Bu çalışma kısmen İsrail Kanser Araştırma Fonu&#39;ndan Ar-Ge Kariyer Geliştirme Ödülü, İsrail Ulusal Nanoteknoloji Girişimi ve Helmsley Bağışlı Güven&#39;den Kişiselleştirilmiş Teröristler için Nano Dedektifler (VP-K) için bir Odaklı Teknoloji Alanından hibe tarafından desteklendi ve Ulusal Sağlık yardımı enstitüleri R01GM076360 (XC&#39;ye).

<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. 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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>

Yazarların açıklayacak bir şeyi yok.

Başarılı bir glikan mikroarray üretimi dikkatli planlama gerektirir ve protokolde birkaç önemli adımı içerir. Bunlar arasında şunlar bulunur: (1) sonraki tüm parametreleri tanımlayan blok ve plaka düzenlerini planlama ( ör. Mesafeler, aralık, numune miktarı ve baskı); (2) piksellerin temizlenmesi ve nokta homojenliğini kontrol etmek için kritik olan pin bütünlüğünün sağlanması; (3) baskı esnasında yüksek nem oranının korunması, spot homojenliğinden ödün verebilecek uzun bas?...

Sias, hücre yüzeyi glikoproteinleri üzerindeki glıkan zincirlerini ve omurgalılardaki glikolipitleri kapsayan asidik şekerlerdir. Sia kanser hücrelerinde 1 değiştirilebilir ve ilerlemesi ve / veya metastaz 2, 3 ile bağıntılı olduğu ortaya konmaktadır. Memelilerdeki başlıca Sia formlarından ikisi Neu5Ac ve onun hidroksillenmiş formu Neu5Gc 2&#39;dir . İnsanlar CMAH enzimini kodlayan genin belirli bir inaktivasyonu nedeniyle Neu5Gc&#39;yi sentezleyemez. Bu insan dışı Sia metabolik diyet Neu5Gc zengin besinler (örneğin kırmızı et) den &quot;kendini,&quot; menşeli 4, 5 gibi insan hücrelerine sahiptir. Neu5Gc, insan epitelinin ve endotelin hücre yüzeylerinde düşük seviyelerde bulunur, ancak özellikle karsinomalarda birikir. Neu5Gc, insan humoral bağışıklık sistemi 2 , 6 tarafından yabancı olarak tanınır.Neu5Gc-glıkanların antijenik karmaşıklığı, insanlardaki anti-Neu5Gc antikor cevabının karmaşıklığı tarafından yansıtılan, Neu5Gc modifikasyonu, bağlanma, altta yatan glıkanlar ve iskeleler ve bunların yoğunluğu da dahil olmak üzere çoklu seviyelerde ortaya çıkabilir 6 . Bu antikorlardan bazıları karsinom biyolojik belirteçleri ve potansiyel immünoterapötikler 7 olarak kullanılır . Farklı sialoglikanlar 8&#39;in kemoenzimatik sentezinin ortaya çıkışı, bu tür antikorların daha derinlemesine analizine yol açtı, bu da glıkan mikroarray teknolojisi 9 , 10 kullanılarak kolaylaştırıldı. Bu nedenle, doğal ve sentetik karbonhidratların büyük kütüphanelerinin kolaylaştırılmış hazırlanması ve manipülasyonu ile, glıkan mikrodizileri karbonhidratların sayısız biyomoleküllerle olan etkileşimlerini araştırmak için güçlü, yüksek verimli bir teknoloji haline geldi 10 , 11 , </sup12 , 13 . Bir dizi formatında, az miktarda malzeme kullanılır ve biyolojik olarak ilgili glıkanların bu çok-değerli gösterimi, tek bir deneyde binlerce bağlanma etkileşiminin araştırılmasını sağlar. Önemli olan, bu teknoloji aynı zamanda biyomarkır bulguları ve çeşitli numunelerde 7 , 12 bağışıklık yanıtlarını izlemek için de uygulanabilir. Başarılı glıkan mikrodizi imalatı, üç önemli hususu göz önüne alır: yazıcı robot tipi, glikan konjugasyon kimyası ve tespit optiği. Baskı aleti konusuna gelince, iki teknik mevcuttur: temaslı ve temassız yazıcılar. Temas baskıda, 1-48 çelik pim, glikan solüsyonu içeren çok iyi kaynak plakasına daldırılır ve cam sürgü yüzeyine doğrudan temas ederek işlevselleştirilmiş cam slaytlar üzerinde lekelenir. Çözelti miktarı deSlayta gömülmüş olan, slayt yüzeyinde kalıcı sürenin bir fonksiyonudur. Genellikle, numuneler önce kaygan yüzey üzerine basılmadan önce bir cam blok üzerine (homojen noktalere ulaşmak için) önceden lekelenir. Temassız yazıcılarda ( örn., Piezo elektronik yazıcı), glikanlar, kontrollü elektrik sinyalleri kullanılarak bir cam kılcalından basılır. Kontak baskıya kıyasla daha hassas baskı elde etmek için elektrik sinyali hassas bir şekilde kalibre edilebilir. Noktaların boyutu ve morfolojisi de nispeten daha homojen. Ek bir avantaj, numunenin baskıdan sonra kaynak plakasına geri döndürülmesidir. Bununla birlikte, piezo elektronik yazıcıların en büyük dezavantajı baskı ucu sınırlaması (4 veya 8) olup, kayma kararlılığı, sıcaklık, nem ve numunenin buharlaşmasına özel dikkat gerektiren çok uzun bir yazdırma süresi sağlar. Temassız inkjet yazıcı 14 daha büyük numune hacimleri gerektirir. <p clasS = &quot;jove_content&quot;&gt; Yazdırma yöntemleri için mevcut sınırlı seçeneklerin aksine, glikan konjugasyon kimyası, daha pek çok seçeneğe sahip daha karmaşık bir sorundur. Seçilen immobilizasyon kimyası, hem glikanlardaki aktif grupları hem de slayt yüzeyinin reaktivitesini açıklamak zorundadır. Belirli bir mikrodizinin yüzeyinde, sentetik olarak sentezlenen veya doğal olarak izole edilen glikanlar, hepsi aynı reaktif grubu gerektirir. Buna ek olarak, glikanlar saf ve homojen olmalı. Öte yandan, immobilizasyon yüzeyi ve kimyası tekrarlanabilirlik ve güvenilir bağlanma yoğunluğu sağlamalıdır. Çoklu immobilizasyon yöntemleri, kovalent veya kovalent olmayan (fiziksel absorpsiyon) ataşman 10 , 11 , 12 , 13 ile geliştirilmiştir . Baskın glikan mikroarray teknolojisi ile ilgili son derece detaylı bilgi almak içinD araştırmacı, bu mükemmel değerlendirmeleri 13 , 15 bakın . Önemlisi, bir Glikomik Deney (MIRAGE) inisiyatifi için gereken son Minimum Bilgi, numune hazırlama 16 için ve bu büyüyen alanda standartları iyileştirmek için glıkan mikrodizi analizleri 17&#39;den veri raporlama talimatlarını açıklar. Burada, 16 yuvalı bir formatta belirli bir kontaklı nano-yazıcı kullanarak sialoglikan mikrodizilerinin imalatı için ayrıntılı bir protokolü açıklıyoruz. Glikanlardan her biri, epoksi ile aktive edilen cam slaytlarına kovalent bağlantısına aracılık eden bir birincil amin içerirler. Ayrıca çeşitli insan sera örneklerini, antikorları ve Sia bağlayıcı bitki lektinlerini kullanarak bir slaytın gelişimini ve analizini açıklıyoruz. Siyaloglikan mikroarray tahlilleri, dizi imalatı, işlenmesi, geliştirilmesi ve analizi içeren birkaç ana aşamayı içerir. Dizi üretiminde armanın planlanması gerekirRay düzeni, glikanlar ve kaynak plakası hazırlanması, nano yazıcının programlanması ve slaytların basılması. Daha sonra, slaytlar işlenir, geliştirilir ve analiz edilir ( Şekil 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

Hücreler, kanserde yaygın şekilde değişen ve sialik asit (Sia) ifadesinde değişiklikler içeren karbonhidrat zincirleri (glikanlar) paketi ile kaplıdır. Bunlar, 9-karbon omurgasına sahip olan ve hücre yüzeylerinde omurgalı glıkanları kaplayan asidik şekerlerdir. Memelilerin başlıca Sia formlarından ikisi N- asetilnöraminik asit (Neu5Ac) ve onun hidroksillenmiş formu, N- glikolilneuraminik asit (Neu5Gc) &#39;dir. İnsanlar, sitidin 5&#39;monofosfat-Neu5Ac (CMP-Neu5Ac) hidroksilaz (CMAH) kodlayan genin inaktivasyonu nedeniyle endojen Neu5Gc üretemez. Yabancı Neu5Gc, kırmızı et ve sütten besin tüketimi yoluyla insan hücreleri tarafından edinilir ve ardından çoğunlukla karsinomalar üzerine biriken hücre yüzeyindeki çeşitli glıkanlarda görülür. Sonuç olarak, insanlar kanser ve diğer kronik enflamasyon aracılı hastalıklarda çeşitli rol oynayan ve potansiyel tanı ve terapötik ta olan dolaşımdaki anti-Neu5Gc antikorlarına sahiptirler.rgets. Burada, bu tür anti-Neu5Gc antikorlarını insan serumunda değerlendirmek için yüksek verimli bir sialoglikan mikroarray deneyi tarif ederiz. Neu5Gc içeren glikanlar ve bunların her biri bir çekirdek primer amin içeren eşleştirilmiş kontrol çiftleri (Neu5Ac içeren glikanlar), epoksi kaplı cam slaytlara kovalent olarak bağlanır. Baskı başına 896 diziye kadar çıkabilen özel bir nano-yazıcı kullanarak 16 slayt formatında 56 slaytın baskısını örnekledik. Her slayt, anti-Neu5Gc antikor özgüllüğünün, yoğunluğunun ve çeşitliliğinin değerlendirilmesi için 16 farklı insan serum örneğini taramak için kullanılabilir. Protokol, bu sağlam aracın karmaşıklığını tanımlar ve bir dizi formatında farklı klinik örneklerde Neu5Gc diyet karbonhidrat antijenine cevabı araştırmayı amaçlayanların temel bir kılavuz sağlar.

Dizi Baskı, Geliştirme ve Analiz: Birden fazla glikan numunesi ve insan IgG STD eğrileri ile 16 farklı bloktaki bir sialoglikan mikroarrayi basmak, tüm numunelerin, slayt başına ve tüm slaytlara aynı baskıda 16 blokta mümkün olduğunca eşit şekilde basılmasını sağlamak için kapsamlı kalibrasyona ihtiyaç duyar. Bu nedenle, spesifik baskı parametreleri belirlenmeden önce, her bir numune türü için tampon ko...

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

Anti-Neu5Gc IgG&#39;yi İnsan Sera&#39;sında Sialoglikan Mikroarray Testi ile Görüntüleme

İnsan serasındaki anti-Neu5Gc antikorlarını değerlendirmek için bir siyaloglikan mikroarray testi kullanılabilir, böylece kanser ve diğer kronik enflamasyon aracılı insan hastalıkları için yüksek verimli bir tanı testi haline getirilir.

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

Research

JoVE Journal

Behavior

9.4K Görüntüleme.  Tel Aviv University.  İnsan serasındaki anti-Neu5Gc antikorlarını değerlendirmek için bir siyaloglikan mikroarray testi kullanılabilir, böylece kanser ve diğer kronik enflamasyon aracılı insan hastalıkları için yüksek verimli bir tanı testi haline getirilir. 

Video: 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.

Fonksiyonel Performans değerlendirilmesi<em&gt; Mdx</em&gt; Fare Modeli

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

Davranış

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.

Şizofreni ailesel Riskli Çocuklarda Duygusal Oddball Görev Kullanımı Fronto-limbik Faaliyet Ölçümü

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.

İntrakraniyal Elektrotlarla Bireylerde Davranış Görevler Sahne

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.

Yaşlı Erişkinlerde Nordic Walking Gait Değerlendirilmesi ve Postür için uygun Hareket Yakalama Teknikleri kanıtlayıcı

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.

Yüksek artı labirent takip yazılımı eksojen Ketogenic takviyeleri anksiyolitik etkisi araştırmak için Video ile kombine Test

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.

Cümle anlama Mandarin konuşan çocuklarda otizm eğitim için görsel dünya paradigması kullanarak

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.

Eğitimin Erken Evresinde Yüksek Yoğunluklu Aralıkegzersizve Orta Yoğunluklu Sürekli Egzersizin Kardiyak Troponin T Düzeyine Etkisi

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.

Bireysel Drosophila melanogaster Alkol Sedasyon Süresini Ölçmek için Yüksek İşLenme Yöntemi

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.

Yenilikçi Akıllı Telefon Tabanlı Diyet Değerlendirme Aracının Yinelemeli Gelişimi: 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.

Anne ve Bebeklerde Psikiyatrik Bozukluklar İçin Çevirisel olarak İlgili Bir Paradigma Olarak Gebelikte Psikososyal Stresin Murine Modelini Kullanmak

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