Motor Nöron Hastalığı Araştırma Avustralya Enstitüsü (MNDRIA), Sylvia ve Charles Viertel Vakfı Klinik Araştırmacı hibe, Ramaciotti Vakfı ve Ulusal Sağlık ve Avustralya Tıp Araştırma Konseyi (Proje hibe sayısı APP1024915) destek fon minnetle kabul edilmektedir.

1.. Hasta Hazırlama <ol><li> Prospektif ve ardışık hasta işe. Not: Aşağıdaki protokol Sydney Batı Bölgesi Sağlık Servisi İnsan Araştırmaları Etik Kurulları tarafından onaylanmıştır. Aydınlatılmış onam, tüm hastalar tarafından verildi. Hastalar Westmead Hastanesi&#39;nde ALS / nöromüsküler kliniği prospektif ve ardışık alındı. </li><li> Test için ALS hastalarının uygunluğunu belirler. ALS ya da bir nöromüsküler bozukluğu tanısı olmayan hastaların dışlanması. </li><li> Bir eşlik eden fokal nöropati, dirsek, bilek veya ulnar nöropati median nöropati hastaları dışlamak. </li><li> Örneğin diyabetik polinöropati gibi genelleştirilmiş nöropati hastaları, dışlamak. </li><li> Yaygın ALS veya nöromüsküler taklit bozuklukların tanısını doğrulamak için klinik takip hastaları araştırmak ve. </li><li> Bilgilendirilmiş onam al için ALS ve nöromüsküler mimik bozukluğu olan hastalarda dahil olmak üzere, tüm hastalar tarafından sağlanan emin olunl nörofizyolojik prosedürleri. </li></ol> 2. Klinik Değerlendirme <ol><li> Amyotrofik Lateral Skleroz Fonksiyonel Derecelendirme Ölçeği-Gözden Geçirilmiş (ALSFRS-R) 18 puan ile tüm ALS hastalarının klinik evreleme belirleyin. </li><li> Önce bildirilen formüle göre Tüm ALS hastalarında hastalığın ilerleme hızını belirlemek; Semptomların (48-ALSFRS-R) / süre) 19. </li><li> Tıbbi Araştırma Konseyi (MRC) skalası 20 kullanan tüm hastaların kas gücünü belirleyin. Üst uzuv oluşan toplam MRC skoru geliştirin (, dirsek fleksiyonu, omuz abduksiyon dirsek uzantısı; bileği dorsifeleksiyonu, parmak kaçırma, başparmak kaçırma, toplam puanı 60) ve alt ekstremite (kalça fleksiyonu, diz uzatma, ayak bileği, toplam puanı 30) kas grupları . Kas gücü normal ise toplam MRC skoru 90 olmalıdır. </li><li> Tüm ALS bulber veya uzuv olarak tanımlanan hastalığın başlangıç ​​site belirlemekhastalar. </li></ol> 3. Nörofizyolojik Değerlendirme <ol><li> Kaçıran kaydedilen bileşik kas aksiyon potansiyeli ile median ve ulnar sinirlerde (CMAP) tepkiler üzerine motor sinir iletim çalışmaları üstlenmek brevis (APB), birinci dorsal interosseous (DYY) ve kaçıran haneli minimi (ADM) kasları pollicis. </li><li> Bir alkol uygulaması takip cilt direncini azaltmak için bir aşındırıcı jeli, temizleme ile cilt yüzeyini temizleyerek, bilek uyarıcı siteleri hazırlayın. Maksimal yanıt CMAP (bu supramaksimal) üretimi için gerekli olan yoğunluğunun üzerinde% 20 uyarıcı akım ayarlayın. </li><li> Seçilen uyarım Alanı ses iletimi önlenir şekilde uyarılması için en düşük eşik değeri sergiler emin olun. </li><li> Bir alkol uygulanması, ardından deri direnci azaltmak için bir aşındırıcı jel, silme ile cilt yüzey temizleme her kası üzerinde kayıt yeri hazırlayın. </li><li> Bileşik motor acti boyutunu kaydedinAPB, ADM ve DYY kasların potansiyelinin (CMAP) üzerine. Her kasın üzerinde bir karın tendon düzende yerleştirilmiş 10 mm altın disk elektrodlar tarafından kaydedilen edilecek Cevapları. Referans elektrot başparmak (APB ve FDI CMAP kayıtları) ve basamak 5 bazın temel üzerinde konumlandırılmasını da Özellikle, ADM için (CMAP tepkisinin negatif kalkışı sağlanması ilgili kasın orta noktası üzerinde, aktif elektrot yerleştirmek CMAP kayıtları). </li><li> 5 cm ve 8 cm DYY&#39;yi mesafeye APB ve ADM bileşik motor aksiyon potansiyeli yanıtlar için uyarıcı katot ve aktif elektrot arasındaki mesafeyi ayarlayın. </li><li> Paraziti azaltmak için iletken jel ile, elektrot uyarıcı ve G1 arasında, elin dorsal elektro nötr topraklama plakası yerleştirin. Önceki nötr toprak uygulaması için bir alkol uygulanması, ardından deri direnci azaltmak için bir aşındırıcı jel, silme ile cilt yüzeyine temizleyerek sitesi hazırlamak. </li><li>Filtre ayarları 3 Hz (düşük frekans filtresi) ve 10 KHz (yüksek frekans filtresi) arasında vardır emin olun. </li><li> Süpürme hızı 20 msn veya 2 msn / bölümü için ayarlanmış olduğundan emin olun. </li><li> Kayıt CMAP yanıtları için duyarlılık 5 mV ayarlanmış olduğundan emin olun. </li><li> Çalışma boyunca stimülasyon yerinde sıcaklığı izlemek ve bacak sıcaklığı 32 ° C&#39;de muhafaza edilir sağlamak </li></ol> 4. Analiz ve Yorumlama <ol><li> ALS hastalarında APB, DYY ve ADM kas ve nöromüsküler hastalık mimik kontrolleri üzerinde bazal-tepe CMAP genlikleri (mV) ölçün. </li><li> Aşağıdaki gibi, APB ve DYY kaslar üzerinde kaydedilen CMAP çarpılması ve ADM kas içinde kaydedilen CMAP Bu ürünü bölerek bölünmüş el indeksi (Şekil 1) hesaplayın: SI = APB CMAP * DYY CMAP _________________________ &amp; #160; ADM CMAP </li><li> Tanısal doğruluk (STARD) kriterleri raporlama standartlarına uygun olarak ALS hastaları ve nöromüsküler mimik bozukluğu patolojik denetimler arasındaki SI değerleri karşılaştırarak bölünmüş el indeksi tanılama programı belirleyin. </li><li> Karakteristiği (ROC) eğrileri işletim alıcıyı kullanarak bölünmüş el indeksi için optimal tanı limit değerlerini belirleyin. ALS ve nöromüsküler bozukluğu hastalardan alınan SI değerleri için duyarlılık (y-ekseni) ve 1-özgüllüğü (x-ekseni) grafiğini çizerek ROC eğrisi türevi. </li></ol>

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Yazarlar ifşa hiçbir şey yok.

Bu çalışma ALS, yeni nörofizyolojik tanı biyomarkeri içinde bölünmüş el indeksi tanısal yarar bildiriyor. SI güvenilir 5.2 optimal tanı cut-off değeri ile, nöromüsküler bozukluklar ALS ayırt. SI azalma uzuv başlangıçlı ALS hastalarında en belirgin oldu. Önemlisi, yeni geliştirilen tanı kriterleri 11 başı olarak tanı &quot;olası&quot; kategorisinde sınıflandırıldı ALS hastalarının önemli bir kısmı, anormal SI sergiledi. Birlikte ele alındığında, bu çalışmada elde ...

Bölünmüş el, hipotenar kasların göreceli korunarak, iç el kaslarının tenar karmaşık bir grup [Abduktor pollisis brevis (APB) ve birinci dorsal interosseous (DYY)] tercihli atrofi anlamına gelir ve Amiotropik belirli ve erken klinik özelliktir lateral skleroz (ALS) 1-4. Bölünmüş el işareti ALS için özel olduğunu bulgu, ALS 3 tanısal bir belirteç olarak bölünmüş el işareti için potansiyel bir rol oynadığını düşündürmektedir. Bölünmüş el işaretinin Kantitasyonu, yeni bir biyolojik belirteç nörofizyolojik geliştirilmesi yoluyla, ALS tanısı yardım daha fazla olabilir. Özellikle, bölünmüş el fenomeni rakamlarla ayrılmış el indeksi (SI), bileşik kas aksiyon potansiyeli (CMAP) çarpılmasıyla elde edilir tenar kompleks kaslar (APB ve DYY) üzerinde kaydedilen ve CMAP Bu ürünü bölünmesi genlik HYP üzerinde kaydedilenothenar kaslar (yani kaçıran digiti minimide, ADM) 5. ALS tanısı 6 imzalar üst ve alt motor nöron bir arada kapsayan klinik tabanlı kriterlerine büyük ölçüde dayanır. Bu kriterler, ancak, özellikle de hastalık süreci 7-10 erken dönemlerinde ALS bir tanı konulmuş duyarsız kabul edildi. Tanı kriterlerinin bir yeni değişiklik 11 gelişmiş ve bu kriterler tanı duyarlılığını artırmak 12-16 görünmesine rağmen, artış hassasiyet bulber başlangıçlı ALS hastalarında 15 ile sınırlı görünüyor edildi. Patognomonik testi yokluğunda, ALS tanı önemli 8 gecikebilir. Sonuçta, klinik içine nöroprotektif tedaviler ve işe alım kurumu belki de kritik tedavi pencere dönemi 9, 17 ötesinde, gecikebilir. Sonuç olarak,SI tanılama yardımcı prospektif sporadik ALS hastalarında değerlendirildi.

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			<td height="20" style="height:20px;">Abrasive gel (Nu Prep)</td>
			<td>Weaver and Company</td>
			<td>N-TA\/P\H-3 ENG</td>
			<td>Skin preperation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td colspan="2">(www.doweaver.com)</td>
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			<td height="20" style="height:20px;">Alcohol wipe</td>
			<td>Triad Disposables</td>
			<td>103101</td>
			<td>Skin preperation</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td colspan="2">(www.triad-group.net)</td>
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			<td height="20" style="height:20px;">Recording gel (Ten20 Conductive gel)</td>
			<td>Weaver and Company</td>
			<td>N-TA\/P\H-3 ENG</td>
			<td>Recording motor responses</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td colspan="2">(www.doweaver.com)</td>
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			<td height="20" style="height:20px;">Amyotrophic lateral sclerosis rating scale-revised questionnaire</td>
			<td>Publication&#160;</td>
			<td>Experimental builder</td>
			<td>Stage disease</td>
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			<td colspan="2">(see Methods and References)</td>
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			<td height="20" style="height:20px;">Medical Research Council muscle strength scale</td>
			<td>Medical Research Council-UK</td>
			<td>Experimental builder</td>
			<td>Stage disease</td>
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			<td height="20" style="height:20px;"></td>
			<td colspan="2">(see Methods and References)</td>
			<td></td>
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			<td colspan="2" height="20" style="height:20px;">Table of equipment used</td>
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			<td height="20" style="height:20px;">Name of the equipment used</td>
			<td>Company</td>
			<td>Catalog number</td>
			<td>Comments</td>
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			<td></td>
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		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Nerve conduction machine (Synergy/Nicolet EDX)</td>
			<td>CareFusion (www.carefusion.com/medical-products/neurology)</td>
			<td>SA110106M</td>
			<td>To measure split hand index</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Synergy software</td>
			<td>CareFusion</td>
			<td>765654679</td>
			<td>To measure split hand index</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td colspan="3">(www.carefusion.com/medical-products/neurology)</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Tem millimeter gold cup disc electrodes</td>
			<td>Grass Technologies</td>
			<td>F-E5GH-60</td>
			<td>To record motor responses</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td colspan="2">(www.grasstechnologies.com)</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Neural earth plate</td>
			<td>Westmead Hospital</td>
			<td>Experimental builder</td>
			<td>To record limb temperature</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Thermometer</td>
			<td>Westmead Hospital</td>
			<td>Experimental builder</td>
			<td>To record limb temperature</td>
		</tr>
	</tbody>
</table>

utility of dissociated intrinsic hand muscle atrophy in the diagnosis of amyotrophic lateral sclerosis

Bölünmüş El olgu tenar kasların baskın israf anlamına gelir ve amiyotrofik lateral skleroz (ALS) erken ve özel bir özelliktir. Bir roman bölünmüş el indeksi (SI) bölünmüş el fenomeni ölçmek için geliştirilen, ve tanısal yarar ALS hastalarında değerlendirildi. Bölünmüş el indeksi bileşik kas aksiyon potansiyeli (CMAP) ürünü bölünmesi ile elde edilmiştir Abduktor digiti minimide kası üzerinde kaydedilen CMAP tarafından Abduktor pollisis brevis ve birinci dorsal interosseous kaslar üzerinde kaydedilen genlik. Bölünmüş el indeksi tanılama programını değerlendirmek amacıyla, ALS hasta prospektif olarak değerlendirildi ve sonuçları nöromüsküler bozukluğu hastalarında karşılaştırılmıştır. Nöromüsküler bozukluğu olan hastaların (p &lt;0.0001) göre ayrılmış el indeksi anlamlı ALS azaltıldı. Ekstremite-başlangıçlı ALS hastalar bölünmüş el endeksinde en büyük düşüş, ve bir değer sergiledidiğer nöromüsküler bozukluklar 5.2 ya da daha az güvenilir bir farklılaşmış ALS. Sonuç olarak, bölme el indeksi, belki de daha önceki bir tanı kolaylaştırmak, ALS için yeni bir tanısal biyolojik olarak görünmektedir.

Klinik Fenotip Toplamda, 44 hastada ALS% 76 (N = 33) mümkün ALS olarak kesin ya da muhtemel ve% 24 (N = 11) olarak sınıflandırıldı Awaji kriterlere 11 göre, bunlardan incelenmiştir. Kapsamlı araştırmalar ve 3 yıla kadar ve% 53 klinik takip, bu dönemde öldükten sonra ALS teşhisi &quot;olası&quot; grubunda teyit edildi. Bulber başlangıçlı hastalık% 41 belli oldu, ise ALS hastalarının% 59 ekstremite başlangıçlı. Değerlendirme zamanda, hastalık s?...

Watch this Scientific Journal Video about Utility of Dissociated Intrinsic Hand Muscle Atrophy in the Diagnosis of Amyotrophic Lateral Sclerosis at JoVE.com

Utility of Dissociated Intrinsic Hand Muscle Atrophy in the Diagnosis of Amyotrophic Lateral Sclerosis

Iç el kaslannda aynştınlmış atrofisi, bölünmüş el olarak adlandırılan, amyotrofik lateral skleroz (ALS), belirli bir özellik olarak görülmektedir. Sonuç olarak, bir roman Neurodiagnostic testi, mimik bozukluğu ALS ayırt bölünmüş elin klinik fenomen, ölçmek için geliştirilen, bölünmüş el endeksi olarak adlandırılır.

Amyotrofik Lateral Skleroz Tanısında Disosiye ıntrinsic El kas Atrofi Utility

The split hand phenomenon refers to predominant wasting of thenar muscles and is an early and specific feature of amyotrophic lateral sclerosis (ALS). A ...

utility-dissociated-intrinsic-hand-muscle-atrophy-diagnosis

Research

JoVE Journal

Medicine

31.2K Görüntüleme.  Westmead Hospital.  Iç el kaslannda aynştınlmış atrofisi, bölünmüş el olarak adlandırılan, amyotrofik lateral skleroz (ALS), belirli bir özellik olarak görülmektedir. Sonuç olarak, bir roman Neurodiagnostic testi, mimik bozukluğu ALS ayırt bölünmüş elin klinik fenomen, ölçmek için geliştirilen, bölünmüş el endeksi olarak adlandırılır. 

Video: Utility of Dissociated Intrinsic Hand Muscle Atrophy in the Diagnosis of Amyotrophic Lateral Sclerosis

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis (ALS)

Improved methods for assessing bulbar impairment are necessary for expediting diagnosis of bulbar dysfunction in ALS, for predicting disease progression across speech subsystems, and for addressing the critical need for sensitive outcome measures for ongoing experimental treatment trials. To address this need, we are obtaining longitudinal profiles of bulbar impairment in 100 individuals based on a comprehensive instrumentation-based assessment that yield objective measures. Using instrumental approaches to quantify speech-related behaviors is very important in a field that has primarily relied on subjective, auditory-perceptual forms of speech assessment1. Our assessment protocol measures performance across all of the speech subsystems, which include respiratory, phonatory (laryngeal), resonatory (velopharyngeal), and articulatory. The articulatory subsystem is divided into the facial components (jaw and lip), and the tongue. Prior research has suggested that each speech subsystem responds differently to neurological diseases such as ALS. The current protocol is designed to test the performance of each speech subsystem as independently from other subsystems as possible. The speech subsystems are evaluated in the context of more global changes to speech performance. These speech system level variables include speaking rate and intelligibility of speech.
The protocol requires specialized instrumentation, and commercial and custom software. The respiratory, phonatory, and resonatory subsystems are evaluated using pressure-flow (aerodynamic) and acoustic methods. The articulatory subsystem is assessed using 3D motion tracking techniques. The objective measures that are used to quantify bulbar impairment have been well established in the speech literature and show sensitivity to changes in bulbar function with disease progression. The result of the assessment is a comprehensive, across-subsystem performance profile for each participant. The profile, when compared to the same measures obtained from healthy controls, is used for diagnostic purposes. Currently, we are testing the sensitivity and specificity of these measures for diagnosis of ALS and for predicting the rate of disease progression. In the long term, the more refined endophenotype of bulbar ALS derived from this work is expected to strengthen future efforts to identify the genetic loci of ALS and improve diagnostic and treatment specificity of the disease as a whole. The objective assessment that is demonstrated in this video may be used to assess a broad range of speech motor impairments, including those related to stroke, traumatic brain injury, multiple sclerosis, and Parkinson disease.

A Protocol for Comprehensive Assessment of Bulbar Dysfunction in Amyotrophic Lateral Sclerosis ALS

Objective assessments of the physiological mechanisms that support speech are needed to monitor disease onset and progression in persons with ALS and to quantify treatment effects in clinical trials. In this video, we present a comprehensive, instrumentation-based protocol for quantifying speech motor performance in clinical populations.

Amyotrophic Lateral Sclerosis bulber Disfonksiyon Kapsamlı Değerlendirmesi (ALS) için Protokol

Improved methods for assessing bulbar impairment are necessary for expediting diagnosis of bulbar dysfunction in ALS, for predicting disease progression ...

Tıp

Intraspinal Cell Transplantation for Targeting Cervical Ventral Horn in Amyotrophic Lateral Sclerosis and Traumatic Spinal Cord Injury

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) cases 1 and is the ultimate cause of death in patients with the motor neuron disorder, amyotrophic laterals sclerosis (ALS) 2.
ALS is a devastating neurological disorder that is characterized by relatively rapid degeneration of upper and lower motor neurons. Patients ultimately succumb to the disease on average 2-5 years following diagnosis because of respiratory paralysis due to loss of phrenic motor neuron innnervation of the diaphragm 3. The vast majority of cases are sporadic, while 10% are of the familial form. Approximately twenty percent of familial cases are linked to various point mutations in the Cu/Zn superoxide dismutase 1 (SOD1) gene on chromosome 21 4. Transgenic mice 4,5 and rats 6 carrying mutant human SOD1 genes (G93A, G37R, G86R, G85R) have been generated, and, despite the existence of other animal models of motor neuron loss, are currently the most highly used models of the disease.
Spinal cord injury (SCI) is a heterogeneous set of conditions resulting from physical trauma to the spinal cord, with functional outcome varying according to the type, location and severity of the injury 7. Nevertheless, approximately half of human SCI cases affect cervical regions, resulting in debilitating respiratory dysfunction due to phrenic motor neuron loss and injury to descending bulbospinal respiratory axons 1. A number of animal models of SCI have been developed, with the most commonly used and clinically-relevant being the contusion 8.
Transplantation of various classes of neural precursor cells (NPCs) is a promising therapeutic strategy for treatment of traumatic CNS injuries and neurodegeneration, including ALS and SCI, because of the ability to replace lost or dysfunctional CNS cell types, provide neuroprotection, and deliver gene factors of interest 9.
Animal models of both ALS and SCI can model many clinically-relevant aspects of these diseases, including phrenic motor neuron loss and consequent respiratory compromise 10,11. In order to evaluate the efficacy of NPC-based strategies on respiratory function in these animal models of ALS and SCI, cellular interventions must be specifically directed to regions containing therapeutically relevant targets such as phrenic motor neurons. We provide a detailed protocol for multi-segmental, intraspinal transplantation of NPCs into the cervical spinal cord ventral gray matter of neurodegenerative models such as SOD1G93A mice and rats, as well as spinal cord injured rats and mice 11.

Neural precursor transplantation is a promising strategy for protecting and/or replacing lost/dysfunctional cervical phrenic motor neurons in spinal cord injury (SCI) and the motor neuron disorder, amyotrophic laterals sclerosis (ALS). We provide a protocol for cell delivery to cervical spinal cord ventral horn in rodent models of ALS and SCI.

Amyotrophic Lateral Sclerosis ve Travmatik Spinal Kord Yaralanması Servikal Ventral Horn Hedefleme intraspinal Hücre Transplantasyonu

Respiratory compromise due to phrenic motor neuron loss is a debilitating consequence of a large proportion of human traumatic spinal cord injury (SCI) ...

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis (ALS)

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 years for the sporadic disease and around 50 years for the familial disease. Due to its progressive course, 50% of the patients die within 30 months of symptom onset. In order to evaluate novel treatment options for this disease, genetic mouse models of ALS have been generated based on human familial mutations in the SOD gene, such as the SOD1 (G93A) mutation. Most important aspects that have to be evaluated in the model are overall survival, clinical course and motor function. Here, we demonstrate the clinical evaluation, show the conduction of two behavioural motor tests and provide quantitative scoring systems for all parameters. Because an in depth analysis of the ALS mouse model usually requires an immunohistochemical examination of the spinal cord, we demonstrate its preparation in detail applying the dorsal laminectomy method. Exemplary histological findings are demonstrated. The comprehensive application of the depicted examination methods in studies on the mouse model of ALS will enable the researcher to reliably test future therapeutic options which can provide a basis for later human clinical trials.

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis ALS

A mouse model for amyotrophic lateral sclerosis (ALS) is examined clinically and behaviorally. As a prerequisite for an accompanying immunohistological analysis the preparation of the spinal cord is depicted in detail.

Amyotrofik Lateral Skleroz için Fare Modeli (ALS) Klinik Test ve Omurilik Kaldırma

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 ...

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury (IRI) of the Rat Cremaster Muscle

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, intestinal ischemia as well as following transplant and cardiovascular surgery.1 Reperfusion of previously ischemic tissue, while essential for the prevention of irreversible tissue injury, elicits excessive inflammation of the affected tissue. Adjacent to the production of reactive oxygen species, activation of the complement system and increased microvascular permeability, the activation of leukocytes is one of the principle actors in the pathological cascade of inflammatory tissue damage during reperfusion.2, 3 Leukocyte activation is a multistep process consisting of rolling, firm adhesion and transmigration and is mediated by a complex interaction between adhesion molecules in response to chemoattractants such as complement factors, chemokines, or platelet-activating factor.4
While leukocyte rolling in postcapillary venules is predominantly mediated by the interaction of selectins5 with their counter ligands, firm adhesion of leukocytes to the endothelium is selectin-controlled via binding to intercellular adhesion molecules (ICAM) and vascular cellular adhesion molecules (VCAM).6, 7 
Gold standard for the in vivo observation of leukocyte-endothelial interaction is the technique of intravital microscopy, first described in 1968.8
Though various models of IRI (ischemia-reperfusion injury) have been described for various organs, 9-12 only few are suitable for direct visualization of leukocyte recruitment in the microvascular bed on a high level of image quality.8 
We here promote the digital intravital epifluorescence microscopy of the postcapillary venule in the cremasteric microcirculation of the rat 13 as a convenient method to qualitatively and quantitatively analyze leukocyte recruitment for IRI-research in striated muscle tissue and provide a detailed manual for accomplishing the technique. We further illustrate common pitfalls and provide useful tips which should enable the reader to truly appreciate, and safely perform the method.
In a step by step protocol we depict how to get started with respiration controlled anesthesia under sufficient monitoring to keep the animal firmly anesthetized for longer periods of time. We then describe the cremasteric preparation as a thin flat sheet for outstanding optical resolution and provide a protocol for leukocyte imaging in IRI that has been well established in our laboratories.

Real-time Digital Imaging of Leukocyte-endothelial Interaction in Ischemia-reperfusion Injury IRI of the Rat Cremaster Muscle

Digital intravital epifluorescence microscopy of postcapillary venules in the cremasteric microcirculation is a convenient method to gain insights into leukocyte-endothelial interaction in vivo in ischemia-reperfusion injury (IRI) of striated muscle tissue. We here provide a detailed protocol to safely perform the technique and discuss its applications and limitations.

Sıçan Cremaster&#39;ın kas iskemi-reperfüzyon Hasarı içinde Lökosit-endotelyal Etkileşim Gerçek zamanlı Dijital Görüntüleme (IRI)

Ischemia-reperfusion injury (IRI) has been implicated in a large array of pathological conditions such as cerebral stroke, myocardial infarction, ...

Tibial Nerve Transection - A Standardized Model for Denervation-induced Skeletal Muscle Atrophy in Mice

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. Although originally developed and used extensively in the rat due to its larger size, the tibial nerve in mice is big enough that it can be easily manipulated with either crush or transection, leaving the peroneal and sural nerve branches of the sciatic nerve intact and thereby preserving their target muscles. Thus, this model offers the advantages of inducing less morbidity and impediment of ambulation than the sciatic nerve transection model and also allows investigators to study the physiologic, cellular and molecular biologic mechanisms regulating the process of muscle atrophy in genetically engineered mice. The tibial nerve supplies the gastrocnemius, soleus and plantaris muscles, so its transection permits the study of denervated skeletal muscle composed of fast twitch type II fibers and/or slow twitch type I fibers. Here we demonstrate the tibial nerve transection model in the C57Black6 mouse. We assess the atrophy of the gastrocnemius muscle, as a representative muscle, at 1, 2, and 4 weeks post-denervation by measuring muscle weights and fiber type specific cross-sectional area on paraffin-embedded histologic sections immunostained for fast twitch myosin.

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of skeletal muscle atrophy. The model surgical protocol is described and demonstrated in C57Black6 mice. &#160;

Tibial Sinir transeksiyonu - Farelerde denervasyon bağlı İskelet Kas Atrofi için bir Standart Model

The tibial nerve transection model is a well-tolerated, validated, and reproducible model of denervation-induced skeletal muscle atrophy in rodents. ...

High-throughput Flow Cytometry Cell-based Assay to Detect Antibodies to N-Methyl-D-aspartate Receptor or Dopamine-2 Receptor in Human Serum

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases in children and adults. These antibodies have been used to guide diagnosis and treatment. Cell-based assays have improved the detection of antibodies in patient serum. They are based on the surface expression of brain antigens on eukaryotic cells, which are then incubated with diluted patient sera followed by fluorochrome-conjugated secondary antibodies. After washing, secondary antibody binding is then analyzed by flow cytometry. Our group has developed a high-throughput flow cytometry live cell-based assay to reliably detect antibodies against specific neurotransmitter receptors. This flow cytometry method is straight forward, quantitative, efficient, and the use of a high-throughput sampler system allows for large patient cohorts to be easily assayed in a short space of time. Additionally, this cell-based assay can be easily adapted to detect antibodies to many different antigenic targets, both from the central nervous system and periphery. Discovering additional novel antibody biomarkers will enable prompt and accurate diagnosis and improve treatment of immune-mediated disorders.

Over the recent years, live cell-based assays have been used successfully to detect antibodies against surface and conformational antigens. Here, we describe a method using high-throughput flow cytometry enabling the analysis of large cohorts of patients. Detection of novel antibodies will improve diagnosis and treatment of immune-mediated disorders.

İnsan Serum içinde N-metil-D-aspartat reseptör ya da dopamin-2 reseptör antikorları algılamak üzere yüksek verimli Flow Cytometry hücre bazlı bir deneyde

Over the recent years, antibodies against surface and conformational proteins involved in neurotransmission have been detected in autoimmune CNS diseases ...

Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are still occurring, particularly in reconstructive flap surgery. Multiple experimental flap models have been developed to analyze underlying causes and mechanisms and to investigate treatment strategies to prevent ischemic complications. The limiting factor of most models is the lacking possibility to directly and repetitively visualize microvascular architecture and hemodynamics. The goal of the protocol was to present a well-established mouse model affiliating these before mentioned lacking elements. Harder et al. have developed a model of a musculocutaneous flap with a random perfusion pattern that undergoes acute persistent ischemia and results in ~50% necrosis after 10 days if kept untreated. With the aid of intravital epi-fluorescence microscopy, this chamber model allows repetitive visualization of morphology and hemodynamics in different regions of interest over time. Associated processes such as apoptosis, inflammation, microvascular leakage and angiogenesis can be investigated and correlated to immunohistochemical and molecular protein assays. To date, the model has proven feasibility and reproducibility in several published experimental studies investigating the effect of pre-, peri- and postconditioning of ischemically challenged tissue.

The window of the murine dorsal skinfold chamber presented visualizes a zone of acute persistent ischemia of a musculocutaneous flap. Intravital epi-fluorescence microscopy permits for direct and repetitive assessment of the microvasculature and quantification of hemodynamics. Morphologic and hemodynamic results can further be correlated with histological and molecular analyses.

Mouse Dorsal Skinfold Odası&#39;nda İskemik Doku Yaralanması: Bir Cilt Flap Model Akut Kalıcı iskemisi Araştırma

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are ...

Single-stage Dynamic Reanimation of the Smile in Irreversible Facial Paralysis by Free Functional Muscle Transfer

Unilateral facial paralysis is a common disease that is associated with significant functional, aesthetic and psychological issues. Though idiopathic facial paralysis (Bell&#8217;s palsy) is the most common diagnosis, patients can also present with a history of physical trauma, infectious disease, tumor, or iatrogenic facial paralysis. Early repair within one year of injury can be achieved by direct nerve repair, cross-face nerve grafting or regional nerve transfer. It is due to muscle atrophy that in long lasting facial paralysis complex reconstructive methods have to be applied. Instead of one single procedure, different surgical approaches have to be considered to alleviate the various components of the paralysis.
The reconstruction of a spontaneous dynamic smile with a symmetric resting tone is a crucial factor to overcome the functional deficits and the social handicap that are associated with facial paralysis. Although numerous surgical techniques have been described, a two-stage approach with an initial cross-facial nerve grafting followed by a free functional muscle transfer is most frequently applied. In selected patients however, a single-stage reconstruction using the motor nerve to the masseter as donor nerve is superior to a two-stage repair. The gracilis muscle is most commonly used for reconstruction, as it presents with a constant anatomy, a simple dissection and minimal donor site morbidity.
Here we demonstrate the pre-operative work-up, the post-operative management, and precisely describe the surgical procedure of single-stage microsurgical reconstruction of the smile by free functional gracilis muscle transfer in a step by step protocol. We further illustrate common pitfalls and provide useful tips which should enable the reader to truly comprehend the procedure. We further discuss indications and limitations of the technique and demonstrate representative results.

The use of the masseteric nerve as donor nerve represents a single-stage alternative to the criterion standard two-stage procedure of cross-facial nerve grafting and free muscle transfer in facial paralysis. We provide a detailed description to safely perform this technique with a gracilis muscle transfer and discuss indications and limitations.

Ücretsiz Fonksiyonel kas transferi ile geri dönüşümsüz Yüz felci içinde Smile Tek aşamalı Dinamik Reanimasyon

Unilateral facial paralysis is a common disease that is associated with significant functional, aesthetic and psychological issues. Though idiopathic ...

A Method of Trigonometric Modelling of Seasonal Variation Demonstrated with Multiple Sclerosis Relapse Data

This report describes a novel Stata-based application of trigonometric regression modelling to 55 years of multiple sclerosis relapse data from 46 clinical centers across 20 countries located in both hemispheres. Central to the success of this method was the strategic use of plot analysis to guide and corroborate the statistical regression modelling. Initial plot analysis was necessary for establishing realistic hypotheses regarding the presence and structural form of seasonal and latitudinal influences on relapse probability and then testing the performance of the resultant models. Trigonometric regression was then necessary to quantify these relationships, adjust for important confounders and provide a measure of certainty as to how plausible these associations were. Synchronization of graphing techniques with regression modelling permitted a systematic refinement of models until best-fit convergence was achieved, enabling novel inferences to be made regarding the independent influence of both season and latitude in predicting relapse onset timing in MS. These methods have the potential for application across other complex disease and epidemiological phenomena suspected or known to vary systematically with season and/or geographic location.

Combining plot analysis with trigonometric regression is a robust method for exploring complex, cyclical phenomena such as relapse onset timing in multiple sclerosis (MS). This method enabled unbiased characterisation of seasonal trends in relapse onset permitting novel inferences around the influence of seasonal variation, ultraviolet radiation (UVR) and latitude.

Multipl Skleroz Relaps Verilerle kanıtlanmış Mevsimsel Değişim Trigonometrik Modelleme Bir Yöntem

This report describes a novel Stata-based application of trigonometric regression modelling to  55 years of multiple sclerosis relapse data from 46 ...

Design, Fabrication, and Administration of the Hand Active Sensation Test (HASTe)

The concept of personalizing neurologic rehabilitation, based on individual impairments, has experienced a recent surge. In parallel, the number of outcome measures of upper extremity motor performance has grown. However, clinicians and researchers lack practical, quantitative measures of the hand&#8217;s natural role as a receptor of the environment. The Hand Active Sensation Test (HASTe), developed by Williams and colleagues in 2006, is a valid and reliable measure of haptic performance. Though not available commercially, the HASTe can be fabricated from inexpensive materials, and it has been used successfully to identify impairments in haptic touch in individuals with stroke. (Williams, 2006). This paper presents the methods of design and fabrication of the HASTe testing kit, as well as a visual screen to be used during administration, and instructions for the tests administration and scoring.

Design, Fabrication, and Administration of the Hand Active Sensation Test HASTe

The Hand Active Sensation Test (HASTe) is a valid and reliable measure of haptic performance, which has been used successfully to identify impaired haptic touch in individuals with stroke. The purpose of this paper is to describe the design, fabrication and administration of the HASTe.

Tasarım, ve Yönetim El Fabrikasyon Aktif Sensation Testi (Haste)

The concept of personalizing neurologic rehabilitation, based on individual impairments, has experienced a recent surge. In parallel, the number of ...