Bu çalışma Simons Vakfı (SFARI # 89638 RB) bir hibe ile desteklenmiştir.

Aşağıdaki protokol Washington Üniversitesi kurumsal inceleme kurulu kurallara bağlıdır. 1.. Elektrofizyolojik Değerlendirmesi <ol><li> Oturum hazırlanması <ol><li> Oda hazırlığı: place manipulandum (bkz. Şekil 1), bu katılımcının ulaşmak açgözlü içinde masaya, kavradı zaman toplama yazılımı için zaman damgalı bir işaretleyici gönderir bağlı bir sensör, bir ahşap blok. EEG toplama yazılımı etkinleştirin ve &quot;yeni bir oturum&quot; (Şekil S1) başlar. </li><li> Net hazırlanması: 104 ° F, damıtılmış su (1 L), potasyum klorür (1 çorba kaşığı), ve bebek şampuanı (1 çay kaşığı) sıcak çözeltisi Isındı tuzlu su çözeltisi 128-elektrot yoğun-dizisi EEG sistemi bekletin. </li><li> Katılımcı hazırlığı: katılımcı video cam görünümünde rahatça yaklaşık 75 cm uyaran sunum monitörden ve tam oturduğundan emindönemi. Bul ve bir cilt işaretleyici ile katılımcının kafasına köşe işaretleyin. Nasion ve inion ile preauriculars arasındaki orta noktası arasındaki orta noktanın kavşak bularak köşe ölçün. </li><li> Net uygulama: vertex elektrot tepe işareti üzerinde doğrudan yerleştirilir şekilde katılımcının kafasına EEG kapağını yerleştirin. Empedanslarını kontrol edin ve empedansları eşiğin altında kullanımda EEG sistemine (Şekil S2) için uygun olduğundan emin olun. </li><li> Video bantla oturumu başlayacak. </li></ol></li><li> Kayıt kurulum: köşe elektrot Referans sinyal. 0.1 ve 100 Hz arasında analog filtre, sinyali yükseltmek ve / sn 500 örneklerinde dijitalize. </li><li> Uyaran sunum: gözlemlemek yürütmek ve dinlenme, Muthukumaraswamy ve arkadaşları 5 tarafından geliştirilen paradigma adapte: 3 koşulları ile mevcut katılımcı. <ol><li> Durumunu gözlemlemek: sessizce oturup izlemek için katılımcı bilgilendirinmanipulandum doyumsuz bir kişinin video. Her deneme 6 sn sürmelidir. Tam gözlenen kavramak tam 3 sn gerçekleşir sağlamak için zaman gözlemlemek denemeleri için önceden kaydedilmiş video. Görev sırasında katılımcının görsel dikkatini izlemek, ve post-işleme sırasında atılması ekrana onlar gitmeyen sırasında denemeler işaretleyin. </li><li> Koşulu Execute: sadece manipulandum sağ el aşağıda dinlenme ile sessizce oturup, bir önceden kaydedilmiş işitsel işaret işitme üzerine, gözlemlemek koşulu video klipten manipulandum kapmak taklit için katılımcı bilgilendirin. Her deneme 6 sn sürmelidir. Işitsel işaret koruyan bir işitsel takip prerecording tarafından tam 3 sn sunulan emin olun sürekli işaret ve inter-deneme aralığını yürütmek uğradı. Tam katılımcının kavramak oluşur zaman (Şekil S3) kaydetmek için manipulandum bir sensör yararlanın. </li><li> Dinlenme durum: quie oturmak için katılımcı bilgilendirintly gözleri açık ve pasif uyaran monitörde küçük bir crosshair gözlemlemek ile. 3 dakika dinlenme durum sırasında Tutanak sürekli EEG. </li><li> Koşulları gözlemlemek ve yürütmek hem de, durum başına kırk çalışmaların toplam on çalışmaların mevcut randomize blok. Manipulandum görüntü denemeleri arasında da dahil olmak üzere, gözlemlemek ve yürütmek bloklar boyunca ekranda kalır emin olun. Gözlemlemek ve yürütmek koşulları bitiminde dinlenme durumunu yönetme. </li></ol></li><li> Veri İşleme <ol><li> Veri toplama takiben, empedanslarını tekrar kontrol edin. Empedans seviyelere herhangi bir değişiklik unutmayın. Toplama yazılımı Kaydı bitirmek. </li><li> Post-processing: ortalamasına Rereference EEG sinyali. Her durumda (Şekil S4) için kırk 6-sec çalışmalarda içine Segment sürekli EEG verileri. </li><li> Otomatik artefakt algılama Davranış. Exceedin hızlı ortalama genlikleri belirleyerek hareket eserler için segmentler incelemek için otomatik algoritmalar kullanıng 200 mV, 100 mV aşan ortalama genlikleri diferansiyel ve belirli bir deneme (Şekil S5) karşısında sıfır varyans. </li><li> Görsel verileri inceleme ve kavramak jest ilgisiz herhangi bir hareket dışlayıcı ile kontamine yürütme durumda herhangi bir hareket artefakt ve tüm denemeler ile kontamine gözlem durumda tüm denemeler kaldırmak için oturumu video inceleme ile teyit ederek manuel eserdir algılama Davranış. Analizinden önemli objeyle denemeler dışla. Katılmayan olarak satın alma sırasında işaretlenir olan tüm denemeler atın. Incelemek ve analiz altındaki her tanı grubunda için deneme ret oranı unutmayın. </li></ol></li><li> Veri analizi <ol><li> Başına Muthukumaraswamy vd. 5 hem gözlemlemek (fotosel ile işaretlenmiş olarak) ve (manipulandum sensör ile işaretlenmiş gibi) yürütmek koşulları sonra kavramak ve 1 saniye önce veri 1 sn oluşan 2 sn dönemini içine bölüm temizlenir denemeleri. Segment cgeri kalanı durumda 2 sn dönemini eğildi. </li><li> Hızlı Fourier Dönüşümü (FFT) Her segmenti dönüştürmek. Muthukumaraswamy vd. 5 aşağıdaki istatistiksel analizler için standart C3 ve C4 pozisyonları (çevreleyen her yarımkürede sekiz elektrot bir küme seçmek ve Bernier ve ark. 3) (Şekil 2). Her bir durum için, güç spektrumları hesaplamak için dahil deneme karşısında gücünü ortalama. </li><li> 8-13 Hz aralığında dinlenme durumu sırasında ortalama gücüne göre motor eylemin icrası veya gözlem, ya esnasında ortalama güç inceleyerek mu zayıflama hesaplayın. Zayıflatma derecesini belirlemek için bu oranın günlüğünü kullanın. Not: Bir pozitif bir değer büyütme temsil ederken negatif bir değer, yürütme ya da gözlem sırasında zayıflama temsil eder. Bu metodoloji, bireyler arasında değişkenlik hesap içine alır, ve oran şeklinde ifade değerlerinin olmayan normallik. &#39;br /&gt; Not: Bu protokol Net İstasyonu yazılım sürümü 4.1 ile 128-elektrot yoğun-dizisi EEG sistemi kullanılarak geliştirilmiştir. Temel adımlar EEG sistemleri arasında benzer olmakla birlikte, toplama ve analiz protokolleri değişebilir. </li></ol></li></ol> 2.. Örnek Karakterizasyon <ol><li> Araştırma kayıtları, önceki katılımcı listeleri, ya da alan klinikler ve klinisyenler tavsiyeleri ile paradigma katılım için potansiyel hasta popülasyonu tespit. </li><li> Klinik yapı (örneğin Otizm Spektrum Bozukluğu) tanı kriterlerini karşılayan ve bu kafa travması, tümör, nöbet öyküsü ya da bozabilir antikonvülsant veya uyku hapı ilaç kullanımının varlığı gibi herhangi bir dışlayıcı kriterleri belirlemek olasılığı için ekran potansiyel katılımcılar elektrofizyolojik sinyal. </li><li> Altın standart tanı araçlarının (örneğin, Otizm Tanı Görüşmesi-Re kullanımı ile hasta popülasyonunun tanılama durumu onaylamakDenetlenen (ADI-R 11) ve Otizm Tanı Gözlem Programı-Jenerik (ADOS-G, 12) Tanısal ve İstatistiksel El Kitabı aşağıdaki uzman klinisyen tarafından yönetilen - 5. Basım (DSM-5) kriterleri 13. </li><li> Vb yaş, cinsiyet, bilişsel yetenek, gibi ilgi ilgili değişkenler, Bahisler kontrol numunesini tespit </li></ol>

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N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+correlates+of+action+observation+and+execution+in+14&#8208;month&#8208;old+infants:+An+event&#8208;related+EEG+desynchronization+study.">Neural correlates of action observation and execution in 14&#8208;month&#8208;old infants: An event&#8208;related EEG desynchronization study.</a> Dev. Sci. , 474-480 (2011).</li><li>Marshall, P. J., Meltzoff, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neural+mirroring+systems:+Exploring+the+EEG+mu+rhythm+in+human+infancy.">Neural mirroring systems: Exploring the EEG mu rhythm in human infancy.</a> Dev. Cogn. Neurosci. , 110-123 (2011).</li><li>Oberman, L., McCleery, J., Hubbard, E., Bernier, R., Pineda, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developmental+changes+in+mu+suppression+to+observed+actions+in+individuals+with+autism+spectrum+disorders.">Developmental changes in mu suppression to observed actions in individuals with autism spectrum disorders.</a> Soc. Cogn. Affective Neurosci. 8, 300-304 .</li></ol>

Yazarlar herhangi bir mali çıkarlarını beyan.

Klinik nüfusa mu ritim ve uygulamaya ilişkin elektrofizyolojik verilerin başarılı bir satın alma, işleme ve analiz gerektirir 1) EEG metodolojik araçları uygulaması, 2) dikkatli dışlayıcı algılama ve veri azaltma, mu ritim 3) doğru tanımlama, ve 4) klinik nüfus ve uygun kontrol grupları tanımlanması doğru olarak nitelendirilebilir. Uygun EEG metodoloji düzgün ve özenle entegre ekipman, uygun kap seçimi ve yerleştirme, sinyalin doğru amp...

Elektroensefalografi (EEG), beyin aktivitesini değerlendirmek ve kayıt etkili, verimli ve invaziv olmayan bir yöntemdir. Nöronlar beyinde ateş gibi, ortaya çıkan gerilim, güçlendirilmiş kaydedilir ve grafiksel olarak temsil edilebilir. EEG zamansal çözünürlüğü daha kısa beyin salınım şekillerindeki değişiklikler, hem de belirli uyaranlara beynin tepkisinin analizi analizine izin verir. Geri 19. yüzyıl sonundan kalma eski beyin görüntüleme tekniği, olmasına rağmen, EEG hala geniş çaplı uygulanabilirliği vardır. Fonksiyonel manyetik rezonans görüntüleme (fMRI) mükemmel mekansal çözünürlüğe sahip olsa da, nispeten zayıf zamansal çözünürlüğe sahiptir. Bu prosesler, beyinde gerçekleştiği de inanılmaz hızı belirli FMRI değerlendirmesinin önemli bir sınırlandırmayı temsil etmektedir. EEG potansiyel i sağlayarak, milisaniyelik düzeyde elektriksel beyin aktivitesini değerlendirmek yeteneğine sahiptirbeynin işleme aşamaları içine NSight. Gelişen teknolojileri de EEG uygulanabilirliğini genişletmiştir. Kayıt sistemlerinin yoğunluğunda bir artış uzamsal çözünürlük konusunda EEG bazı sınırlamaları hafifletmek, kaynak lokalizasyonu tekniklerinin geliştirilmesi için izin verdi. Ayrıca, modern sistemler gibi bebek ve klinik örneklerin 1-3,28-30 olarak önceden kullanılamaz popülasyonlarının değerlendirilmesi için izin, önemli ölçüde bireysel katılımcı set-up süresini azalttı. Mükemmel zamansal çözünürlük göz önüne alındığında, EEG, belirli davranışların, devletler, ya da dış uyaranlara ilgili nöral cevabı incelemek için kullanılabilir. Bu programı bir örneği insanlarda ayna nöron sistemi (ANS) değerlendirilmesidir. Ayna nöronlar aslında bir grup kanıtlayan, tek bir nöron kayıt 4 kullanarak maymunlarda tespit edildiyürütülmesi ve motor işlemlerden gözlem her iki cevap nöronlar. Beyindeki elektrotlar yerleştirme Bu doğrudan kayıt yöntemi nadiren insanlarda ve sadece korkunç klinik durumlarda kullanılmaktadır. EEG EEG mu ritim izleyerek MNS değerlendirmek için bir yöntem sağlamıştır. 8-12 Hz aralığında bu salınım paterni yürütme ve maymunlarda 5-7 görülen aktivasyon desenine benzer motor eylemler, gözlem tepki olarak EEG gücünü azaltmak için gösterilmiştir. Benzer şekilde, Transkranial Manyetik Stimülasyon (örneğin inferior frontal girus) aracılığıyla varsayılan MNS beyin bölgelerinin uyarılması EEG mu ritim 8 kaldıracağına ve EEG mu ritim bastırma ek destek sağlayarak, bireylerde 9 içinde farazi ayna nöron bölgelerinde fMRI gelen BOLD sinyalleri ile ilişkili olduğunu bu ritim endeksler, en azından kısmen, MNS faaliyet. EEG mu ritim Değerlendirilmesi ayna nöron hareket noninvaziv değerlendirilmesi için izin verdiİnsanlarda ivity. EEG beyin aktivitesini incelemek için benzersiz bir yöntem sağlar ve davranışsal tabanlı deneyleri ile kombine edildiğinde, klinik toplumlarda, taklit gibi, sosyal biliş yönlerini ortaya çıkması için güçlü bir araç olabilir. Ayrıca, bilişsel ve dil bozuklukları ile nüfus ile kullanım için EEG uygulanabilirliği diğer görüntüleme teknikleri ya da davranışsal paradigmalar az başarılı kullanabilir olabilir kimin için bireylerin yetenekleri içgörü sağlar. Açıklanan protokol teorik böyle Otizm Spektrum Bozukluğu olarak tipik ve atipik gelişme, bireylerde ayna nöron sistemi ile bağlantılı sosyal bilişsel fonksiyonlarını incelemek için bir yol sağlar.

<table border="0" cellpadding="0" cellspacing="0" width="994">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Geodesic EEG System</td>
			<td style="width:71px;">EGI</td>
			<td style="width:119px;">N/A</td>
			<td style="width:539px;">Any EEG system, not only EGI based systems, is applicable for the described study</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">MATLAB software</td>
			<td style="width:71px;">MATLAB</td>
			<td style="width:119px;">N/A</td>
			<td>Any mathematical, statistical software that can work with matrices is applicable</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Netstation software</td>
			<td style="width:71px;">EGI</td>
			<td style="width:119px;">N/A</td>
			<td>Any EEG acquisition software is applicable for the described study</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:267px;">Manipulandum</td>
			<td style="width:71px;">custom</td>
			<td style="width:119px;">N/A</td>
			<td>Any object that is co-registered with data acquisition software to signal a successful grasp</td>
		</tr>
	</tbody>
</table>

eeg mu rhythm in typical and atypical development

Elektroensefalografi (EEG), beyin aktivitesini değerlendirmek ve kayıt etkili, verimli ve invaziv olmayan bir yöntemdir. Mükemmel zamansal çözünürlük göz önüne alındığında, EEG, belirli davranışların, devletler, ya da dış uyaranlara ilgili nöral cevabı incelemek için kullanılabilir. Bu programı bir örnek EEG mu ritim incelenmesi yoluyla insanlarda ayna nöron sistemi (ANS) değerlendirilmesidir. Sadece bireysel yürütür, ya da, amaca yönelik eylemleri gözlemler sırasında EEG mu ritim, merkezi bir konumda bulunan elektrotlar kaydedilen 8-12 Hz frekans aralığında salınım aktivite baskılanır. Bu nedenle, bu MNS aktivitesini yansıtan önerilmiştir. Bu ayna nöron sistemi (ANS) disfonksiyon otizm spektrum bozukluğu (ASD) toplumsal açıklarında bir katkıda rol oynadığı teorize edilmiştir. MNS sonra noninvaziv faaliyetinin bir göstergesi olarak EEG mu ritim zayıflama kullanılarak klinik popülasyonlarda incelenebilir. Anlatılan protocol teorik gibi ASD gibi tipik ve atipik gelişme, bireylerde MNS bağlantılı sosyal bilişsel fonksiyonlarını incelemek için bir yol sağlar.

Tipik yetişkinler, çocuklar ve bebekler sürekli yürütme ve eylemlerin gözlem çeşitli genelinde paradigmaların ve uyaranlara 5, 14-30 hem sırasında mu ritim göstermiştir. Bu frekans bandında zayıflama sürekli olarak bu diğer kafa derisi bölgelerinde kaydedilir alfa gücünün azaltılması olmadığını gösteren merkezi bir elektrot (Şekil 3) arasında yerelleştirilmiştir. Aynı şekilde, hareket gözlem sırasında bu frekansta zayıflama gibi (Şekil 4),</stron...

Watch this Scientific Journal Video about EEG Mu Rhythm in Typical and Atypical Development at JoVE.com

EEG Mu Rhythm in Typical and Atypical Development

EEG mu ritim Değerlendirilmesi beyin aktivitesini incelemek için benzersiz bir yöntem sağlar ve davranışsal tabanlı deneyleri ile kombine edildiğinde, klinik toplumlarda, taklit gibi, sosyal biliş yönlerini ortaya çıkması için güçlü bir araç olabilir.

Tipik ve Atipik Geliştirme EEG Mu Ritim

Electroencephalography (EEG) is an effective, efficient, and noninvasive method of assessing and recording brain activity. Given the excellent temporal ...

eeg-mu-rhythm-in-typical-and-atypical-development

Research

JoVE Journal

Medicine

25.4K Görüntüleme.  University of Washington.  EEG mu ritim Değerlendirilmesi beyin aktivitesini incelemek için benzersiz bir yöntem sağlar ve davranışsal tabanlı deneyleri ile kombine edildiğinde, klinik toplumlarda, taklit gibi, sosyal biliş yönlerini ortaya çıkması için güçlü bir araç olabilir. 

Video: EEG Mu Rhythm in Typical and Atypical Development

Direct Pressure Monitoring Accurately Predicts Pulmonary Vein Occlusion During Cryoballoon Ablation

Cryoballoon ablation (CBA) is an established therapy for atrial fibrillation (AF). Pulmonary vein (PV) occlusion is essential for achieving antral contact and PV isolation and is typically assessed by contrast injection. We present a novel method of direct pressure monitoring for assessment of PV occlusion.
Transcatheter pressure is monitored during balloon advancement to the PV antrum. Pressure is recorded via a single pressure transducer connected to the inner lumen of the cryoballoon. Pressure curve characteristics are used to assess occlusion in conjunction with fluoroscopic or intracardiac echocardiography (ICE) guidance. PV occlusion is confirmed when loss of typical left atrial (LA) pressure waveform is observed with recordings of PA pressure characteristics (no A wave and rapid V wave upstroke). Complete pulmonary vein occlusion as assessed with this technique has been confirmed with concurrent contrast utilization during the initial testing of the technique and has been shown to be highly accurate and readily reproducible.
We evaluated the efficacy of this novel technique in 35 patients. A total of 128 veins were assessed for occlusion with the cryoballoon utilizing the pressure monitoring technique; occlusive pressure was demonstrated in 113 veins with resultant successful pulmonary vein isolation in 111 veins (98.2%). Occlusion was confirmed with subsequent contrast injection during the initial ten procedures, after which contrast utilization was rapidly reduced or eliminated given the highly accurate identification of occlusive pressure waveform with limited initial training.
Verification of PV occlusive pressure during CBA is a novel approach to assessing effective PV occlusion and it accurately predicts electrical isolation. Utilization of this method results in significant decrease in fluoroscopy time and volume of contrast.

Effective pulmonary vein isolation utilizing a cryoballoon depends on complete pulmonary vein occlusion. The point of occlusion can be effectively predicted by direct analysis of pulmonary vein pressure waveform analysis during balloon inflation using a simple and reproducible technique.

Doğrudan Basıncı İzleme Doğru Cryoballoon Ablasyon sırasında Pulmoner Ven Tıkanıklığı tahmin

Cryoballoon  ablation (CBA) is an established therapy for atrial fibrillation (AF).  Pulmonary vein (PV) occlusion is essential for achieving antral ...

Tıp

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio based cues and set within the context of a video game metaphor, users gather relevant spatial information regarding a building's layout. This allows the user to develop an accurate spatial cognitive map of a large-scale three-dimensional space that can be manipulated for the purposes of a real indoor navigation task. After game play, participants are then assessed on their ability to navigate within the target physical building represented in the game. Preliminary results suggest that early blind users were able to acquire relevant information regarding the spatial layout of a previously unfamiliar building as indexed by their performance on a series of navigation tasks. These tasks included path finding through the virtual and physical building, as well as a series of drop off tasks. We find that the immersive and highly interactive nature of the AbES software appears to greatly engage the blind user to actively explore the virtual environment. Applications of this approach may extend to larger populations of visually impaired individuals.

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind.

Ses tabanlı Sanal Oyun Ortamı Geliştirilmesi Kör Navigasyon Becerileri ile Yrd etmek

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Clinical Examination Protocol to Detect Atypical and Classical Scrapie in Sheep

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated prion protein by post mortem tests. Unless a random sample of the sheep or goat population is actively monitored for scrapie, identification of scrapie cases relies on the reporting of clinical suspects, which is dependent on the individual&#39;s familiarization with the disease and ability to recognize clinical signs associated with scrapie. Scrapie may not be considered in the differential diagnosis of neurological diseases in small ruminants, particularly in countries with low scrapie prevalence, or not recognized&#160;if it presents as nonpruritic form like atypical scrapie. To aid in the identification of clinical suspects, a short examination protocol is presented to assess the display of specific clinical signs associated with pruritic and nonpruritic forms of TSEs in sheep, which could also be applied to goats. This includes assessment of behavior, vision (by testing of the menace response), pruritus (by testing the response to scratching), and movement (with and without blindfolding). This may lead to a more detailed neurologic examination&#160;of reporting animals as scrapie suspects. It&#160;could also be used in experimental TSE studies of sheep or goats to evaluate disease progression or to identify clinical end-point.

A short clinical examination protocol is presented to aid in the detection of classical and atypical scrapie in sheep and goats.

Koyunlarda Atipik ve Klasik Sıyrık Tespit Etmek için Klinik Muayene Protokolü

The diagnosis of scrapie, a transmissible spongiform encephalopathy (TSEs) of sheep and goats, is currently based on the detection of disease-associated ...

Real-time Cytotoxicity Assays in Human Whole Blood

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with high-throughput cell positioning technology. The targeted tumor cell populations are first preprogrammed to immobilization into an array format, and labeled with green fluorescent cytosolic dyes. Following the cell array formation, antibody drugs are added in combination with human whole blood. Propidium iodide (PI) is then added to assess cell death. The cell array is analyzed with an automatic imaging system. While cytosolic dye labels the targeted tumor cell populations, PI labels the dead tumor cell populations. Thus, the percentage of target cancer cell killing can be quantified by calculating the number of surviving targeted cells to the number of dead targeted cells. With this method, researchers are able to access time-dependent and dose-dependent cell cytotoxicity information. Remarkably, no hazardous radiochemicals are used. The WCA presented here has been tested with lymphoma, leukemia, and solid tumor cell lines. Therefore, WCA allows researchers to assess drug efficacy in a highly relevant ex&#160;vivo condition.

The whole blood cytotoxicity assay (WCA) is a cytotoxicity assay developed by incorporating high-throughput cell positioning technology with fluorescence microscopy and automated image processing. Here, we describe how lymphoma cells treated with an anti-CD20 antibody can be analyzed real-time in human whole blood to provide quantitative cellular cytotoxicity analysis.

İnsan Tüm Kan gerçek zamanlı sitotoksisite deneyleri

A live cell-based whole blood cytotoxicity assay (WCA) that allows access to temporal information of the overall cell cytotoxicity is developed with ...

Brain Source Imaging in Preclinical Rat Models of Focal Epilepsy using High-Resolution EEG Recordings

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with focal epilepsy. This methodology relies on the assumption that seizures originate from the same regions of the brain from which interictal epileptiform discharges (IEDs) emerge. Preclinical models are very useful to find correlates between IED locations and the actual regions underlying seizure initiation in focal epilepsy. Rats have been commonly used in preclinical studies of epilepsy1; hence, there exist a large variety of models for focal epilepsy in this particular species. However, it is challenging to record multichannel EEG and to perform brain source imaging in such a small animal. To overcome this issue, we combine a patented-technology to obtain 32-channel EEG recordings from rodents2 and an MRI probabilistic atlas for brain anatomical structures in Wistar rats to perform brain source imaging. In this video, we introduce the procedures to acquire multichannel EEG from Wistar rats with focal cortical dysplasia, and describe the steps both to define the volume conductor model from the MRI atlas and to uniquely determine the IEDs. Finally, we validate the whole methodology by obtaining brain source images of IEDs and compare them with those obtained at different time frames during the seizure onset.

This video introduces the preparation, recording, and source analysis procedures of high-resolution EEG on sedated rats with a particular preclinical model of focal epilepsy under noninvasive conditions.

Yüksek Çözünürlüklü EEG Kayıtlar kullanarak Odak Epilepsi Preklinik Rat Modelleri Beyin Kaynak Görüntüleme

Electroencephalogram (EEG) has been traditionally used to determine which brain regions are the most likely candidates for resection in patients with ...

Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody (mAb) by Neutralizing TNF Using an In Vitro Bioanalytical Method

This protocol shows the measurement of the apoptotic activity neutralization of TNF&#945; in a mouse fibroblast cell model (WEHI 164) using an anti-TNF&#945; mAb. In addition, this protocol can be used to evaluate other anti-TNF&#945; molecules, such as fusion proteins. The cellular model employed here is sensitive to TNF&#945;-mediated apoptosis when an additional stress factor is induced in cell culture conditions (e.g., serum deprivation). This procedure exemplifies how to execute this analytical assay, highlighting the key operations relating to the sample preparation, cell dilution, apoptosis induction, and spectrophotometric measurements that are critical to ensure successful results. This protocol reveals the best-performance conditions relating to apoptosis induction and efficient signal recording, leading to low uncertainty values.

Determination of the Relative Potency of an Anti-TNF Monoclonal Antibody mAb by Neutralizing TNF Using an In Vitro Bioanalytical Method

A protocol for the determination of the relative anti-apoptotic activity of an anti-TNF&#945; mAb using a neutralization mechanism with WEHI 164 cells is presented here. This protocol is useful for comparing the neutralization strength of different molecules with the same biological functionality.

Göreceli güç, bir Anti-TNF Monoklonal antikor (mAb) Neutralizing In Vitro Bioanalytical yöntemini kullanarak TNF tarafından belirlenmesi

This protocol shows the measurement of the apoptotic activity neutralization of TNF&#945; in a mouse fibroblast cell model (WEHI 164) using an ...

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&#946; and Tau Neuropathology

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. The paradigm includes tests for spontaneous alternation in a Y-maze, novel object recognition, and limb clasping. These tests were selected because they: 1) interrogate function of cognitive or motor domains and the correlate neural circuitry relevant to the human disease state, 2) have clearly defined endpoints, 3) have easily implementable quality control checks, 4) can be run in a moderate throughput format, and 5) require little intervention by the investigator. These methods are designed for investigators looking to screen compounds for activity in short-term and working memory tasks, or functional motor behaviors associated with Alzheimer's disease mouse models. The methods described here use behavioral tests that engage a number of different brain regions including hippocampus and various cortical areas. Investigators that desire cognitive tests that specifically assess cognition mediated by a single brain region could use these techniques to supplement other behavioral tests.

Assessment of Spontaneous Alternation, Novel Object Recognition and Limb Clasping in Transgenic Mouse Models of Amyloid-&amp;#946; and Tau Neuropathology

Described here is a staged, behavioral screening approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional motor behaviors in transgenic mouse models of &#946;-amyloidosis and tauopathy. These methods are optimized to screen compounds for activity in short-term and working memory tasks.

Amiloid-β ve Tau Nöropatolojisinin Transgenik Fare Modellerinde Spontan Dalgalanmanın Değerlendirilmesi, Yeni Nesne Tanıma ve Limb Klapması

Here we describe a staged, behavioral testing approach that can be used to screen for compounds that exhibit in vivo efficacy on cognitive and functional ...

The Pilocarpine Model of Temporal Lobe Epilepsy and EEG Monitoring Using Radiotelemetry System in Mice

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status epilepticus (SE) is frequently selected to recapitulate spontaneous recurrent seizures (SRS). Here we present a protocol of SE induction by intraperitoneal (i.p.) injection of pilocarpine and monitoring of chronic recurring seizures in live animals using a wireless telemetry video and electroencephalogram (EEG) system. We demonstrated notable behavioral changes that need attention after pilocarpine injection and their correlation with hippocampal neuronal loss at 7 days and 6 weeks post-pilocarpine. We also describe the experimental procedures of electrode implantation for video and EEG recording, and analysis of the frequency and duration of chronic recurrent seizures. Finally, we discuss the possible reasons why the expected results are not achieved in each case. This provides a basic overview of modeling chronic epilepsy in mice and guidelines for troubleshooting. We believe this protocol can serve as a baseline for suitable models of chronic epilepsy and epileptogenesis.

This manuscript describes a method of inducing status epilepticus by systemic pilocarpine injection and of monitoring spontaneous recurrent seizures in live animals using a wireless telemetry video and electroencephalogram system. This protocol can be utilized for studying the pathophysiologic mechanisms of chronic epilepsy, epileptogenesis, and acute seizures.

Temporal LOB epilepsi ve EEG farelerde Radiotelemetry sistemini kullanarak izleme Pilokarpin modeli

Temporal lobe epilepsy (TLE) is a common neurological disorder in adulthood. For translational studies of chronic epilepsy, pilocarpine-induced status ...

Contractions of Human-iPSC-derived Cardiomyocyte Syncytia Measured with a Ca-sensitive Fluorescent Dye in Temperature-controlled 384-well Plates

Spontaneously contracting syncytia of cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM) are a useful model of human cardiac physiology and pharmacology. Various methods have been proposed to record this spontaneous activity and to evaluate drug effects, but many of these methods suffer from limited throughput and/or physiological relevance. We developed a high-throughput screening system to quantify the effects of exogenous compounds on hiPSC-CM's beating frequency, using a Ca-sensitive fluorescent dye and a temperature-controlled imaging multi-well plate reader. We describe how to prepare the cell plates and the compound plates and how to run the automated assay to achieve high sensitivity and reproducibility. We also describe how to transform and analyze the fluorescence data to provide reliable measures of drug effects on spontaneous rhythm. This assay can be used in drug discovery programs to guide chemical optimization away from, or toward, compounds affecting human cardiac function.

Spontaneously contracting syncytia of cardiomyocytes derived from human-induced pluripotent stem cells are useful models of human cardiac physiology and pharmacology. Here we present a high-throughput screening system to quantify the effects of exogenous compounds on beating frequency, using a Ca-sensitive fluorescent dye and a temperature-controlled imaging multi-well plate reader.

384-şey plakaları ısı kontrollü bir Ca duyarlı floresan boya ile ölçülen insan IPSC türetilmiş Cardiomyocyte Syncytia kasılmalar

Spontaneously contracting syncytia of cardiomyocytes derived from human-induced pluripotent stem cells (hiPSC-CM) are a useful model of human cardiac ...

A Training and Testing System for Performing Vascular Reconstruction In Vitro

Manual vascular reconstruction training is essential for a beginner surgeon. However, an optimal training system for vascular reconstruction in vitro has yet to be developed. In this study, we introduce an in vitro training and testing system using a magnetic anchoring technique with which a trainee can practice manual vascular reconstruction individually. Additionally, this system can also be used to test the quality of the reconstruction. The described system includes a vascular reconstruction training machine, magnetic tractors, and a magnetic suture puller. In this manuscript, we detail an end-to-end vein anastomosis using porcine right and left iliac veins. To identify the potential damage caused by a magnetic suture puller on the suture, we created three groups with six segments of 4-0 polypropylene sutures each: a control group with no intervention on the polypropylene suture, a group in which the polypropylene suture is manually pulled with sterile gloves 20x, and a magnetic puller group in which the magnetic puller pulled the polypropylene suture 20x. These groups were tested by light microscopy and breaking strength tests, and the effect of reconstruction was assessed. In the light microscopy test, the control group was less likely to be damaged (p &#60; 0.05) and the number of damaged points of the manual group and magnetic puller group were similar (p &#62; 0.05). The results of the breaking strength test were compared across groups and no significant difference was observed (p &#62; 0.05). The end-to-end anastomosis of the porcine iliac veins was successfully performed using this training system, and the reconstructed veins could undergo 2.0 kPa perfusion pressure. Using this training and testing system the trainee can practice manual vascular reconstruction in vitro individually with the aid of magnetic tractors and a magnetic suture puller, and the quality of the reconstruction can be tested.

Here we present a training and testing system where a trainee can complete manual vascular reconstruction in vitro individually using a magnetic anchoring technique. The system can also be used to test the quality of reconstruction.

Vasküler RekonstrüksiyonUn Tüp Bebek Tespini Için Eğitim ve Test Sistemi

Manual vascular reconstruction training is essential for a beginner surgeon. However, an optimal training system for vascular reconstruction in vitro has ...