| Using the Cecal Ligation and Puncture Model of Sepsis to Induce Rats to Multiple Organ Dysfunction
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Author:
Date:
2021-04-05
[Abstract] Sepsis is a dysregulated hyperinflammatory disease caused by infection. Sepsis leads to multiple organ dysfunction syndrome (MODS), which is associated with high rates of mortality. The cecal ligation and puncture (CLP) model has been widely used in animals and has become the gold-standard method of replicating features of sepsis in humans. Despite several studies and modified CLP protocols, there are still open questions regarding the multifactorial determinants of its reproducibility and medical significance. In our protocol, which is also aimed at mimicking the sepsis observed in clinical practice, male Wistar rats are submitted to CLP with adequate fluid resuscitation (0.15 M NaCl, 25 ml/kg BW i.p.) immediately after surgery. At 6 h after CLP, additional fluid therapy (0.15 M NaCl, 25 ...
[摘要] [摘要]败血症是由感染引起的失调的高炎症性疾病。败血症导致多器官功能障碍综合征（MODS），其与高死亡率相关。盲肠结扎穿刺（CLP）模型已在动物中广泛使用，并已成为在人类中复制败血症特征的金标准方法。尽管进行了多项研究并修改了CLP协议，但关于其可重复性和医学意义的多因素决定因素仍存在未解决的问题。在我们的协议中，其目的还在于模仿在临床实践中观察到的败血症，雄性Wistar大鼠在手术后立即接受CLP并进行足够的液体复苏（0.15 M NaCl，25 ml / kg BW ip）。CLP后6小时，进行额外的液体疗法（0.15 M NaCl，25 ml / kg BW sc）和用亚胺培南-西司他丁的抗生素疗法（单剂量14 mg / kg BW sc）。液体疗法和抗生素疗法的时机与患者进入重症监护室时的初始护理相对应。这种脓毒症模型为模拟人类脓毒症提供了有用的平台，并可能为开发新疗法奠定基础。
[背景]败血症的定义是宿主对感染的反应失调导致的危及生命的器官功能障碍（Singer等，2016）。它是全球主要的公共卫生问题，是多器官功能不全综合征（MODS）的主要原因，多器官功能不全综合征与医院内高死亡率相关（Torio and Moore ，201 6 ; Kellum et al。，2019）。1991年首次定义败血症时，重点是全身性炎症反应（ACCP / ...
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| Assessing Classical Olfactory Fear Conditioning by Behavioral Freezing in Mice
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Author:
Date:
2018-09-20
[Abstract] Classical fear conditioning typically involves pairing a discrete cue with a foot shock. Quantifying behavioral freezing to the learned cue is a crucial assay for neuroscience studies focused on learning and memory. Many paradigms utilize discrete stimuli such as tones; however, given mice are odor-driven animals and the wide variety of odorants commercially available, using odors as conditioned stimuli presents advantages for studies involving learning. Here, we describe detailed procedures for assembling systems for presenting discrete odor cues during single-day fear conditioning and subsequent analysis of freezing behavior to assess learning.
[摘要] 经典的恐惧条件反射通常涉及将离散的线索与脚部震动配对。 量化对学习线索的行为冻结是针对学习和记忆的神经科学研究的关键分析。 许多范例利用诸如音调之类的离散刺激; 然而，鉴于小鼠是气味驱动的动物和商业上可获得的各种气味剂，使用气味作为条件刺激对于涉及学习的研究具有优势。 在这里，我们描述了用于组装系统的详细程序，该系统用于在单日恐惧条件反射期间呈现离散的气味线索并随后分析冷冻行为以评估学习。
【背景】联想恐惧学习是几种焦虑症的根源，涉及将中性刺激与厌恶结果配对。这种配对产生强烈的行为恐惧反应，以冷冻（LeDoux，2003）的形式，对条件刺激，可以量化为恐惧学习和记忆的量度。离散刺激，如音调，通常被用作恐惧条件反射的条件刺激;然而，嗅觉提示在诱导学习冷冻方面也非常有效（Pavesi et al。，2012; Ross and Fletcher，2018）。这种联想恐惧学习方法不同于利用捕食者气味的方法，这种方法产生本能行为而非学习行为，使其成为快速评估嗅觉学习的理想选择。行为冻结，定义为缺乏所有自愿运动（Blanchard和Blanchard，1969; ...
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| A Microfluidic Device for Massively Parallel, Whole-lifespan Imaging of Single Fission Yeast Cells
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Author:
Date:
2018-04-05
[Abstract] Whole-lifespan single-cell analysis has greatly increased our understanding of fundamental cellular processes such as cellular aging. To observe individual cells across their entire lifespan, all progeny must be removed from the growth medium, typically via manual microdissection. However, manual microdissection is laborious, low-throughput, and incompatible with fluorescence microscopy. Here, we describe assembly and operation of the multiplexed-Fission Yeast Lifespan Microdissector (multFYLM), a high-throughput microfluidic device for rapidly acquiring single-cell whole-lifespan imaging. multFYLM captures approximately one thousand rod-shaped fission yeast cells from up to six different genetic backgrounds or treatment regimens. The immobilized cells are fluorescently imaged for over a ...
[摘要] 整个寿命的单细胞分析极大地增加了我们对细胞老化等基本细胞过程的理解。为了观察整个寿命期间的个体细胞，必须从生长培养基中移除所有后代，通常通过手动显微切割。然而，手动显微切割费力，低通量，并且与荧光显微镜不兼容。在这里，我们描述了多路复用裂变酵母寿命显微解剖器（multFYLM）的组装和操作，这是一种用于快速获取单细胞全寿命成像的高通量微流体装置。 multFYLM从多达六种不同的遗传背景或治疗方案中捕获约一千个杆状裂殖酵母细胞。将固定的细胞荧光成像超过一周，而将子代细胞从装置中取出。得到的数据集产生记录每个细胞复制寿命的高分辨率多通道图像。我们预计multFYLM将广泛适用于裂殖酵母（Schizosaccharomyces pombe）和其他对称分裂的单细胞生物的单细胞整个寿命研究。
【背景】细胞衰老导致细胞功能的累积下降，最终导致死亡。大多数关于细胞衰老的研究侧重于模型单细胞生物的复制寿命，例如出芽酵母酿酒酵母（Nyström和Liu，2014; Wasko和Kaeberlein，2014; Wierman和Smith，2014; Ruetenik和Barrientos ，2015）。细胞的复制寿命（RLS）被定义为母细胞在其生命过程中产生的女儿的数量（Henderson和Gottschling，2008; Sutphin等人，2014）。 ...
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