Improper regulation of apoptosis has been postulated as one of the main factors that contributes to the etiology and/or progression of several prevalent diseases, including ischemic stroke and neurodegenerative pathologies. Consequently, in the last few years, there has been an ever-growing interest in the in vivo study of apoptosis. The clinical application of the tissue sampling and imaging approaches to analyze apoptosis in neurological diseases is, however, limited. Since apoptotic bodies are membrane vesicles that are released from fragmented apoptotic cells, it follows that the presence of these vesicles in the bloodstream is likely due to the apoptotic death of cells in tissues. We therefore propose to use circulating apoptotic bodies as biomarkers for measuring apoptotic death in patients with ischemic stroke and neurodegenerative diseases. Since there is no scientific literature establishing the most appropriate method for collecting and enumerating apoptotic bodies from human blood samples. Authors, here, describe a reproducible centrifugation-based method combined with flow cytometry analysis to isolate and quantify plasma apoptotic bodies of patients with ischemic stroke, multiple sclerosis, Parkinson’s disease and also in healthy controls. Electron microscopy, dynamic light scattering and proteomic characterization in combination with flow cytometry studies revealed that our isolation method achieves notable recovery rates of highly-purified intact apoptotic bodies. This easy, minimally time consuming and effective procedure for isolating and quantifying plasma apoptotic bodies could help physicians to implement the use of such vesicles as a non-invasive tool to monitor apoptosis in patients with cerebrovascular and neurodegenerative diseases for prognostic purposes and for monitoring disease activity.

中文翻译：

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血液凋亡小体的分离和量化，一种评估缺血性中风和神经退行性疾病患者细胞凋亡的非侵入性工具。

细胞凋亡的不当调节被认为是导致几种流行疾病（包括缺血性中风和神经退行性疾病）的病因和/或进展的主要因素之一。因此，在过去几年中，人们对细胞凋亡的体内研究越来越感兴趣。然而，组织取样和成像方法在分析神经疾病中细胞凋亡的临床应用是有限的。由于凋亡小体是从破碎的凋亡细胞中释放出来的膜囊泡，因此这些囊泡在血流中的存在很可能是由于组织中细胞的凋亡死亡。因此，我们建议使用循环凋亡小体作为生物标志物来测量缺血性中风和神经退行性疾病患者的凋亡死亡。由于没有科学文献确定从人体血液样本中收集和计数凋亡小体的最合适方法。作者在这里描述了一种可重复的基于离心的方法，结合流式细胞术分析，分离和量化缺血性中风、多发性硬化症、帕金森病和健康对照患者的血浆凋亡小体。电子显微镜、动态光散射和蛋白质组学表征结合流式细胞术研究表明，我们的分离方法实现了高度纯化的完整凋亡小体的显着回收率。这轻松，