和1之间获得的值的fft R函数</td></tr><tr><td> SMD </td><td> 5.3 </td><td> （MBESS包）确定标准平均差-size effects-通过计算相对集中的差异群体之间平均的差异</td></tr></tbody></table> 表1:R功能，用于数据处理。 <img alt="图1" src="/files/ftp_upload/54746/54746fig1.jpg" /> 图1:卵圆孔电极。 （A）的最终位置FOE进入环池。上图显示的轴向（左）和矢状（右图）MRI图像显示FOE接触位置（白色箭头）。一个人体标本（尸体）与插入的FOE（下图，标有白色箭头触点）。 （B）FOE和头皮电极设置。患者只是FOE插入手术（左图）之后和视频脑电图住宿（右图）中的头。 （C）FOE和头皮录音。从左侧颞叶癫痫患者复杂部分性发作（5分钟后痉挛发作之前）。 RFOE1-RFOE6手段右FOE＃1至＃6和LFOE1-LFOE6手段左FOE＃1至＃6。发作起始由垂直红线和白色箭头标注的头。 <a href="http://ecsource.jove.com/files/ftp_upload/54746/54746fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/54746/54746fig2.jpg" /> 图2:从兴奋量化为左颞叶癫痫病人的一个复杂部分发作的表示。色阶量化每个电极的兴奋性电平（S）。右卵圆孔电极（RFOE）和左卵圆孔电极（LFOE）分别代表的右触点和左卵圆孔电极（y轴）。 x轴标志着时间（分钟）相对于痉挛发作（粗垂直线）由专家确定的神经生理学家。 <a href="http://ecsource.jove.com/files/ftp_upload/54746/54746fig2large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图3" src="/files/ftp_upload/54746/54746fig3.jpg" /> 图3: 整个网络（头皮+ FOE）从图2的平均聚集系数（ACC），平均路径长度同一患者同一和扣押措施 （APL），链接（DOLS），模块化（MOD）和整个网络（头皮+ FOE）谱熵（SE）密度表示。垂直虚线表示的兴奋性（S）。 x轴标志着时间相对痉挛发作（垂直粗实线）。在连续十窗户移动平均线是由一个厚厚的黑色实线表示。 <a href="http://ecsource.jove.com/files/ftp_upload/54746/54746fig3large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图4" src="/files/ftp_upload/54746/54746fig4.jpg" /> 图4:从图2和3的平均聚集系数（ACC），平均路径长度（APL） 同一患者的近中的措施 ，为左和右孔两条链路（DOLS）和频谱熵（SE）密度未闭的电极（敌人）。垂直虚线标记的兴奋性。 x轴相对于与s的时间标记eizure发作（垂直粗实线）。在连续十窗户移动平均线是由一个厚厚的黑色实线表示。 <a href="http://ecsource.jove.com/files/ftp_upload/54746/54746fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" src="/files/ftp_upload/54746/54746fig5.jpg" /> 图5: 动态连接模式的复杂部分发作期间。链接强度由边缘的厚度来表示。时报讯（数字越小），是相对于痉挛发作（时间0）。每一帧是5秒长。左，右卵圆孔电极（L1-L6和R1-R6）是由珊瑚和蓝色圆圈，分别代表。左，右头皮电极由橙色和青色圆圈，分别代表。 <a href="http://ecsource.jove.com/files/ftp_upload/54746/Figure_5.mp4" target="_blank">请点击此处到D</a>ownload这部电影。

Watch this Scientific Journal Video about Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients at JoVE.com

Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

在耐药性颞叶癫痫患者卵圆孔电极记录的网络分析

Approximately 30% of epilepsy patients are refractory to antiepileptic drugs. In these cases, surgery is the only alternative to eliminate/control ...

network-analysis-foramen-ovale-electrode-recordings-drug-resistant

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Pharmacology

Medicine

Unit 1

Pharmacotherapy of the Epilepsies

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12.1K 次观看.  Instituto de Investigación Sanitaria Hospital de la Princesa. This protocol describes a procedure to track the evolution of mesial network measures in temporal lobe epilepsy (TLE) patients. It is based on the combination of intracranial recordings with a novel numerical technique for data analysis. Specifically, we present a protocol for network analyses of foramen ovale recordings.

视频: Network Analysis of Foramen Ovale Electrode Recordings in Drug-resistant Temporal Lobe Epilepsy Patients

Generation of Subcutaneous and Intrahepatic Human Hepatocellular Carcinoma Xenografts in Immunodeficient Mice

In vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate the human disease provide a valuable platform for research into disease pathophysiology and for the preclinical evaluation of novel therapies. We present a variety of methods to generate subcutaneous or orthotopic human HCC xenografts in immunodeficient mice that could be utilized in a variety of research applications. With a focus on the use of primary tumor tissue from patients undergoing surgical resection as a starting point, we describe the preparation of cell suspensions or tumor fragments for xenografting. We describe specific techniques to xenograft these tissues i) subcutaneously; or ii) intrahepatically, either by direct implantation of tumor cells or fragments into the liver, or indirectly by injection of cells into the mouse spleen. We also describe the use of partial resection of the native mouse liver at the time of xenografting as a strategy to induce a state of active liver regeneration in the recipient mouse that may facilitate the intrahepatic engraftment of primary human tumor cells. The expected results of these techniques are illustrated. The protocols described have been validated using primary human HCC samples and xenografts, which typically perform less robustly than the well-established human HCC cell lines that are widely used and frequently cited in the literature. In comparison with cell lines, we discuss factors which may contribute to the relatively low chance of primary HCC engraftment in xenotransplantation models and comment on technical issues that may influence the kinetics of xenograft growth. We also suggest methods that should be applied to ensure that xenografts obtained accurately resemble parent HCC tissues.

Human tumor xenografts in immunodeficient mice are valuable tools to study cancer biology. Specific protocols to generate subcutaneous and intrahepatic xenografts from human hepatocellular carcinoma cells or tumor fragments are described. Liver regeneration induced by partial hepatectomy in recipient mice is presented as a strategy to facilitate intrahepatic engraftment.

皮下及肝内人肝癌裸鼠移植瘤的免疫缺陷小鼠的产生

In  vivo experimental models of hepatocellular carcinoma (HCC) that recapitulate  the human disease provide a valuable platform for research into disease  ...

科研

医学

Network Analysis of the Default Mode Network Using Functional Connectivity MRI in Temporal Lobe Epilepsy

Functional connectivity MRI (fcMRI) is an fMRI method that examines the connectivity of different brain areas based on the correlation of BOLD signal fluctuations over time. Temporal Lobe Epilepsy (TLE) is the most common type of adult epilepsy and involves multiple brain networks. The default mode network (DMN) is involved in conscious, resting state cognition and is thought to be affected in TLE where seizures cause impairment of consciousness. The DMN in epilepsy was examined using seed based fcMRI. The anterior and posterior hubs of the DMN were used as seeds in this analysis. The results show a disconnection between the anterior and posterior hubs of the DMN in TLE during the basal state. In addition, increased DMN connectivity to other brain regions in left TLE along with decreased connectivity in right TLE is revealed. The analysis demonstrates how seed-based fcMRI can be used to probe cerebral networks in brain disorders such as TLE.

The Default Mode Network (DMN) in Temporal Lobe Epilepsy (TLE) is analyzed in the resting state of the brain using seed-based functional connectivity MRI (fcMRI).

使用功能性磁共振成像连接在颞叶癫痫默认模式网络的网络分析

Functional connectivity MRI (fcMRI) is an fMRI method that examines the connectivity of different brain areas based on the correlation of BOLD signal ...

Multi-electrode Array Recordings of Human Epileptic Postoperative Cortical Tissue

Epilepsy, affecting about 1% of the population, comprises a group of neurological disorders characterized by the periodic occurrence of seizures, which disrupt normal brain function. Despite treatment with currently available antiepileptic drugs targeting neuronal functions, one third of patients with epilepsy are pharmacoresistant. In this condition, surgical resection of the brain area generating seizures remains the only alternative treatment. Studying human epileptic tissues has contributed to understand new epileptogenic mechanisms during the last 10 years. Indeed, these tissues generate spontaneous interictal epileptic discharges as well as pharmacologically-induced ictal events which can be recorded with classical electrophysiology techniques. Remarkably, multi-electrode arrays (MEAs), which are microfabricated devices embedding an array of spatially arranged microelectrodes, provide the unique opportunity to simultaneously stimulate and record field potentials, as well as action potentials of multiple neurons from different areas of the tissue. Thus MEAs recordings offer an excellent approach to study the spatio-temporal patterns of spontaneous interictal and evoked seizure-like events and the mechanisms underlying seizure onset and propagation. Here we describe how to prepare human cortical slices from surgically resected tissue and to record with MEAs interictal and ictal-like events ex vivo.

We here describe how to perform multi-electrode array recordings of human epileptic cortical tissue. Epileptic tissue resection, slice preparation and multi-electrode array recordings of interictal and ictal events are demonstrated in detail.

人类癫痫术后皮层组织的多电极阵列录音

Epilepsy, affecting about 1% of the population, comprises a group of neurological disorders characterized by the periodic occurrence of seizures, which ...

Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with focal epilepsy. This methodology relies on the assumption that seizures originate from the same regions of the brain from which interictal epileptiform discharges (IEDs) emerge. Preclinical models are very useful to find correlates between IED locations and the actual regions underlying seizure initiation in focal epilepsy. Rats have been commonly used in preclinical studies of epilepsy1; hence, there exist a large variety of models for focal epilepsy in this particular species. However, it is challenging to record multichannel EEG and to perform brain source imaging in such a small animal. To overcome this issue, we combine a patented-technology to obtain 32-channel EEG recordings from rodents2 and an MRI probabilistic atlas for brain anatomical structures in Wistar rats to perform brain source imaging. In this video, we introduce the procedures to acquire multichannel EEG from Wistar rats with focal cortical dysplasia, and describe the steps both to define the volume conductor model from the MRI atlas and to uniquely determine the IEDs. Finally, we validate the whole methodology by obtaining brain source images of IEDs and compare them with those obtained at different time frames during the seizure onset.

This video introduces the preparation, recording, and source analysis procedures of high-resolution EEG on sedated rats with a particular preclinical model of focal epilepsy under noninvasive conditions.

采用高分辨率脑电图记录脑源成像局灶性癫痫的临床前模型大鼠

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with ...

A Multimodal Imaging- and Stimulation-based Method of Evaluating Connectivity-related Brain Excitability in Patients with Epilepsy

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations over time in the blood-oxygenation level dependent signal. rs-fcMRI has been applied extensively to identify abnormalities in brain connectivity in different neurologic and psychiatric diseases. However, the relationship among rs-fcMRI connectivity abnormalities, brain electrophysiology and disease state is unknown, in part because the causal significance of alterations in functional connectivity in disease pathophysiology has not been established. Transcranial Magnetic Stimulation (TMS) is a technique that uses electromagnetic induction to noninvasively produce focal changes in cortical activity. When combined with electroencephalography (EEG), TMS can be used to assess the brain's response to external perturbations. Here we provide a protocol for combining rs-fcMRI, TMS and EEG to assess the physiologic significance of alterations in functional connectivity in patients with neuropsychiatric disease. We provide representative results from a previously published study in which rs-fcMRI was used to identify regions with abnormal connectivity in patients with epilepsy due to a malformation of cortical development, periventricular nodular heterotopia (PNH). Stimulation in patients with epilepsy resulted in abnormal TMS-evoked EEG activity relative to stimulation of the same sites in matched healthy control patients, with an abnormal increase in the late component of the TMS-evoked potential, consistent with cortical hyperexcitability. This abnormality was specific to regions with abnormal resting-state functional connectivity. Electrical source analysis in a subject with previously recorded seizures demonstrated that the origin of the abnormal TMS-evoked activity co-localized with the seizure-onset zone, suggesting the presence of an epileptogenic circuit. These results demonstrate how rs-fcMRI, TMS and EEG can be utilized together to identify and understand the physiological significance of abnormal brain connectivity in human diseases.

Resting-state functional-connectivity MRI has identified abnormalities in patients with a wide range of neuropsychiatric disorders, including epilepsy due to malformations of cortical development. Transcranial Magnetic Stimulation in combination with EEG can demonstrate that patients with epilepsy have cortical hyperexcitability in regions with abnormal connectivity.

评价中的连接有关的大脑兴奋患者癫痫多式联运Imaging-和激励为基础的方法

Resting-state functional connectivity MRI (rs-fcMRI) is a technique that identifies connectivity between different brain regions based on correlations ...

A Detailed Protocol for Physiological Parameters Acquisition and Analysis in Neurosurgical Critical Patients

Intracranial pressure (ICP) monitoring is now widely used in neurosurgical critical patients. Besides mean ICP value, the ICP derived parameters such as ICP waveform, amplitude of pulse (AMP), the correlation of ICP amplitude and ICP mean (RAP), pressure reactivity index (PRx), ICP and arterial blood pressure (ABP) wave amplitude correlation (IAAC), and so on, can reflect intracranial status, predict prognosis, and can also be used as guidance of proper treatment. However, most of the clinicians focus only on the mean ICP value while ignoring these parameters because of the limitations of the current devices. We have recently developed a multimodality monitoring system to address these drawbacks. This portable, user-friendly system will use a data collecting and storing device to continuously acquire patients&#39; physiological parameters first, i.e., ABP, ICP, and oxygen saturation, and then analyze these physiological parameters. We hope that the multimodality monitoring system will be accepted as a key measure to monitor physiological parameters, to analyze the current clinical status, and to predict the prognosis of the neurosurgical critical patients.

Recently, we have developed a portable multimodality monitoring system for the monitoring of various physiological parameters in neurosurgical critical patients. Detailed protocols on how to use this multimodality monitoring system are presented here.

神经外科危重患者生理参数采集与分析的详细方案

Intracranial pressure (ICP) monitoring is now widely used in neurosurgical critical patients. Besides mean ICP value, the ICP derived parameters such as ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

小鼠颞叶癫痫的 Radiotelemetry 模型及脑电图监测

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

Assessment of Static Graviceptive Perception in the Roll-Plane using the Subjective Visual Vertical Paradigm

Vestibular disorders are among the most common syndromes in medicine. In recent years, new vestibular diagnostic systems have been introduced that allow the examination of all semicircular canals in the clinical setting. Assessment methods of the otolithic system, which is responsible for the perception of linear acceleration and perception of gravity, are far less in clinical use. There are several experimental approaches for measuring the perception of gravity. The most frequently used method is the determination of the subjective visual vertical. This is usually measured with the head in an upright position. We present here an assessment method for testing otolith function in the roll plane. The subjective visual vertical is measured in the head upright position as well as with head inclination of &#177; 15&#176; and &#177; 30&#176; in the roll plane. This extended functional paradigm is an easy-to-perform clinical test of otolith function and ensures increased information content for the detection of impaired graviceptive perception.

The perception of gravity is commonly determined by the subjective visual vertical in the head upright position. The additional assessment at head tilts of &#177; 15&#176; and &#177; 30&#176; in the roll plane ensures increased information content for the detection of impaired graviceptive perception.

使用主观视觉垂直范式对卷平面中的静态视觉感知的评估

Vestibular disorders are among the most common syndromes in medicine. In recent years, new vestibular diagnostic systems have been introduced that allow ...

Validated LC-MS/MS Panel for Quantifying 11 Drug-Resistant TB Medications in Small Hair Samples

Drug resistant-tuberculosis (DR-TB) is a growing public health threat, and assessment of therapeutic drug levels may have important clinical benefits. Plasma drug levels are the current gold standard assessment, but require phlebotomy and a cold chain, and capture only very recent adherence. Our method uses hair, a matrix that is easily collected and reflective of long-term adherence, to test for 11 anti-TB medications. Previous work by our group shows that antiretroviral drug levels in hair are associated with HIV outcomes. Our method for DR-TB drugs uses 2 mg of hair (3 cm proximal to the root), which is pulverized and extracted in methanol. Samples are analyzed with a single LC-MS/MS method, quantifying 11 drugs in a 16 min run. Lower limits of quantification (LLOQs) for the 11 drugs range from 0.01 ng/mg to 1 ng/mg. Drug presence is confirmed by comparing ratios of two mass spectrometry transitions. Samples are quantified using the area ratio of the drug to the deuterated, 15N-, or 13C-labeled drug isotopologue. We used a calibration curve ranging from 0.001-100 ng/mg. Application of the method to a convenience sample of hair samples collected from DR-TB patients on directly observed therapy (DOT) indicated drug levels in hair within the linear dynamic range of nine of the eleven drugs (isoniazid, pyrazinamide, ethambutol, linezolid, levofloxacin, moxifloxacin, clofazimine, bedaquiline, pretomanid). No patient was on prothionamide, and the measured levels for ethionamide were close to its LLOQ (with further work instead examining the suitability of ethionamide&#8217;s metabolite for monitoring exposure). In summary, we describe the development of a multi-analyte panel for DR-TB drugs in hair as a technique for therapeutic drug monitoring during drug-resistant TB treatment.

Current methods of analyzing patients&#8217; adherence to complex drug resistant-tuberculosis (DR-TB) regimens can be inaccurate and resource-intensive. Our method analyzes hair, an easily collected and stored matrix, for concentrations of 11 DR-TB medications. Using LC-MS/MS, we can determine sub-nanogram drug levels that can be utilized to better understand drug adherence.

经过验证的LC-MS/MS面板，用于在小头发样本中量化11种耐药结核病药物

Drug resistant-tuberculosis (DR-TB) is a growing public health threat, and assessment of therapeutic drug levels may have important clinical benefits. ...

Using a Chemical Biopsy for Graft Quality Assessment

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated with an increased survival rate and greater quality of patient&#8217;s life when compared to conventional dialysis. Regrettably, transplantology suffers from a lack of reliable methods for organ quality assessment. Standard diagnostic techniques are limited to macroscopic appearance inspection or invasive tissue biopsy, which do not provide comprehensive information about the graft. The proposed protocol aims to introduce solid phase microextraction (SPME) as an ideal analytical method for comprehensive metabolomics and lipidomic analysis of all low molecular compounds present in kidneys allocated for transplantation. The small size of the SPME probe enables performance of a chemical biopsy, which enables extraction of metabolites directly from the organ without any tissue collection. The minimum invasiveness of the method permits execution of multiple analyses over time: directly after organ harvesting, during its preservation, and immediately after revascularization at the recipient&#8217;s body. It is hypothesized that the combination of this novel sampling method with a high-resolution mass spectrometer will allow for discrimination of a set of characteristic compounds that could serve as biological markers of graft quality and indicators of possible development of organ dysfunction.

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

使用化学活检进行移植质量评估

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...