Financiación apoyo del Instituto de Investigación de Enfermedades de la Neurona Motora de Australia (MNDRIA), Sylvia y Charles Viertel Charitable Foundation subvención Investigador Clínico, Fundación Ramaciotti y Salud Nacional y el Consejo de Investigación Médica de Australia (número de concesión del proyecto APP1024915) Se agradece.

1. Preparación del paciente <ol><li> Reclutar a los pacientes de forma prospectiva y consecutiva. Nota: El siguiente protocolo es aprobado por los Comités de Ética de Sydney West Area de Servicios de Salud Humanos de Investigación. Obtuvo el consentimiento informado de todos los pacientes. Los pacientes fueron reclutados prospectiva y consecutiva de la clínica ELA / neuromuscular en el Hospital de Westmead. </li><li> Determinar la idoneidad de los pacientes con ELA para la prueba. Excluir los pacientes que no son diagnosticados con ELA o un trastorno neuromuscular. </li><li> Excluir los pacientes con una neuropatía focal coexistente, neuropatía mediano en la muñeca o neuropatía cubital en el codo. </li><li> Excluir los pacientes con una neuropatía generalizada, como la polineuropatía diabética. </li><li> Ampliamente investigar y clínicamente los pacientes de seguimiento para confirmar el diagnóstico de ELA o trastornos neuromusculares imitan. </li><li> Asegúrese de que el consentimiento informado es proporcionada por todos los pacientes, incluyendo la ELA y los pacientes con trastorno imitan neuromusculares, de all los procedimientos neurofisiológicos. </li></ol> 2. Evaluación clínica <ol><li> Determine la clasificación clínica de todos los pacientes con ELA mediante la Esclerosis Lateral Amiotrófica Functional Rating Scale-Revised (ALSFRS-R) anotar 18. </li><li> Determinar la tasa de progresión de la enfermedad en todos los pacientes de ALS de acuerdo a la fórmula se informó anteriormente; (48-ALSFRS-R) / duración de los síntomas) 19. </li><li> Determine la fuerza muscular en los pacientes que utilizan la escala del Medical Research Council (MRC) Evaluación 20. Desarrollar una puntuación total MRC que comprende el miembro superior (hombro secuestro; flexión del codo; extensión del codo, flexión dorsal de la muñeca; abducción de los dedos; abducción del pulgar, la puntuación total de 60) y el miembro inferior (flexión de la cadera, flexión dorsal del tobillo, la puntuación total de 30, de extensión de rodilla) grupos musculares . La puntuación total MRC debe ser 90 si la fuerza muscular es normal. </li><li> Determinar el sitio de inicio de la enfermedad, definida como bulbar o extremidad, en todos los ALSpacientes. </li></ol> 3. Evaluación neurofisiológica <ol><li> Llevar a cabo estudios de conducción del nervio motor de los nervios mediano y cubital con un potencial de acción muscular compuesto (CMAP) las respuestas registradas en el abductor corto del pulgar (APB), los músculos interóseos primera dorsal (IED) y la minimi dígitos secuestrador (ADM). </li><li> Preparar los sitios de estimulación en la muñeca, por la limpieza de la superficie de la piel con un gel abrasivo para reducir la resistencia de la piel, seguido de la aplicación de un paño con alcohol. Ajuste la corriente de estimulación a un 20% por encima de la intensidad requerida para producir una respuesta CMAP máxima (corriente supramáxima). </li><li> Asegúrese de que el sitio de estimulación seleccionado exhibe el umbral más bajo para la estimulación de tal manera que se evita el volumen de conducción. </li><li> Preparar el sitio de grabación sobre cada músculo por la limpieza de la superficie de la piel con un gel abrasivo para reducir la resistencia de la piel, seguido de la aplicación de un paño con alcohol. </li><li> Anote el tamaño de la acti motor compuestoen el potencial (CMAP) de los músculos de la APB, ADM, y la IED. Las respuestas que se deben registrar en un 10 mm electrodos de disco de oro colocados en un arreglo tendón vientre sobre cada músculo. Específicamente, posicionar el electrodo activo sobre el punto medio del músculo respectiva asegurar un despegue negativo de la respuesta CMAP, mientras que el electrodo de referencia que se coloca sobre la base del pulgar (APB y grabaciones IED CMAP) y la base de dígitos 5 (para ADM CMAP grabaciones). </li><li> Ajuste la distancia entre el cátodo y el electrodo estimulante activo para APB y ADM de acción motor compuesto posibles respuestas a 5 cm y la distancia a la inversión extranjera directa a los 8 cm. </li><li> Coloque la placa de tierra del neutro de electrocirugía en la cara dorsal de la mano, entre el estímulo y G1 electrodo con gel conductor para reducir el artefacto. Antes de la aplicación de la tierra neutra preparar el sitio por la limpieza de la superficie de la piel con un gel abrasivo para reducir la resistencia de la piel, seguido por la aplicación de un paño con alcohol. </li><li>Asegúrese de que los ajustes de filtro son entre 3 Hz (filtro de baja frecuencia) y 10 KHz (filtro de alta frecuencia). </li><li> Asegúrese de que la velocidad de barrido se ajusta a 20 ms o 2 ms / división. </li><li> Asegúrese de que la sensibilidad de las respuestas CMAP grabación está ajustado a 5 mV. </li><li> Monitor de temperatura en el sitio de la estimulación durante todo el estudio y asegurarse de que la temperatura del miembro se mantiene a 32 ° C. </li></ol> 4. Análisis e Interpretación <ol><li> Medir amplitudes-línea de base a pico CMAP (mV) de los músculos del APB, la IED y las ADM en pacientes con ELA y los controles de la enfermedad imitan neuromusculares. </li><li> Calcular el índice de división mano (Figura 1) multiplicando la amplitud de CMAP registrado en los músculos APB y de IED, y dividiendo este producto por la amplitud de CMAP registrada sobre el músculo ADM, de la siguiente manera: SI = APB CMAP * IED CMAP _________________________ &amp; #160; ADM CMAP </li><li> Determinar la utilidad diagnóstica del índice de mano dividida mediante la comparación de los valores de la IS entre los pacientes con ELA y controles patológicos trastorno imitan neuromusculares de acuerdo con las normas para la presentación de informes de diagnóstico de la enfermedad (STARD) criterios. </li><li> Determinar los valores de corte de diagnóstico óptimas para el índice de la mano dividida utilizando el funcionamiento del receptor (ROC) curvas características. Derivar curva ROC por el trazado de la sensibilidad (eje y) y 1-una especificidad (eje x) para valores de SI derivados de ALS y pacientes trastorno neuromuscular. </li></ol>

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Los autores no tienen nada que revelar.

El presente estudio informa sobre la utilidad diagnóstica del índice de fractura mano en la ELA, un nuevo biomarcador de diagnóstico neurofisiológico. El SI distinguió fiablemente ALS de trastornos neuromusculares, con un valor de corte diagnóstico óptimo de 5,2. La reducción en el SI fue más prominente en las extremidades de aparición pacientes con ELA. Es importante destacar que una proporción sustancial de los pacientes con ELA que se clasificaron en la categoría de diagnóstico &quot;posible&quot; de acu...

La mano dividida, se refiere a la atrofia preferencial del complejo grupo tenar de los músculos intrínsecos de la mano [abductor corto del pulgar (APB) y primer interóseo dorsal (IED)], con relativa preservación de los músculos hipotenar, y es una característica clínica específica y precoz de amiotrófica esclerosis lateral (ALS) 1-4. El hallazgo de que el signo de división mano es específica para la ELA, sugiere un posible papel para el signo de división mano como un biomarcador de diagnóstico en la ELA 3. La cuantificación de la señal dividida lado, a través del desarrollo de un nuevo biomarcador neurofisiológico, puede reducir aún más la ayuda en el diagnóstico de la ELA. Específicamente, el índice de división mano (SI), que cuantifica el fenómeno de división lado, se obtiene multiplicando el potencial de acción muscular compuesto (CMAP) de amplitud registrada en los músculos tenares complejos (APB y IED), y dividiendo este producto por la amplitud de CMAP registrado durante el hypmúsculos othenar (a saber, el abductor del meñique, ADM) 5. El diagnóstico de la ELA se basa en gran parte en criterios basados ​​clínicamente que abarcan una combinación de la neurona motora superior e inferior firma 6. Estos criterios, sin embargo, se consideraron insensible especialmente en el establecimiento de un diagnóstico de ALS en las primeras etapas de la 7-10 proceso de la enfermedad. Una modificación reciente de los criterios de diagnóstico fueron desarrolladas 11, y aunque estos criterios parecen aumentar la sensibilidad de diagnóstico 12-16, el aumento de la sensibilidad parece restringida a bulbar de inicio 15 pacientes con ELA. En ausencia de una prueba patognomónica, el diagnóstico de la ELA se puede retrasar significativamente 8. En última instancia, la institución de las terapias neuroprotectoras y el reclutamiento en ensayos clínicos puede ser retrasado, tal vez más allá del período de ventana terapéutica crítica 9, 17. En consecuencia, lautilidad de diagnóstico de la IS se evaluó de forma prospectiva en pacientes con ELA esporádica.

<table border="0" cellpadding="0" cellspacing="0">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:125px;"></td>
			<td style="width:120px;"></td>
			<td style="width:136px;"></td>
			<td style="width:203px;"></td>
		</tr>
		<tr height="20">
			<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>
			<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;">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>
			<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;">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>
			<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;">Amyotrophic lateral sclerosis rating scale-revised questionnaire</td>
			<td>Publication&#160;</td>
			<td>Experimental builder</td>
			<td>Stage disease</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td colspan="2">(see Methods and References)</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;">Medical Research Council muscle strength scale</td>
			<td>Medical Research Council-UK</td>
			<td>Experimental builder</td>
			<td>Stage disease</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;"></td>
			<td colspan="2">(see Methods and References)</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;"></td>
			<td></td>
			<td></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 colspan="2" height="20" style="height:20px;">Table of equipment used</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Name of the equipment used</td>
			<td>Company</td>
			<td>Catalog number</td>
			<td>Comments</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;">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

El fenómeno de división mano se refiere a la emaciación predominante de la eminencia tenar músculos y es una característica temprana y específica de la esclerosis lateral amiotrófica (ALS). Un índice novedoso mano dividida (SI) fue desarrollado para cuantificar el fenómeno de división lado, y su utilidad diagnóstica fue evaluada en pacientes con ELA. El índice de división mano se obtiene dividiendo el producto del potencial de acción muscular compuesto (CMAP) de amplitud registrado durante el abductor corto del pulgar y la primera dorsal músculos interóseos por la amplitud de CMAP grabada sobre el músculo abductor del meñique. Con el fin de evaluar la utilidad diagnóstica del índice de fractura lado, los pacientes con ELA fueron evaluados prospectivamente y sus resultados se compararon con los pacientes con trastorno neuromuscular. El índice de división mano se redujo significativamente en la ELA, en comparación con los pacientes con trastorno neuromuscular (P &lt;0,0001). Miembro de aparición pacientes con ELA mostraron la mayor reducción en el índice de división mano, y un valorde 5,2 o menos diferenciadas fiable ALS de otros trastornos neuromusculares. En consecuencia, el índice de división mano parece ser un nuevo biomarcador de diagnóstico para la ELA, quizás facilitando un diagnóstico más temprano.

Fenotipo clínico En total, se estudiaron 44 pacientes con ELA, de los cuales el 76% (N = 33) fueron clasificados como definitiva o probable, y el 24% (N = 11) como posibles ALS acuerdo con los criterios de Awaji 11. El diagnóstico de ELA se confirmó en la cohorte &quot;posible&quot; después de extensas investigaciones y el seguimiento clínico durante un máximo de 3 años y 53% murieron durante este período. Enfermedad bulbar inicio fue evidente en el 41%, mientras qu...

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

La atrofia de los músculos intrínsecos disociada mano, denominada mano dividida, parece ser una característica específica de la esclerosis lateral amiotrófica (ALS). En consecuencia, un nuevo test diagnóstico neurológico, denominado el índice de división lado, fue desarrollado para cuantificar el fenómeno clínico de la mano dividida, que diferenciaba ALS trastornos imitan.

Utilidad de la disociada intrínseca Mano Atrofia muscular en el diagnóstico de la esclerosis lateral amiotrófica

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 vistas.  Westmead Hospital.  La atrofia de los músculos intrínsecos disociada mano, denominada mano dividida, parece ser una característica específica de la esclerosis lateral amiotrófica (ALS). En consecuencia, un nuevo test diagnóstico neurológico, denominado el índice de división lado, fue desarrollado para cuantificar el fenómeno clínico de la mano dividida, que diferenciaba ALS trastornos imitan. 

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.

Un Protocolo de Evaluación Integral de la disfunción bulbar de la Esclerosis Lateral Amiotrófica (ALS)

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

Investigación

Medicina

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.

Intraespinal trasplante de células de Orientación asta ventral del cuello uterino en la esclerosis lateral amiotrófica y la lesión de la médula espinal

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.

Los ensayos clínicos y de eliminación de la médula espinal en un modelo murino de esclerosis lateral amiotrófica (ELA)

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.

En tiempo real de imágenes digitales de los leucocitos del endotelio interacción en el daño por isquemia-reperfusión (IRI) del músculo cremáster Rata

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;

Transección del nervio tibial - Un modelo normalizado para la atrofia del músculo esquelético denervación inducida en ratones

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.

Ensayo de alto rendimiento basado en células citometría de flujo para detectar anticuerpos a la N-metil-D-aspartato receptor o dopamina-2 del receptor en suero humano

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.

Lesiones tejido isquémico en la Cámara de los pliegues cutáneos dorsal del ratón: Un modelo de la piel de la aleta para Investigar la isquemia aguda persistente

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.

De una etapa dinámica Reanimación de la Sonrisa en Irreversible Parálisis facial por Free transferencia muscular funcional

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.

Un método de trigonométricas Modelización de Variación Estacional demostrada con Esclerosis Múltiple La recaída de datos

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.

Diseño, Fabricación y Administración de la Mano Activo Prueba Sensation (Haste)

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