这项工作部分得到以色列癌症研究基金研究职业发展奖的支持，以色列国家纳米技术计划和赫尔姆斯利慈善信托基金会为个人化支持纳米医学（VP-K）和国家卫生署授予R01GM076360（至XC）。

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N., 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=A+red+meat-derived+glycan+promotes+inflammation+and+cancer+progression.">A red meat-derived glycan promotes inflammation and cancer progression.</a> Proc Natl Acad Sci USA. 112 (2), 542-547 (2015).</li><li>Pearce, O. 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作者没有什么可以披露的。

成功的聚糖微阵列制造需要仔细的规划，并包括协议中的几个重要步骤。这些包括:（1）规划定义所有后续参数（ 例如距离，间距，样品量和打印）的块和板布局; （2）清洁引脚并确保引脚的完整性，这对于控制点均匀性至关重要; （3）在印刷过程中保持高湿度，在长时间打印运行期间避免样品蒸发至关重要，这可能损害光斑均匀性; （4）选择正确的对齐和分析参数，这可以影响结果（ 例如，背景扣除方法，阈值和斑点尺寸灵活性）。 该方法可以进一步修改以满足特定的实验设计和目标。例如，可以修改预先识别号码以更好地适合要在阵列上打印的每种类型的材料。针清洗步骤期间可以优化打印以适应不同的印刷材料，具有较短或延长的洗涤周期。此外，可以修改针头每次浸渍的斑点数量，以包括更多或更少的斑点，但是需要对阵列上使用的每种类型的材料进行单独的校准。荧光标记物的类型及其浓度可以进行修饰，只要它含有合适的缀合化学基团（ 即用于环氧涂布载片的伯胺）即可。 STD曲线的浓度和类型可以延长（ 例如，人/小鼠/其他生物体IgG，IgM，IgA 等 ）。可优化缓冲条件以适应不同的材料，优选不含伯胺的材料以避免非特异性结合阵列，这将导致增加的背景。重要的是，为了最佳的检测Neu5Gc，生物阻断试剂必须不含Neu5Gc（ 例如避免BSA牛奶），因为这可能会减少Neu5Gc-唾液酸多糖检测，现象越来越背景19,20。应优化初级和次级检测浓度，以避免高背景和非特异性结合。此外，可以修改扫描参数（ 例如功率，增益和分辨率）以减少背景或增强低信号。最后，对齐和分析参数可以修改以适应高背景或不同形状的特征。关于报告聚糖微阵列数据的推荐标准的更多信息和指南最近由MIRAGE倡议17描述，最终可以促进数据共享和解释。 总之，聚糖微阵列提供了一种用于研究聚糖 - 生物分子相互作用的强大工具，可适用于各种生物样品。抗Neu5Gc抗体在人类疾病中起着不同的作用23 。例如，在癌症，抗Neu5Gc抗体扮演双重角色:一方面，他们作为潜在生物标志物，但在另一方面，他们作为潜在的治疗7，21，24。这些抗体也有助于动脉粥样硬化25的恶化，异种移植20,26的功效，和糖基化的生物治疗药物27的效果。因此，在各种人体样品中分析抗Neu5Gc抗体的兴趣越来越高，而高通量测定（如本文所述）可有助于进一步了解其在人类健康和疾病中的作用。

Sias是覆盖细胞表面糖蛋白上的聚糖链和脊椎动物中糖脂的酸性糖。 SIA表达在癌细胞1改性，并与进展和/或转移2,3相关。哺乳动物中主要的Sia形式的两种是Neu5Ac及其羟基化形式Neu5Gc 2 。由于编码CMAH酶的基因的特异性失活，人类不能合成Neu5Gc。这种非人的新航代谢合并到人体细胞的"自我"，从富含膳食中Neu5Gc的食物（ 如红肉）始发4,5。 Neu5Gc在人上皮细胞和内皮细胞表面的低水平存在，但特别积累在癌中。 Neu5Gc被人体液免疫系统2认为是外来的， 6 。Neu5Gc-聚糖的抗原性复杂性可能出现在多个水平，包括Neu5Gc修饰，连锁，底层聚糖和支架及其密度，都反映在人类6抗体Neu5Gc抗体的复杂性6 。一些抗体作为癌生物标志物和潜在的免疫治疗7。不同sialoglycans 8的酶法合成的到来铺平了道路这样的抗体的更深入的分析，通过使用聚糖微阵列技术9,10容易。因此，具有天然和合成碳水化合物的大文库的制备容易和操纵，聚糖微阵列已经成为一个强大的高通量技术用于研究与生物分子10，11的无数的碳水化合物的相互作用， </sup&gt; 12，13。以阵列形式，使用最少量的材料，并且生物相关聚糖的这种多重显示允许在单个实验中调查数千个结合相互作用。重要的是，这种技术也可以应用于生物标志物发现以及各种样品7中，12监测免疫应答。 成功的聚糖微阵列制造需要考虑三个重要方面:打印机器人类型，聚糖共轭化学和检测光学。关于印刷仪器的考虑，有两种技术可供选择:联系和非接触式打印机。在接触印刷中，将1-48个钢针浸入含有聚糖溶液的多孔源板中，并通过直接接触玻璃载片表面而在功能化玻璃载片上点样。溶液量活动到幻灯片是滑动表面上持续时间的函数。通常，样品首先在玻璃块上预先点样（达到同质点），然后再将其印刷在载玻片上。在非接触式打印机（ 例如，压电式打印机）中，使用受控电信号从玻璃毛细管印刷聚糖。可以对电信号进行精细校准，以实现相对于接触印刷的更精确的印刷。斑点的大小和形态也相对更均匀。另外的优点是在打印后将样品回收到源极板。然而，压电式打印机的主要缺点是打印头限制（4或8），导致打印时间非常长，需要特别注意滑动稳定性，温度，湿度和样品蒸发。非接触喷墨打印机需要更大的样品体积14 。 <p class ="jove_content"&gt;与印刷方法的有限可用选项相反，聚糖缀合化学是一个更复杂的考虑因素，有许多选择可供选择。选择的固定化学必须考虑聚糖上的活性基团和载玻片表面的反应性。要固定在特定微阵列表面上的合成或天然分离的聚糖都需要相同的反应性基团。此外，聚糖需要是纯的和均一的。另一方面，固定化表面和化学应提供重复性和可靠的附着密度。多种固定化方法已经开发了共价或非共价的（物理吸收）附件10，11，12，13。关于印刷聚糖微阵列技术的非常详细的信息ð调查，把这些优秀回顾13，15。重要的是，最近提供的一个糖尿病实验最低限度信息（MIRAGE）计划描述了样品制备的准则16，并报告了聚糖微阵列分析17的数据，以提高这一增长领域的标准。 在这里，我们描述了使用16孔格式的特定接触式纳米打印机制造sialoglycan微阵列的详细协议。每个聚糖具有介导它们与环氧基活化玻璃载片的共价键的伯胺。我们还描述了使用各种人血清样品，抗体和Sia结合植物凝集素的一个载玻片的开发和分析。 Sialoglycan微阵列测定涉及几个主要步骤，包括阵列制造，加工，开发和分析。阵列制造需要规划射线布局，制备聚糖和源极板，编程纳米打印机和打印幻灯片。随后，对载玻片进行处理，显影和分析（ 图1 ）。

<table border="0" cellpadding="0" cellspacing="0" width="1205">
	<tbody>
		<tr height="60">
			<td height="60" style="height:60px;width:269px;">Primary-amine containing sialoglycans</td>
			<td style="width:235px;">Glycohub, Inc., Davis, CA, USA (http://www.glycohub.com/services)</td>
			<td style="width:227px;">Contact info@glycohubusa.com for compound requests</td>
			<td style="width:476px;">Printed glycans</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:269px;">Monosodium phosphate monohydrate</td>
			<td style="width:235px;">Sigma</td>
			<td style="width:227px;">S9638</td>
			<td style="width:476px;">Printing buffer component</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Disodium phosphate heptahydrate</td>
			<td style="width:235px;">Sigma</td>
			<td style="width:227px;">S9390</td>
			<td style="width:476px;">Printing buffer component</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Phosphate buffered saline</td>
			<td style="width:235px;">Hy-Labs</td>
			<td style="width:227px;">BP-507/500D</td>
			<td style="width:476px;">Printing buffer/ incubation/washing buffer</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Tris-base</td>
			<td style="width:235px;">Sigma</td>
			<td style="width:227px;">T1503</td>
			<td style="width:476px;">Slide blocking reagent</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Glycerol</td>
			<td style="width:235px;">Sigma</td>
			<td style="width:227px;">G-7893</td>
			<td style="width:476px;">Printing buffer component</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Ethanolamine</td>
			<td style="width:235px;">Thermo-Fisher Scientific</td>
			<td style="width:227px;">0700/08</td>
			<td style="width:476px;">Slide blocking reagent</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Ovalbumin (Grade V)</td>
			<td style="width:235px;">Sigma</td>
			<td style="width:227px;">A5503</td>
			<td style="width:476px;">Slide Blocking protein</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Tween-20</td>
			<td style="width:235px;">Sigma</td>
			<td style="width:227px;">P7949</td>
			<td style="width:476px;">Slide washing detergent</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Alexa 555-Hydrazide</td>
			<td style="width:235px;">Thermo-Fisher Scientific</td>
			<td style="width:227px;">A20501MP</td>
			<td style="width:476px;">Marker on array</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;width:269px;">ChromPure Human IgG, whole molecule</td>
			<td style="width:235px;">Jackson Immunoresearch</td>
			<td style="width:227px;">009-000-003</td>
			<td style="width:476px;">Printing component</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Biotinylated- SNA</td>
			<td style="width:235px;">Vector Laboratories</td>
			<td style="width:227px;">B-1305</td>
			<td style="width:476px;">Plant Lectin &#8211; binding Sia-alpha2&#8211;6-linked</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Biotinylated-MALII</td>
			<td style="width:235px;">Vector Laboratories</td>
			<td style="width:227px;">B-1265</td>
			<td style="width:476px;">Plant Lectin &#8211; binding Sia-alpha2&#8211;3-linked</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Chicken-anti Neu5Gc IgY</td>
			<td style="width:235px;">BioLegend</td>
			<td style="width:227px;">146903</td>
			<td style="width:476px;">Primary detection</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Cy3-Streptavidin</td>
			<td style="width:235px;">Jackson Immunoresearch</td>
			<td style="width:227px;">016-160-0848</td>
			<td style="width:476px;">Biotin binding</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Cy3-anti Human IgG</td>
			<td style="width:235px;">Jackson Immunoresearch</td>
			<td style="width:227px;">109-165-088</td>
			<td style="width:476px;">Secondary detection against human IgG</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Cy3-anti Chicken IgY</td>
			<td style="width:235px;">Jackson Immunoresearch</td>
			<td style="width:227px;">703-165-155</td>
			<td style="width:476px;">Secondary detection against chicken IgY</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Human sera samples</td>
			<td style="width:235px;">Israeli Blood Bank</td>
			<td style="width:227px;"></td>
			<td style="width:476px;">Primary detection</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Compressed Nitrogen (Grade 5)</td>
			<td style="width:235px;"></td>
			<td style="width:227px;"></td>
			<td style="width:476px;">General dusting/drying tool</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Epoxy-coated slides</td>
			<td style="width:235px;">Corning</td>
			<td style="width:227px;">40044</td>
			<td style="width:476px;">Slides</td>
		</tr>
		<tr height="80">
			<td height="80" style="height:80px;width:269px;">Epoxy-coated slides</td>
			<td style="width:235px;">PolyAn</td>
			<td style="width:227px;">2D 104-00-221</td>
			<td style="width:476px;">Slides. In this type of slides the surface is more hydrophobic (compared to Coring slides) therefore the glycans Print Buffer would need to be supplemented with 0.005% Tween-20 to obtain 100 &#181;m size spots.</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">384-well microtiter plate</td>
			<td style="width:235px;">Genetix</td>
			<td style="width:227px;">2070</td>
			<td style="width:476px;">Printing plate</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">VWR lab marker</td>
			<td style="width:235px;">VWR</td>
			<td style="width:227px;">52877-310</td>
			<td style="width:476px;">Slide labeling</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Staining Tube</td>
			<td style="width:235px;">ArrayIt</td>
			<td style="width:227px;">MST</td>
			<td style="width:476px;">Slide developing tool</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Staining bath</td>
			<td style="width:235px;">VWR</td>
			<td style="width:227px;">25608-904</td>
			<td style="width:476px;">Slide developing tool</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Slides glass holders</td>
			<td style="width:235px;">VWR</td>
			<td style="width:227px;">631-9321</td>
			<td style="width:476px;">Slide developing tool</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">GenePix Scanner</td>
			<td style="width:235px;">Molecular devices</td>
			<td style="width:227px;">4000B</td>
			<td style="width:476px;">Slide scanner</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">LM-60 NanoPrinter</td>
			<td style="width:235px;">ArrayIt</td>
			<td style="width:227px;">LM-60</td>
			<td style="width:476px;">Array printer</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Pins</td>
			<td style="width:235px;">ArrayIt</td>
			<td style="width:227px;">946MP3</td>
			<td style="width:476px;">Printing pins</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">ProPlate Module</td>
			<td style="width:235px;">Grace Bio-Labs</td>
			<td style="width:227px;">P37004</td>
			<td style="width:476px;">Slide developing module</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:269px;">Distilled water</td>
			<td style="width:235px;">Bio-Lab</td>
			<td style="width:227px;">2321020500</td>
			<td style="width:476px;">Required for arrayer and humidifier</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;width:269px;">Electronic Multi Pippete, 8 Channel , volume range 2-125 &#956;L</td>
			<td style="width:235px;">Thermo-Fisher Scientific (Matrix)</td>
			<td style="width:227px;">MA-2131 Impact2 Equalizer 384</td>
			<td style="width:476px;">Multi pippete for sample dispansing into 384-well plate</td>
		</tr>
	</tbody>
</table>

profiling anti-neu5gc igg in human sera with a sialoglycan microarray assay

细胞覆盖着通常在癌症中被改变并包括唾液酸（Sia）表达变化的碳水化合物链（聚糖）的斗篷。这些是具有9碳骨架并且在细胞表面上覆盖脊椎动物聚糖的酸性糖。哺乳动物中主要的Sia形式中的两种是N-乙酰神经氨酸（Neu5Ac）及其羟基化形式， N-乙酰神经氨酸（Neu5Gc）。由于编码胞苷5&#39;-单磷酸-Neu5Ac（CMP-Neu5Ac）羟化酶（CMAH）的基因失活，人类不能产生内源性Neu5Gc。外源Neu5Gc由人类细胞通过红肉和乳制品的饮食消耗而获得，随后出现在细胞表面的多种聚糖上，主要集中在癌上。因此，人类具有循环的抗Neu5Gc抗体，其在癌症和其他慢性炎症介导的疾病中起着不同的作用，并且正在成为潜在的诊断和治疗rgets。在这里，我们描述了一种高通量sialoglycan微阵列测定法来评估人血清中的这种抗Neu5Gc抗体。含有Neu5Gc的聚糖及其配对的对照（含Neu5Ac的聚糖），每个具有核心伯胺，共价连接到环氧树脂涂布的玻璃片上。我们举例说明了使用特定的纳米打印机，可以以16孔格式打印56张幻灯片，每台打印机最多可生成896个阵列。每张幻灯片可用于筛选16种不同的人血清样品，以评估抗Neu5Gc抗体的特异性，强度和多样性。该协议描述了这种强大的工具的复杂性，并提供了一个基本准则，旨在探讨在阵列格式的不同临床样品中对Neu5Gc膳食碳水化合物抗原的反应。

阵列打印，开发和分析: 打印具有多个聚糖样品的sialoglycan微阵列和16个不同块中的人IgG STD曲线需要彻底校准，以确保所有样品在每个幻灯片所有16个块中以及在相同打印运行中的所有幻灯片尽可能均匀地打印。因此，在确定特定印刷参数之前需要进行多次校准实验，包括每种样品的缓冲液组合物，载玻片类型和制造，384孔板型，湿度水平，384孔板中的样品体积，预量的量每种类型样品的印刷，人IgG STD曲线浓度和标记类型。这种印刷的sialoglycan阵列载玻片的耐久性在室温下在黑暗中的真空密封箱中经过验证可持续长达10个月。在每个印刷实验中，均匀度是通过使用Sia结合凝集素和抗体的质量控制实验进一步监测和验证。通过比较来自相同打印运行的幻灯片之间和之间的样本，已经优化了开发和扫描协议以实现均匀性和准确性。使用16孔隔板开发载玻片，通过使用优化的封闭条件（化学和生物），洗涤和孵育时间，确保块之间的适当分离。主要和次要检测被广泛地校准，以确保最小的背景和非特异性结合的检测。在幻灯片扫描和分析时，将输出结果转移到电子表格文件并进行分析，以确定每个点的检测。如果不符合QC（如果％CV&gt; 20，斑点在四个重复的平均值之外的一个标准偏差的范围之外），则每个点都经过异常值检查并被忽略。然后，平均信号强度对于每个样品的重复斑点平均减去局部背景，以产生每个印刷样品的相对荧光单位（RFU）数据点。一旦IgG STD曲线的均匀性在所有测试的块中得到验证，RFU值被归一化为ng /μLIgG，允许在实验之内和之间更好地比较样品。 Sia结合高通量测定: Sialoglycan阵列可用于检测结合含Sia聚糖的决定簇（ 如蛋白质，凝集素，抗体，病毒等 ）。对于QC印刷后，SIA-结合植物凝集素和抗Neu5Gc的IgY被用于19。 SNA结合α2-6连接的Sia，MALII结合α2-3连接的Sia，抗Neu5Gc IgY结合含Neu5Gc的唾液酸，但不结合Neu5Ac含有sialoglycans 19 ， 如图2A所示。 QC实验的目的是通过使用四个不同的引脚来验证打印和识别的印刷块之间的差异。通过开发每个幻灯片的每个主要检测的四个块来监测块对块的变异性，并且通过将与四个引脚中的每一个打印的块与相同的主要检测部分进行比较来监控。然后进行比较以确保不存在重大差异。 人血清含有抗多样Neu5Gc抗体6含义多种人类疾病的2，21，22，23。为了评估人血清IgG中的sialoglycan识别，在印刷的唾液酸多糖微粒上分析来自健康人供体的12个血清使用荧光标记的抗人IgG（ 图2B ）发展并发展。每个印刷块含有人IgG STD曲线，其在所有方块中都是相当的和均匀的（ 图2C ）。只有在验证每个块中的STD曲线的质量（ 图2C ）之后，聚糖斑点结果根据块中的斜率进行归一化，然后乘以稀释因子。 如图2B所示，所有测试的人血清含有各种水平的抗Neu5Gc IgG，几乎没有识别出配对的含Neu5Ac的唾液酸对。此外，抗Neu5Gc IgG识别模式在这12种不同血清之间是高度多样的，无论是在强度和多样性水平上。 <img alt="图1" class="xfigimg" src="/files/ftp_upload/56094/56094fig1.jpg" /> 图1:概述阵列打印，开发和图像分析。 （ A ）Neu5Gc9Acα2-6GalNAcαProNH2显示为代表性的Neu5Gc含量的Sia聚糖，其中将伯胺印在环氧树脂涂覆的载玻片上。 （ B ）印刷阵列与各种Sia结合蛋白一起开发，随后用合适的荧光标记的二抗检测。每个子阵列可以单独开发。 （ C ）在荧光扫描仪中扫描探测幻灯片，生成使用扫描仪图像软件进一步分析的图像。子阵列覆盖着阵列上的每个斑点的网格，并为每个斑点检测到荧光。然后将结果转移到电子表格文件中。示意性地表示单个孔中的单个阵列。 <a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56094/56094fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <img alt="图2" class="xfigimg" src="/files/ftp_upload/56094/56094fig2v2.jpg" /> 图2:使用各种Sia结合蛋白的唾液酸多糖微阵列的谱图。 幻灯片与16个不同的主要检测部分开发，每个块中有一个。 （ A ）代表性的3块QC载片，显示Sia特异性植物凝集素SNA和MALII以及多克隆单特异性鸡抗Neu5Gc IgY的结合模式。结果以每个单独块（红色，白色和蓝色，分别表示第100 个 ， 第 50 个和第 0百分位数）的热图格式的RFU表示。 （ B ）以1:100稀释度测试12种不同的健康人血清，并用抗huamn IgG检测以描绘抗Neu5Gc IgG反应性。通过将每个值与每个特定bl中的IgG STD曲线斜率相除，将RFU数据归一化为ng /μL然后乘以稀释因子。对于所有组合的块（红色，白色和蓝色，分别代表第 100 个 ， 第 50 个和第 0 个百分位数），数据以热图格式表示。 （ C ）已经用于标准化数据的人血清开发的12个区块的人IgG STD曲线。 <a href="//ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56094/56094fig2v2large.jpg" target="_blank">请点击此处查看此图的较大版本。</a> <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 糖冠ID </td><td colspan="4"> 结构体 </td></tr><tr><td> 1 </td><td colspan="4"> Neu5,9Ac 2α3Galβ4GlcNAcβO（CH 2）2 CH 2 NH 2 </td></tr><tr><td> 2 </td><td colspan="4"> Neu5Gc9Acα3Galβ4GlcNAcβO（CH 2 ） 2 CH 2/ sub&gt; NH 2 </td></tr><tr><td> 3 </td><td colspan="4"> Neu5,9Ac 2α6Galβ4GlcNAcβO（CH 2）2 CH 2 NH 2 </td></tr><tr><td> 4 </td><td colspan="4"> Neu5Gc9Acα6Galβ4GlcNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td>五</td><td colspan="4"> Neu5Acα6GalNAcαO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 6 </td><td colspan="4"> Neu5Gcα6GalNAcαO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 7 </td><td colspan="4"> Neu5,9Ac 2α3Galβ3GlcNAcβO（CH 2）2 CH 2 NH 2 </td></tr><tr><td> 8 </td><td colspan="4"> Neu5Gc9Acα3Galβ3GlcNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 9 </td><td colspan="4"> Neu5,9Ac 2α3Galβ3GalNAcαO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 10 </td><td colspan="4"> Neu5Gc9Acα3Galβ3GalNAcαO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 11 </td><td colspan="4"> Neu5Acα3Galβ4GlcNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 12 </td><td colspan="4"> Neu5Gcα3Galβ4GlcNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 13 </td><td colspan="4"> Neu5Acα3Galβ3GlcNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 14 </td><td colspan="4"> Neu5Gcα3Galβ3GlcNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 15 </td><td colspan="4"> Neu5Acα3Galβ3GalNAcαO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 16 </td><td colspan ="4"&gt;Neu5Gcα3Galβ3GalNAcαO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 17 </td><td colspan="4"> Neu5Acα6Galβ4GlcNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 18 </td><td colspan="4"> Neu5Gcα6Galβ4GlcNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 19 </td><td colspan="4"> Neu5Acα6Galβ4GlcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 20 </td><td colspan="4"> Neu5Gcα6Galβ4GlcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 21 </td><td colspan="4"> Neu5Acα3Galβ4GlcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 22 </td><td colspan="4"> Neu5Gcα3Galβ4GlcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 23 </td><td colspan="4"> Neu5,9Ac 2α6GalNAcαO（CH 2）2 CH 2 NH 2 </td></tr><tr><td> 24 </td><td colspan="4"> Neu5Gc9Acα6GalNAcαO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 25 </td><td colspan="4"> Neu5Acα3GalβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 26 </td><td colspan="4"> Neu5Gcα3GalβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 27 </td><td colspan="4"> Neu5Acα6GalβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 28 </td><td colspan="4"> Neu5Gcα6GalβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 29 </td><td colspan="4"> Neu5,9Ac 2α3GalβO（CH 2）2 CH 2 NH 2 </td></tr><tr><td>三十</td><td colspan="4"&gt;Neu5Gc9Acα3GalβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 31 </td><td colspan="4"> Neu5,9Ac 2α6GalβO（CH 2）2 CH 2 NH 2 </td></tr><tr><td> 32 </td><td colspan="4"> Neu5Gc9Acα6GalβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 33 </td><td colspan="4"> Neu5Acα3Galβ3GalNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 34 </td><td colspan="4"> Neu5Gcα3Galβ3GalNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 35 </td><td colspan="4"> Neu5,9Ac 2α3Galβ3GalNAcβO（CH 2）2 CH 2 NH 2 </td></tr><tr><td> 36 </td><td colspan="4"> Neu5Gc9Acα3Galβ3GalNAcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td&gt; 37 </td><td colspan="4"> Neu5,9Ac 2α6Galβ4GlcβO（CH 2）2 CH 2 NH 2 </td></tr><tr><td> 38 </td><td colspan="4"> Neu5Gc9Ac6Galβ4GlcβO（CH 2 ） 2 CH 2 NH 2 </td></tr><tr><td> 39 </td><td colspan="4"> Neu5,9Ac 2α3Galβ4GlcβO（CH 2）2 CH 2 NH 2 </td></tr><tr><td> 40 </td><td colspan="4"> Neu5Gc9Ac3Galβ4GlcβO（CH 2 ） 2 CH 2 NH 2 </td></tr></tbody></table> 表1:印刷糖列表。 <table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody><tr><td> 小学/中学 </td><td> 抗体/凝集素 </td><td> 库存集中 </td><td> 特异性 </stroNG&gt; </td><td> 工作稀释/浓缩 </td></tr><tr><td rowspan="4">主要检测</td><td>生物素化MALII </td><td> 1 mg / mL </td><td>唾液酸α2-3键</td><td> 1:50，20μg/ mL </td></tr><tr><td>生物素化SNA </td><td> 2 mg / mL </td><td>唾液酸α2-6键</td><td> 1:100,20μg/ mL </td></tr><tr><td>鸡抗Neu5Gc IgY </td><td> ND </td><td> Neu5Gc唾液酸</td><td> 1:7.000 </td></tr><tr><td>人血清</td><td> 100％ </td><td>许多表位</td><td> 1:100 </td></tr><tr><td rowspan="3">二次检测</td><td>的Cy3-链霉</td><td> 0.75 mg / mL </td><td>生物素</td><td> 1:500,1.5μg/ mL </td></tr><tr><td> Cy3抗鸡IgY </td><td> 0.75 mg / mL </td><td>鸡IgY </td><td> 1:2,000,0.375μg/ mL </td></tr><tr><td> Cy3抗人IgG H + L </td><td> 0.6 mg / mL </td><td>呼玛n IgG </td><td> 1:1,500，0.4μg/ mL </td></tr></tbody></table> 表2:初级和次级检测蛋白的列表。 补充数据: <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56094/SUPP_040917Leviatan_Ben-Arye_JoVE_Rev1.docx">请点击此处下载此文件。</a>

Watch this Scientific Journal Video about Profiling Anti-Neu5Gc IgG in Human Sera with a Sialoglycan Microarray Assay at JoVE.com

Profiling Anti-Neu5Gc IgG in Human Sera with a Sialoglycan Microarray Assay

使用唾液酸多糖微阵列测定在人血清中分析抗Neu5Gc IgG

可以使用sialoglycan微阵列测定来评估人血清中的抗Neu5Gc抗体，使其成为癌症和其他慢性炎症介导的人类疾病的潜在的高通量诊断测定。

Cells are covered with a cloak of carbohydrate chains (glycans) that is commonly altered in cancer and that includes variations in sialic acid (Sia) ...

profiling-anti-neu5gc-igg-human-sera-with-sialoglycan-microarray

Nanyang Technological University

Research

JoVE Journal

Behavior

9.4K 次观看.  Tel Aviv University. 可以使用sialoglycan微阵列测定来评估人血清中的抗Neu5Gc抗体，使其成为癌症和其他慢性炎症介导的人类疾病的潜在的高通量诊断测定。 

视频: Profiling Anti-Neu5Gc IgG in Human Sera with a Sialoglycan Microarray Assay

Assessing Functional Performance in the Mdx Mouse Model

Duchenne muscular dystrophy (DMD) is a severe and progressive muscle wasting disorder for which no cure is available. Nevertheless, several potential pharmaceutical compounds and gene therapy approaches have progressed into clinical trials. With improvement in muscle function being the most important end point in these trials, a lot of emphasis has been placed on setting up reliable, reproducible, and easy to perform functional tests to pre clinically assess muscle function, strength, condition, and coordination in the mdx mouse model for DMD. Both invasive and noninvasive tests are available. Tests that do not exacerbate the disease can be used to determine the natural history of the disease and the effects of therapeutic interventions (e.g. forelimb grip strength test, two different hanging tests using either a wire or a grid and rotarod running). Alternatively, forced treadmill running can be used to enhance disease progression and/or assess protective effects of therapeutic interventions on disease pathology. We here describe how to perform these most commonly used functional tests in a reliable and reproducible manner. Using these protocols based on standard operating procedures enables comparison of data between different laboratories.

Assessing Functional Performance in the Mdx Mouse Model

The primary outcome measure in clinical trials for neuromuscular disorders is generally improved muscle function. Therefore, assessing the effect of potential therapeutic compounds on muscle performance pre clinically in mouse models is of great importance. We here describe several functional tests to address this.

评估在功能性能<em&gt; MDX</em&gt;小鼠模型

Duchenne muscular dystrophy (DMD) is a severe and progressive muscle wasting disorder for which no cure is available. Nevertheless, several potential ...

科研

行为学

Measurement of Fronto-limbic Activity Using an Emotional Oddball Task in Children with Familial High Risk for Schizophrenia

Adolescence is a critical developmental period where the early symptoms of schizophrenia frequently emerge. First-degree relatives of people with schizophrenia who are at familial high risk (FHR) may show similar cognitive and emotional changes. However, the neurological changes underlying the emergence of these symptoms remain unclear. This study sought to identify differences in frontal, striatal, and limbic regions in children and adolescents with FHR using functional magnetic resonance imaging. Groups of 21 children and adolescents at FHR and 21 healthy controls completed an emotional oddball task that relied on selective attention and the suppression of task-irrelevant emotional information. The standard oddball task was modified to include aversive and neutral distractors in order to examine potential group differences in both emotional and executive processing. This task was designed specifically to allow for children and adolescents to complete by keeping the difficulty and emotional image content age-appropriate. Furthermore, we demonstrate a technique for suitable fMRI registration for children and adolescent participants. This paradigm may also be applied in future studies to measure changes in neural activity in other populations with hypothesized developmental changes in executive and emotional processing.

This paper describes how to use the emotional oddball task and fMRI to measure brain activation in children and adolescents at familial high risk for schizophrenia (FHR).&#160;FMRI was used to measure differences in fronto-striato-limbic regions during an emotional oddball task. Children with FHR exhibited abnormal functional activation during adolescence.

的额边缘活度测量在儿童使用情绪穿着或行为古怪的任务与家族性高风险的精神分裂症

Adolescence is a critical developmental period where the early symptoms of schizophrenia frequently emerge. First-degree relatives of people with ...

Performing Behavioral Tasks in Subjects with Intracranial Electrodes

Patients having stereo-electroencephalography (SEEG) electrode, subdural grid or depth electrode implants have a multitude of electrodes implanted in different areas of their brain for the localization of their seizure focus and eloquent areas. After implantation, the patient must remain in the hospital until the pathological area of brain is found and possibly resected. During this time, these patients offer a unique opportunity to the research community because any number of behavioral paradigms can be performed to uncover the neural correlates that guide behavior. Here we present a method for recording brain activity from intracranial implants as subjects perform a behavioral task designed to assess decision-making and reward encoding. All electrophysiological data from the intracranial electrodes are recorded during the behavioral task, allowing for the examination of the many brain areas involved in a single function at time scales relevant to behavior. Moreover, and unlike animal studies, human patients can learn a wide variety of behavioral tasks quickly, allowing for the ability to perform more than one task in the same subject or for performing controls. Despite the many advantages of this technique for understanding human brain function, there are also methodological limitations that we discuss, including environmental factors, analgesic effects, time constraints and recordings from diseased tissue. This method may be easily implemented by any institution that performs intracranial assessments; providing the opportunity to directly examine human brain function during behavior.

Patients implanted with intracranial electrodes provide a unique opportunity to record neurological data from multiple areas of the brain while the patient performs behavioral tasks. Here, we present a method of recording from implanted patients that can be reproducible at other institutions with access to this patient population.

执行任务的行为与颅内电极主题

Patients having stereo-electroencephalography (SEEG) electrode, subdural grid or depth electrode implants have a multitude of electrodes implanted in ...

Substantiating Appropriate Motion Capture Techniques for the Assessment of Nordic Walking Gait and Posture in Older Adults

Nordic walking (NW) has become a safe and simple form of exercise in recent years, and in studying this gait pattern, various data collection techniques have been employed, each with positives and negatives. The aim was to determine the effect of NW on older adult gait and posture and to determine optimal use of different data collection systems in both short and long duration analysis. Gait and posture during NW and normal walking were assessed in 17 healthy older adults (age: 69 &#177; 7.3). Participants performed two trials of 6 Minute Walk Tests (6MWT) (1 with poles (WP) and 1 without poles (NP)) and 6 trials of a 5m walk (3 WP and 3 NP). Motion was recorded using two systems, a 6-sensor accelerometry system and an 8-camera 3-dimensional motion capture system, in order to quantify spatial-temporal, kinematic, and kinetic parameters.
With both systems, participants demonstrated increased stride length and double support and decreased gait speed and cadence WP compared to NP (p &#60;0.05). Also, with motion capture, larger single support time was found WP (p &#60;0.05). With 3-D capture, smaller hip power generation and moments of force were found at heel contact and pre-swing as well as smaller knee power absorption at heel contact, pre-swing, and terminal swing WP compared to NP, when assessed over one cycle (p &#60;0.05). Also, WP yielded smaller moments of force at heel contact and terminal swing along with larger moments at mid-stance of a gait cycle (p &#60;0.05). No changes were found for posture.
NW seems appropriate for promoting a normal gait pattern in older adults. Three-dimensional motion capture should primarily be used during short duration gait analysis (i.e. single gait cycle), while accelerometry systems should be primarily employed in instances requiring longer duration analysis such as during the 6MWT.

The aim was to substantiate optimal use of data collection techniques for Nordic walking gait and posture analysis. Three-dimensional motion capture should be used during short duration analysis (i.e. single gait cycle), while accelerometry should be employed for longer duration analysis (i.e. repeated cycles) like a 6 Minute Walk Test.

充实适当的运动捕捉技术的越野行走步态中老年人的评估和姿势

Nordic walking (NW) has become a safe and simple form of exercise in recent years, and in studying this gait pattern, various data collection techniques ...

Elevated Plus Maze Test Combined with Video Tracking Software to Investigate the Anxiolytic Effect of Exogenous Ketogenic Supplements

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The purpose of the method is to document the effect of various potential anxiolytic treatments on laboratory rodent models. The EPM test is based on the rodents&#39; proclivity toward protected, enclosed dark spaces and unconditioned fear of open spaces and heights, and their innate intense motivation to explore novel environments. The EPM test is a widely used behavioral test for investigating the anxiolytic or anxiogenic responses of rodents given drugs that are known to affect behavior. Observation demonstrating a decreased proportion of time spent on closed arms, an increased proportion of time spent on open arms, a reduced number of entries to closed arms, and an elevated number of entries to open arms measured by the EPM test may reflect reduced anxiety levels. Using this method, the effect of exogenous ketone supplements on anxiety-related behavior is tested in Sprague Dawley (SPD) rats. Exogenous ketone supplements are chronically fed to the rats for 83 days or subchronically and acutely orally gavaged, daily for 7 days, before conducting the EPM test. Behavioral data collection is performed using the SMART video tracking system by a blinded observer at the end of the treatments. The main findings indicate that the EPM test is an effective method to detect the ketone supplement-induced anxiolytic effect and can be considered a sensitive measure to assess changes in anxiety behavior associated with drug- or metabolic-based therapies.

Here, we present a protocol to investigate changes in the anxiety level of rodent animal models. The elevated plus maze (EPM) test, used together with a video tracking software, provides a reliable method to document the effect of various potential anxiolytic treatments in preclinical laboratory scenarios.

高架加迷宫试验结合视频跟踪软件研究外源性生酮类补充剂的毒性作用

The overall goal of this study is to describe the methodology of the elevated plus maze (EPM) test in combination with a video tracking software. The ...

Using the Visual World Paradigm to Study Sentence Comprehension in Mandarin-Speaking Children with Autism

Sentence comprehension relies on the ability to rapidly integrate different types of linguistic and non-linguistic information. However, there is currently a paucity of research exploring how preschool children with autism understand sentences using different types of cues. The mechanisms underlying sentence comprehension remains largely unclear. The present study presents a protocol to examine the sentence comprehension abilities of preschool children with autism. More specifically, a visual world paradigm of eye-tracking is used to explore the moment-to-moment sentence comprehension in the children. The paradigm has multiple advantages. First, it is sensitive to the time course of sentence comprehension and thus can provide rich information about how sentence comprehension unfolds over time. Second, it requires minimal task and communication demands, so it is ideal for testing children with autism. To further minimize the computational burden of children, the present study measures eye movements that arise as automatic responses to linguistic input rather than measuring eye movements that accompany conscious responses to spoken instructions.

We present a protocol to examine the use of morphological cues during real-time sentence comprehension by children with autism.

用视觉世界范式研究自闭症普通话儿童的句子理解

Sentence comprehension relies on the ability to rapidly integrate different types of linguistic and non-linguistic information. However, there is ...

Impact of High-intensity Interval Exercise and Moderate-Intensity Continuous Exercise on the Cardiac Troponin T Level at an Early Stage of Training

An elevation in cardiac troponin T (cTnT), as a highly specific biomarker of cardiomyocyte damage, after moderate-intensity continuous exercise (MCE) has been described. The exercise-induced cTnT response distorts the diagnostic role of the cTnT assay. Although high-intensity interval exercise (HIE) is growing in popularity and concerns remain about its safety, available data related to cTnT release after HIE is limited, which hampers the use of HIE as a health intervention. Here, we present three representative HIE protocols [traditional HIE (repeated 4 min cycling at 90% V&#775;O2max interspersed with 3 min rest, 200 kJ/session); sprint interval exercise (SIE, repeated 1 min cycling at 120% V&#775;O2max interspersed with 1.5 min rest, 200 kJ/session); and repeated sprint exercise (RSE, 40 x 6 s all-out sprints interspersed with 9 s rest)] and one representative MCE protocol (continuous cycling exercise at an intensity of 60% V&#775;O2max, 200 kJ/session). Forty-seven sedentary, overweight young women were randomly assigned to one of four groups (HIE, SIE, RSE, and MCE). Six bouts of respective exercise were performed by every single group, with each being 48 h apart. Meanwhile, for four groups, the duration of the entire testing period was identical, being 10 days. Before and after the first and final exercise bouts, an assessment was conducted of cTnT. The current study provides a frame of reference giving a clear picture of how a specific exercise session affects the circulating cTnT concentration at the early stage of training. The information may assist with clinical interpretations of post-exercise cTnT elevation and guide the prescription of exercise, especially for HIE.

Here, we present protocols of high-intensity interval and moderate-intensity continuous exercise to observe the response of circulating cardiac troponin T (cTnT) concentration to acute exercise over 10 days. The information may assist with clinical interpretations of post-exercise cTnT elevation and guide the prescription of exercise.

高强度间歇锻炼和中强度连续运动对心脏肌钙蛋白T级在训练早期阶段的影响

An elevation in cardiac troponin T (cTnT), as a highly specific biomarker of cardiomyocyte damage, after moderate-intensity continuous exercise (MCE) has ...

High-Throughput Method for Measuring Alcohol Sedation Time of Individual Drosophila melanogaster

Drosophila melanogaster provides an excellent model to study the genetic underpinnings of alcohol sensitivity. In contrast to studies in human populations, the Drosophila model allows strict control over genetic background, and virtually unlimited numbers of individuals of the same genotype can be reared rapidly under well-controlled environmental conditions without regulatory restrictions and at relatively low cost. Flies exposed to ethanol undergo physiological and behavioral changes that resemble human alcohol intoxication, including loss of postural control, sedation, and development of tolerance. Here, we describe a simple, low-cost, high-throughput assay for assessing alcohol sedation sensitivity in large numbers of single flies. The assay is based on video recording of single flies introduced without anesthesia in 24-well cell culture plates in a set-up that enables synchronous initiation of alcohol exposure. The system enables a single person to collect individual ethanol sedation data on as many as 2,000 flies within an 8 h work period. The assay can, in principle, be extended to assess the effects of exposure to any volatile substance and applied to measure effects of acute toxicity of volatiles on other insects, including other fly species.

High-Throughput Method for Measuring Alcohol Sedation Time of Individual Drosophila melanogaster

Current methods to measure alcohol sensitivity in Drosophila are designed to test groups of flies. We present a simple, low-cost, high-throughput assay for assessing alcohol sedation sensitivity in large numbers of single flies. The method does not require specialized tools and can be performed in any laboratory using common materials.

测量个体果蝇黑色素气器酒精镇分时间的高通量方法

Drosophila melanogaster provides an excellent model to study the genetic underpinnings of alcohol sensitivity. In contrast to studies in human ...

Iterative Development of an Innovative Smartphone-Based Dietary Assessment Tool: Traqq

To collect dietary intake data in a fast and reliable manner, a flexible and innovative smartphone application (app) called Traqq was developed (iOS/Android). This app can be used as a food record and&#160;24-h recall (or shorter recall periods). Different sampling schemes can be created on either prespecified or random days/times within a predetermined period for both methods, with&#160;push notifications to urge the participants to register their food intake. In case of non-response, notifications are automatically rescheduled to ensure complete data collection. For use as a food record, respondents can access the app and log their food intake throughout the day. Food records close automatically at the end of the day; recalls close after submission of the consumed items. The recall as well as the food record module provide access to an extensive food list based on the Dutch food composition database (FCDB), which can be accustomed to fit different research purposes. When selecting a food item, respondents are simultaneously prompted to insert portion size, i.e., in household measures (e.g., cups, spoons, glasses), standard portion sizes (e.g., small, medium, large), or weight in grams, and eating occasion/time of consumption. Portion size options can be adjusted, e.g., only entry in grams in case of a weighed food record or time of consumption instead of eating occasion). The app also includes a My Dishes function, which allows the respondent to create their own recipes or product combinations (e.g., a daily breakfast) and only report the total quantity consumed. Subsequently, the app accounts for yield and retention factors. The data are stored on a secure server. If desired, additional questions, i.e., in general or those related to specific food items or eating occasions can be incorporated. This paper describes the development of the system (app and backend), including expert evaluations and usability testing.

This article describes the protocol for the development of an innovative smartphone-based dietary assessment application Traqq, including expert evaluations and usability testing.

创新智能手机饮食评估工具的迭倍开发:Traqq

To collect dietary intake data in a fast and reliable manner, a flexible and innovative smartphone application (app) called Traqq was developed ...

Using a Murine Model of Psychosocial Stress in Pregnancy as a Translationally Relevant Paradigm for Psychiatric Disorders in Mothers and Infants

The peripartum period is considered a sensitive period where adverse maternal exposures can result in long-term negative consequences for both mother and offspring, including the development of neuropsychiatric disorders. Risk factors linked to the emergence of affective dysregulation in the maternal-infant dyad have been extensively studied. Exposure to psychosocial stress during pregnancy has consistently emerged as one of the strongest predictors. Several rodent models have been created to explore this association; however, these models rely on the use of physical stressors or a limited number of psychosocial stressors presented in a repetitive fashion, which do not accurately capture the type, intensity, and frequency of stressors experienced by women. To overcome these limitations, a chronic psychosocial stress (CGS) paradigm was generated that employs various psychosocial insults of different intensity presented in an unpredictable fashion. The manuscript describes this novel CGS paradigm where pregnant female mice, from gestational day 6.5 to 17.5, are exposed to various stressors during the day and overnight. Day stressors, two per day separated by a 2 h break, range from exposure to foreign objects or predator odor to frequent changes in bedding, removal of bedding, and cage tilting. Overnight stressors include continuous light exposure, changing cage mates, or wetting bedding. We have previously shown that exposure to CGS results in the development of maternal neuroendocrine and behavioral abnormalities, including increased stress reactivity, the emergence of fragmented maternal care patterns, anhedonia, and anxiety-related behaviors, core features of women suffering from perinatal mood and anxiety disorders. This CGS model, therefore, becomes a unique tool that can be used to elucidate molecular defects underlying maternal affective dysregulation, as well as trans-placental mechanisms that impact fetal neurodevelopment and result in negative long-term behavioral consequences in the offspring.

The chronic psychosocial stress (CGS) paradigm employs clinically relevant stressors during pregnancy in mice to model psychiatric disorders of mothers and infants. Here, we provide a step-by-step procedure of applying the CGS paradigm and downstream assessments to validate this model.

使用妊娠期心理社会压力的Murine模型作为母亲和婴儿精神障碍的翻译相关范式

The peripartum period is considered a sensitive period where adverse maternal exposures can result in long-term negative consequences for both mother and ...