この研究は、イスラエル癌研究基金のResearch Career Development Award、Israeli National Nanotechnology Initiativeの助成金、Personalized Theranostics（VP-K）のNanomedicinesに関するFocal Technology AreaのHelmsley Charitable Trust、およびNational健康教育機関R01GM076360（XCへ）。

<ol>
	<li>Padler-Karavani, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aiming+at+the+sweet+side+of+cancer:+Aberrant+glycosylation+as+possible+target+for+personalized-medicine.">Aiming at the sweet side of cancer: Aberrant glycosylation as possible target for personalized-medicine.</a> Cancer Lett. 352 (1), 102-112 (2014).</li><li>Amon, R., Reuven, E. M., Leviatan Ben-Arye, ., Padler-Karavani, S., V, <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycans+in+immune+recognition+and+response.">Glycans in immune recognition and response.</a> Carbohydr Res. 389, 115-122 (2014).</li><li>H&#228;uselmann, I., Borsig, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+tumor-cell+glycosylation+promotes+metastasis.">Altered tumor-cell glycosylation promotes metastasis.</a> Front Oncol. 4, (2014).</li><li>Tangvoranuntakul, P., 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=Human+uptake+and+incorporation+of+an+immunogenic+nonhuman+dietary+sialic+acid.">Human uptake and incorporation of an immunogenic nonhuman dietary sialic acid.</a> Proc Natl Acad Sci USA. 100 (21), 12045-12050 (2003).</li><li>Bardor, M., Nguyen, D. H., Diaz, S., Varki, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mechanism+of+uptake+and+incorporation+of+the+non-human+sialic+acid+N-glycolylneuraminic+acid+into+human+cells.">Mechanism of uptake and incorporation of the non-human sialic acid N-glycolylneuraminic acid into human cells.</a> J Biol Chem. 280 (6), 4228-4237 (2005).</li><li>Padler-Karavani, V., 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=Diversity+in+specificity,+abundance,+and+composition+of+anti-Neu5Gc+antibodies+in+normal+humans:+potential+implications+for+disease.">Diversity in specificity, abundance, and composition of anti-Neu5Gc antibodies in normal humans: potential implications for disease.</a> Glycobiology. 18 (10), 818-830 (2008).</li><li>Padler-Karavani, V., 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=Human+xeno-autoantibodies+against+a+non-human+sialic+acid+serve+as+novel+serum+biomarkers+and+immunotherapeutics+in+cancer.">Human xeno-autoantibodies against a non-human sialic acid serve as novel serum biomarkers and immunotherapeutics in cancer.</a> Cancer Res. 71 (9), 3352-3363 (2011).</li><li>Cao, H., Chen, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=General+consideration+on+sialic+acid+chemistry.">General consideration on sialic acid chemistry.</a> Methods Mol Biol. 808, 31-56 (2012).</li><li>Deng, L., Chen, X., Varki, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exploration+of+sialic+Acid+diversity+and+biology+using+sialoglycan+microarrays.">Exploration of sialic Acid diversity and biology using sialoglycan microarrays.</a> Biopolymers. 99 (10), 650-665 (2013).</li><li>Liang, C. H., Hsu, C. H., Wu, C. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sialoside+Arrays:+New+Synthetic+Strategies+and+Applications.">Sialoside Arrays: New Synthetic Strategies and Applications.</a> Top Curr Chem. 367, 125-149 (2015).</li><li>Song, X., Heimburg-Molinaro, J., Smith, D. F., Cummings, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycan+microarrays+of+fluorescently-tagged+natural+glycans.">Glycan microarrays of fluorescently-tagged natural glycans.</a> Glycoconj J. 32 (7), 465-473 (2015).</li><li>Muthana, S. M., Gildersleeve, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycan+microarrays:+powerful+tools+for+biomarker+discovery.">Glycan microarrays: powerful tools for biomarker discovery.</a> Cancer Biomark. 14 (1), 29-41 (2014).</li><li>Rillahan, C. D., Paulson, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Glycan+microarrays+for+decoding+the+glycome.">Glycan microarrays for decoding the glycome.</a> Annu Rev Biochem. 80, 797-823 (2011).</li><li>Heimburg-Molinaro, J., Song, X., Smith, D. F., Cummings, R. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+and+analysis+of+glycan+microarrays.">Preparation and analysis of glycan microarrays.</a> Curr Protoc Protein Sci. 12 (10), (2011).</li><li>Park, S., Gildersleeve, J. C., Blixt, O., Shin, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Carbohydrate+microarrays.">Carbohydrate microarrays.</a> Chem Soc Rev. 42 (10), 4310-4326 (2013).</li><li>Struwe, W. B., 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=The+minimum+information+required+for+a+glycomics+experiment+(MIRAGE)+project:+sample+preparation+guidelines+for+reliable+reporting+of+glycomics+datasets.">The minimum information required for a glycomics experiment (MIRAGE) project: sample preparation guidelines for reliable reporting of glycomics datasets.</a> Glycobiology. 26 (9), 907-910 (2016).</li><li>Liu, Y., 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=The+minimum+information+required+for+a+glycomics+experiment+(MIRAGE)+project:+improving+the+standards+for+reporting+glycan+microarray-based+data.">The minimum information required for a glycomics experiment (MIRAGE) project: improving the standards for reporting glycan microarray-based data.</a> Glycobiology. , (2016).</li><li>Hara, S., Yamaguchi, M., Takemori, Y., Furuhata, K., Ogura, H., Nakamura, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+mono-O-acetylated+N-acetylneuraminic+acids+in+human+and+rat+sera+by+fluorometric+high-performance+liquid+chromatography.">Determination of mono-O-acetylated N-acetylneuraminic acids in human and rat sera by fluorometric high-performance liquid chromatography.</a> Anal Biochem. 179 (1), 162-166 (1989).</li><li>Padler-Karavani, V., 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=Cross-comparison+of+protein+recognition+of+sialic+acid+diversity+on+two+novel+sialoglycan+microarrays.">Cross-comparison of protein recognition of sialic acid diversity on two novel sialoglycan microarrays.</a> J Biol Chem. 287 (27), 22593-22608 (2012).</li><li>Padler-Karavani, V., Varki, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+impact+of+the+non-human+sialic+acid+N-glycolylneuraminic+acid+on+transplant+rejection+risk.">Potential impact of the non-human sialic acid N-glycolylneuraminic acid on transplant rejection risk.</a> Xenotransplantation. 18 (1), 1-5 (2011).</li><li>Samraj, A. 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. M., L&#228;ubli, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sialic+acids+in+cancer+biology+and+immunity.">Sialic acids in cancer biology and immunity.</a> Glycobiology. 26 (2), 111-128 (2016).</li><li>Alisson-Silva, F., Kawanishi, K., Varki, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+risk+of+diseases+associated+with+red+meat+intake:+Analysis+of+current+theories+and+proposed+role+for+metabolic+incorporation+of+a+non-human+sialic+acid.">Human risk of diseases associated with red meat intake: Analysis of current theories and proposed role for metabolic incorporation of a non-human sialic acid.</a> Mol Aspects Med. 51, 16-30 (2016).</li><li>Pearce, O. M., 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=Inverse+hormesis+of+cancer+growth+mediated+by+narrow+ranges+of+tumor-directed+antibodies.">Inverse hormesis of cancer growth mediated by narrow ranges of tumor-directed antibodies.</a> Proc Natl Acad Sci USA. 111 (16), 5998-6003 (2014).</li><li>Pham, T., 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=Evidence+for+a+novel+human-specific+xeno-auto-antibody+response+against+vascular+endothelium.">Evidence for a novel human-specific xeno-auto-antibody response against vascular endothelium.</a> Blood. 114 (25), 5225-5235 (2009).</li><li>Reuven, E. M., 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=Characterization+of+immunogenic+Neu5Gc+in+bioprosthetic+heart+valves.">Characterization of immunogenic Neu5Gc in bioprosthetic heart valves.</a> Xenotransplantation. 23 (5), 381-392 (2016).</li><li>Ghaderi, D., Taylor, R. E., Padler-Karavani, V., Diaz, S., Varki, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Implications+of+the+presence+of+N-glycolylneuraminic+acid+in+recombinant+therapeutic+glycoproteins.">Implications of the presence of N-glycolylneuraminic acid in recombinant therapeutic glycoproteins.</a> Nat Biotechnol. 28 (8), 863-867 (2010).</li></ol>

著者は何も開示することはない。

成功したグリカンマイクロアレイ作製には注意深い計画が必要であり、プロトコルにいくつかの重要なステップが含まれています。これらは、（1）すべての後続のパラメータ（ 例えば、距離、間隔、サンプルの量、および印刷）を定義ブロックおよびプレートのレイアウトを計画します。 （2）ピンの清掃およびピンの完全性の確保。これはスポットの均質性を制御するために重要で...

Siasは、脊椎動物の細胞表面糖タンパク質および糖脂質上のグリカン鎖を覆う酸性糖である。 SIA発現は、癌細胞1に変更し、進行および/または転移2,3と相関されます。哺乳類における主要なシアル酸の形の二つはNeu5Acで、そのヒドロキシル形、Neu5Gcを2です。ヒトは、CMAH酵素をコードする遺伝子の特異的不活性化のためにNeu5Gcを合成することができない。この非人間シアル酸は、代謝、食事のNeu5Gcが豊富な食品（ 例えば、赤肉）4、5から「自己」発信されたものとして、人間の細胞に組み込まれています。 Neu5Gcは、ヒト上皮および内皮の細胞表面上に低レベルで存在するが、特に癌腫に蓄積する。 Neu5Gcは、ヒト体液性免疫系2,6によって異物として認識されます。Neu5Gcグリカンの抗原複雑さは、すべての人間6における抗のNeu5Gc抗体応答の複雑さによって反射された、のNeu5Gc修正、結合、基礎となるグリカン及び足場、およびそれらの濃度を含む複数のレベルで生じ得ます。これらの抗体の一部は、癌腫バイオマーカーおよび潜在的な免疫療法剤として役立つ7 。異なるsialoglycans 8の化学酵素合成の出現は、グリカンマイクロアレイ技術9、10の使用によって容易にそのような抗体のより詳細な分析のために道を開きました。このように、天然および合成炭水化物の大きなライブラリーの促進準備と操作で、グリカンマイクロアレイは、生体分子10、11の無数と炭水化物の相互作用を調査するための強力なハイスループット技術となっています</sup&gt; 12、13。アレイ形式では、最小限の量の材料が使用され、生物学的に関連するグリカンのこの多価ディスプレイは、単一の実験において数千の結合相互作用の調査を可能にする。重要なことは、この技術はまた、バイオマーカー探索へと様々なサンプル7、12で免疫応答をモニタリングに適用することができます。 成功したグリカンマイクロアレイ作製には、プリンターロボットタイプ、グリカンコンジュゲーション化学、および検出光学系という3つの重要な側面を考慮する必要があります。印刷装置の考察に関しては、接触型および非接触型の2つの技術が利用可能である。接触印刷では、1〜48個のスチールピンを、グリカン溶液を含むマルチウエル源プレートに浸漬し、ガラススライド表面に直接接触させることにより官能化ガラススライド上にスポットする。解の量deスライドに生きているのは、スライド表面上の持続時間の関数である。通常、試料はスライド表面に印刷される前にガラスブロック上に先ず予めスポットされ（均質なスポットに到達する）。非接触プリンタ（ 例えば、圧電式プリンタ）では、制御された電気信号を用いてグリカンをガラスキャピラリから印刷する。電気信号は、接触印刷に比べてより正確な印刷を達成するために細かく較正することができる。スポットのサイズおよび形態も比較的均一である。さらなる利点は、印刷後にサンプルをソースプレートに再循環させることである。それにもかかわらず、ピエゾ電子式プリンタの主な欠点は、印刷チップの制限（4または8）であり、スライドの安定性、温度、湿度およびサンプルの蒸発に特に注意する必要がある非常に長い印刷時間をもたらす。非接触型インクジェットプリンタは、より大きなサンプル体積14を必要とする。 <p class = &quot;jove_content&quot;&gt;印刷法の利用可能なオプションが限られているのに対して、グリカン結合化学はより複雑な選択肢であり、選択肢は数多くあります。選択された固定化化学は、グリカン上の活性基および摺動表面反応性の両方を考慮しなければならない。特定のマイクロアレイ表面上に固定化されるグリカンは、合成的に合成されるかまたは天然に単離されるかのいずれも、同一の反応基を必要とする。さらに、グリカンは純粋で均質である必要があります。一方、固定化表面および化学物質は、再現性および信頼性のある付着密度を提供すべきである。複数の固定化方法は、共有結合または非共有結合（物理吸着）アタッチメント10、11、12、13のいずれかを用いて開発されてきました。 Uninitiateのための印刷されたグリカンマイクロアレイ技術に関する詳細な情報についてはDの研究者は、これらの優れたレビューは13、15を参照してください。重要なのは、グライコミクス実験（MIRAGE）イニシアチブのための最近の最低限必要な情報は、サンプル調製16のためにと、この成長分野で基準を改善するために17を解析し、糖鎖マイクロアレイからのデータを報告するためのガイドラインを説明します。 ここでは、特定の接触ナノプリンターを用いて16ウェルフォーマットでシアログリカンマイクロアレイを作製するための詳細なプロトコールについて説明する。各グリカンは、エポキシ活性化ガラススライドへのそれらの共有結合を媒介する第一級アミンを有する。我々はまた、様々なヒト血清サンプル、抗体、およびSia結合性植物レクチンを用いた1つのスライドの開発および分析について記載する。 Sialoglycanマイクロアレイアッセイは、アレイ作製、プロセシング、展開、および分析を含むいくつかの主要なステップを含む。アレイの製造には、arグリカンおよびソースプレートの準備、ナノプリンターのプログラミング、およびスライドの印刷を行うことができます。続いて、スライドを処理し、開発し、分析する（ 図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

細胞は、癌で一般的に変化し、シアル酸（Sia）発現の変化を含む炭水化物鎖（グリカン）の外皮で覆われている。これらは、9炭素骨格を有し、細胞表面上の脊椎動物グリカンをキャップする酸性糖である。哺乳動物の主要な2つのSia型は、 N-アセチルノイラミン酸（Neu5Ac）およびそのヒドロキシル化形態であるN-グリコリルノイラミン酸（Neu5Gc）である。ヒトは、シチジン5 &#39;モノホスフェート-Neu5Ac（CMP-Neu5Ac）ヒドロキシラーゼ（CMAH）をコードする遺伝子の不活性化のために、内因性Neu5Gcを産生することができない。外部Neu5Gcは、赤肉および乳製品の食物摂取を通じてヒト細胞によって獲得され、その後、細胞表面上の多様なグリカンに現れ、主に癌腫に蓄積する。結果として、ヒトは、癌および他の慢性炎症媒介性疾患において多様な役割を果たす循環型抗Neu5Gc抗体を有し、潜在的な診断および治療の可能性になっているラッツ。ここでは、ヒト血清中のそのような抗Neu5Gc抗体を評価するためのハイスループットのシアログリカンマイクロアレイアッセイを記載する。 Neu5Gc含有グリカンおよびそれらの対応する対の対照（Neu5Ac含有グリカン）は、それぞれコア第一アミンを有し、エポキシ被覆ガラススライドに共有結合される。我々は、1プリントあたり最大896個のアレイを生成することができる特定のナノプリンタを用いて、16ウェルフォーマットで56枚のスライドを印刷することを例示する。各スライドを使用して、抗Neu5Gc抗体特異性、強度および多様性の評価のための16の異なるヒト血清サンプルをスクリーニングすることができる。プロトコルは、この堅牢なツールの複雑さを記述し、配列形式で多様な臨床サンプルでNeu5Gc食物炭水化物抗原への応答を調査することを目指しているもののための基本的なガイドラインを提供します。

アレイ印刷、開発、分析： 複数のグリカンサンプルおよびヒトIgG STD曲線を含む16個の異なるブロックにシアログリカンマイクロアレイを印刷するには、スライドごとに16個のブロックすべてと同じ印刷操作ですべてのスライドにできるだけ均一にすべてのサンプルを印刷するための徹底的な較正が必要です。したがって、特...

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

Sialoglycanマイクロアレイアッセイによるヒト血清中の抗Neu5Gc IgGのプロファイリング

シアログリカンマイクロアレイアッセイを用いてヒト血清中の抗Neu5Gc抗体を評価し、癌および他の慢性炎症媒介ヒト疾患のハイスループット診断アッセイを可能にする。

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Behavior

9.4K ビュー.  Tel Aviv University. シアログリカンマイクロアレイアッセイを用いてヒト血清中の抗Neu5Gc抗体を評価し、癌および他の慢性炎症媒介ヒト疾患のハイスループット診断アッセイを可能にする。 

ビデオ: 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.

における機能のパフォーマンスを評価する<em&gt; MDX</em&gt;マウスモデル

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

行動学

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.

統合失調症のための家族ハイリスク児の感情オドボール課題を用いて前頭辺縁系の活性の測定

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.

頭蓋内電極を有する被験者に行動タスクの実行

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.

高齢者でノルディックウォーキング歩行や姿勢の評価のための適切なモーションキャプチャ技術を実証

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.

高架プラス迷路外因性ケト原性サプリメントの抗不安薬の効果を調べるためのソフトウェアを追跡するビデオと組み合わせてテスト

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.

北京語を話す自閉症児の文理を勉強する視覚的な世界のパラダイムを使用してください。

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.

トレーニングの初期段階における高強度インターバル運動と中強度連続運動が心臓トロポニンTレベルに及ぼす影響

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.

個々のショウジョウバエメラノガスターのアルコール沈殿時間を測定するためのハイスループット方法

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.

革新的なスマートフォンベースの食事評価ツールの反復開発: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.

母親と乳児の精神疾患に対する翻訳的に関連するパラダイムとしての妊娠における心理社会的ストレスのマウスモデルの使用

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