This work was supported by the National Institutes of Health (HL114669). CM is a recipient of a postdoctoral research fellowship grant awarded by the Myocarditis Foundation, NJ.

ÉTICA DE MOTIVOS: Todos los procedimientos con animales se realizaron de conformidad con las directrices para el Cuidado y Uso de Animales de Laboratorio y aprobados por la Universidad de Nebraska-Lincoln, Lincoln, NE. 1. La inducción de miocarditis autoinmune experimental <ol><li> Preparar la solución de péptido disolviendo MyHC-α 334-352 en 1x solución salina tamponada con fosfato a una concentración final de 2 ml mg/1.5. </li><li> Preparar la toxina de pertussis (PT) mediante la adición de 1 ml de 1x PBS estéril a un vial que contiene 50 g de PT se liofilizó para obtener una concentración de solución madre de 50 ng / l. Tomar 20 l de la población en un tubo estéril de 1,5 ml y añadir 980 l de 1x PBS estéril para obtener una concentración de trabajo de 1 ng / l. </li><li> Preparar adyuvante completo de Freund (CFA) mediante la adición de 40 mg de Mycobacterium tuberculosis H37Rv (MTB) extraer a 10 ml de CFA para obtener una concentración finalde 5 mg / ml. </li><li> Preparar la emulsión péptido CFA. NOTA: Para la inducción de EAM, la emulsión péptido CFA se administró en 150 l que contenía 100 g de MyHC-α 334-352 por animal. Por ejemplo, para inmunizar a diez ratones utilizan 1,5 ml de emulsión de péptido-CFA que contiene 1 mg de MyHC-α 334-352. <ol><li> Para preparar 1,5 ml de emulsión, parte alícuota de 750 l de MyHC-α solución de péptido 334 a 352 en un tubo de 1,5 ml, y CFA complementan con MTB en otro tubo de 1,5 ml. Usando una jeringa de 3 ml luer-lok, señalar a la solución de péptido primero, seguido por el extracto de CFA / MTB. </li><li> Conecte la jeringa a una llave de paso de 3 vías y conectar la otra salida de la llave de paso a un vacío ml jeringa 3. Ajuste de la permeabilidad de la llave de paso de modo que la mezcla de péptido-CFA fluye de una jeringa a la otra con razonablemente buena resistencia. </li><li> Mezclar empujando el contenido de una jeringa a la otra repetidamente para ~ 1 min y luego se coloca todo el conjunto en hielo durante ~ 3 min. Repita esteprocedimiento de un mínimo de 10 veces. </li><li> Determinar la consistencia de la emulsión mediante la colocación cuidadosamente una pequeña gota en el agua inmóvil en un vaso de precipitados de vidrio de 100 ml. No se espera que la gotita de dispersar en agua. Si lo hace, continuar mezclando hasta que se consigue la consistencia deseada. </li><li> Transferir el contenido de emulsión de 3 ml en jeringas de 1 ml jeringas Luer Lok mediante la sustitución de una de las dos jeringas de 3 ml unidos a la llave de paso con la jeringa de 1 ml, y adjuntar una aguja 27 ½ G para la jeringa de 1 ml. </li></ol></li><li> Inyectar 150 ml de la emulsión péptido CFA en dosis divididas por vía subcutánea en ambas regiones inguinales de seis a ocho semanas de edad las mujeres ratones A / J (~ 75 l cada uno). </li><li> Administrar 100 l de suspensión de PT (100 ng) por vía intraperitoneal a cada animal en el día 0 y el día 2 después de la vacunación. </li><li> Repita el procedimiento de inmunización en el día 7 por la administración de 150 l de péptido-CFA emulsión en dosis divididas por vía subcutánea en ambos lados de tél esternón (~ 75 l cada uno). Preparar esta emulsión fresca como anteriormente. Luego, el día 21, someter a los animales a imágenes MRM, consulte el paso 3. </li></ol> 2. Manejo Animal <ol><li> Coloque cada ratón en una cámara de inducción anestésica que contiene 2% de mezcla de isoflurano al aire con una almohadilla térmica colocada debajo para mantener el calor, y transferir el animal a un titular de animales especialmente diseñado (Figura 1). </li><li> Inmovilizar el animal en la posición boca abajo en el soporte de animal para su hocico encaja en el cono de la nariz para mantener la anestesia (Figura 1). Fije la cabeza del ratón con una barra mordida unidos a los dientes delanteros del ratón. </li><li> Encienda el calentador de soplado de aire con su manguera de salida se inserta en el taladro vertical del escáner para mantener la temperatura corporal del animal durante el experimento. </li><li> Mantener la anestesia en 0,5 a 2% de isoflurano con una velocidad de flujo de 2 ml / min durante toda la sesión de formación de imágenes. Confirmaranestesia utilizando el método pizca dedo del pie, no hay movimiento esperado. </li><li> Establecer un sensor neumático almohada, el ratón de cola / tobillo sensor de oximetría de pulso de fibra óptica, y la sonda de temperatura rectal para monitorear la respiración, la frecuencia cardiaca y la temperatura corporal, respectivamente (Figura 1). Nota: gating cardíaco se realiza a través de la oximetría de pulso, que permite la monitorización no invasiva de la saturación arterial de oxígeno en sangre. El sensor de oximetría de pulso debe estar unido al tobillo izquierdo, y el pie debe ser asegurado con un bucle de hilo y cinta adhesiva para mantener el tobillo en el sensor. De formación de imágenes de MRM se logra por gating la señales cardiacas respiración y sin el uso de cualquier agente de contraste. </li></ol> 3. Adquisición de imágenes <ol><li> Después de preparar al animal (Figura 1), colocar el ratón en el centro del escáner de MRM con el corazón posicionado en el centro del campo de visión (FOV), donde el campo magnético es máxima homogeneidad. NOTA: Muchas de ánima(89 mm) 9,4 T imán vertical taladro equipado con gradientes ejes triples de 100 G / cm y 4 cm de radio-frecuencia (RF) de la bobina de imagen se va a utilizar para obtener imágenes de alta resolución en tres dimensiones (3D). Nota: Asegúrese de que los suministros no magnéticos cuando se utiliza un escáner de resonancia magnética. </li><li> Ejecutar la interfaz de imagen y seleccionar &quot;Nuevo Estudio&quot; en el menú &quot;Opciones de estudio&quot;. Escriba &quot;mtune&quot; en la barra de comandos y ejecutarlo para levantar el &quot;Tune GUI&quot;. A continuación, seleccione &quot;Start Sonda Tune&quot; y haga clic en el botón &quot;Escala automática&quot;. La GUI Tune cambiará para mostrar la RFsignal. Utilice los mandos de sintonía / partido remotas en el extremo de la bobina para sintonizar la bobina de RF a la frecuencia de resonancia de protones (400 MHz). En la pestaña &quot;Inicio&quot;, vaya a la página &quot;Prescan&quot; para ejecutar la calibración de frecuencia y potencia haciendo clic en los botones correspondientes. Pulsa el botón de XYZ (rápida) en la &quot;ficha&quot; de &quot;Shim Estudio pestaña para tirar de la página calce.Ir a la página de cuña, seleccionar todas las iteraciones, y pulsa el botón para realizar shim calce automático. </li><li> Seleccione una secuencia de explorador desde la pestaña &quot;Estudio&quot; de la interfaz de imágenes para localizar el corazón de ratón. En la pestaña &quot;Acquire&quot; cambiar el campo de visión de 35 mm 2 y mantener la configuración predeterminada de la máquina. Haga clic en Inicio para ejecutar la secuencia; ajustar la posición del soporte de animal si el corazón no está en el centro de campo de visión FOV, volver a sintonizar la bobina de RF y obtener las imágenes de exploradores de nuevo. </li><li> Haga clic en el botón de secuencia de &quot;joyas&quot; en la pestaña de &quot;Estudio&quot; y luego introducir los parámetros de adquisición en la pestaña &quot;Acquire&quot; correspondiente a la obtención de dos planos ortogonales a lo largo del eje corto y eje largo del corazón 20,21. NOTA: los parámetros de adquisición típicas para una exploración eco de gradiente son: grosor de corte, 1 mm; tiempo de repetición (TR), 200 ms; tiempo de eco (TE), 2,67 ms (mínimo); ángulo de inclinación, 25 °; tamaño de la matriz de rigidización, 128 x 128; FOV, 22 mm2; número de adquisiciones, 4; y un tiempo de adquisición aproximada de 1 min y 30 seg. </li><li> En la pestaña &quot;Avanzado&quot;, seleccione la secuencia &quot;CINE&quot; para recoger los pulsos de eje corto de cine de las imágenes de RM-oximetría cerrada con el fin de medir los parámetros anatómicos y funcionales del VI. Ajuste la posición y el ángulo de los cortes de imagen basado en el eje largo del corazón solo pasar el mouse y arrastrando. Introduzca los siguientes parámetros de imagen en la ficha &quot;Acquire&quot; para obtener la secuencia de eco de gradiente de cine: TR, 500 ms; TE, 5 mseg; FOV, 22 mm 2; matriz de adquisición, 256 x 256; grosor de corte, 1 mm; número de adquisiciones, 8; número de tramas, 6; y un tiempo de adquisición de ~ 17 min. Haga clic en Iniciar para iniciar la adquisición. </li><li> Convertir las imágenes adquiridas a formato DICOM utilizando la pestaña &quot;I / O&quot; del software de imagen y transferir los archivos correspondientes a los centros de datos para su procesamiento. </li><li> Al final del procedimiento de imagen, todoOW los ratones se recuperen de la anestesia dentro de las jaulas de filtro-top. No deje que los ratones sin vigilancia hasta que recobren el conocimiento suficiente para mantener decúbito esternal y mantenerlos para otros estudios, según sea necesario. </li></ol> 4. Análisis de datos de resonancia magnética cardiaca de imágenes de microscopía <ol><li> Utilice software de Segmento para analizar los parámetros anatómicos y funcionales del VI 19. Al cargar el formato DICOM las imágenes de cine en el software utilizando el &quot;Archivo de imagen abierta (s)&quot; submenú del menú &quot;Archivo&quot;. En la GUI seleccione &#39;MRGE &quot;como la técnica de imagen,&#39; Cine &#39;como el tipo de imagen y&#39; de eje corto medioventricular&quot; como el plano de la vista de la imagen. </li><li> Especifique los marcos de tiempo que se utilizará para la telediástole y al final de la sístole a través del &quot;Set Plazo actual para poner fin a Diástole&quot; y &quot;Set Plazo actual para poner fin a la sístole&quot; submenús del menú &quot;Editar&quot;, respectivamente. </li><li>Primero haga clic en el botón de comando &quot;LV&quot; y luego el botón de comando &quot;PAI&quot; en el panel inferior derecho &quot;ENDO&quot; y para dibujar manualmente el endocardio y el epicardio del ventrículo izquierdo, respectivamente. Quitar los músculos papilares pulsando el botón de comando correspondiente con el fin de aumentar la precisión de los cálculos. </li><li> Leer los parámetros LV cuantificados, como el volumen diastólico, volumen sistólico, el volumen sistólico y la fracción de eyección en el panel superior derecho. Haga clic en el botón de comando &quot;Misc&quot; y luego el botón de comando &quot;medición del calibre&quot; para medir los parámetros de baja tensión tales como el grosor de la pared, y el diámetro ventricular 22. </li><li> Haga clic en la opción &quot;Guardar imagen Tanto Pilas y Segmentación&quot; submenú del menú &quot;Archivo&quot; para guardar las imágenes, incluidas las segmentaciones, en &#39;. Mat&#39; Formato de volver a procesar las imágenes según sea necesario. </li></ol>

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No conflicts of interests were declared.

Este estudio describe el procedimiento de MRM y su utilidad como una herramienta no invasiva para determinar anomalías cardíacas en ratones afectados con miocarditis autoinmune. Dado que las características histológicas de EAM se asemejan a la miocarditis postinfeccioso de los seres humanos, los modelos de ratón se emplean comúnmente para delinear los mecanismos inmunes de las lesiones del miocardio 23-25. Sin embargo, los animales afectados con miocarditis parecen clínicamente normales, y el diagnóst...

La insuficiencia cardíaca es la principal causa de las muertes, y miocarditis es una causa predominante de la insuficiencia cardíaca en los jóvenes adolescentes 1. La mayoría de los pacientes afectos de miocarditis permanecen asintomáticos y la enfermedad se resuelve espontáneamente 2. Sin embargo, el 10-20% de las personas afectadas pueden desarrollar enfermedades crónicas, lo que lleva a la miocardiopatía dilatada (DCM) 3. Se han desarrollado diversos modelos animales para estudiar la patogénesis inmune de la miocarditis. La enfermedad puede ser inducida en-miocarditis susceptibles de A / J y ratones Balb / c mediante la inmunización de los animales con la miosina cardíaca de la cadena pesada (MyHC)-α o sus fragmentos de péptidos inmunodominantes o mediante la infección con patógenos como el virus Coxsackie B3 4-9. Este estudio implica MyHC-α miocarditis inducida-334-352 en ratones A / J. A pesar de mostrar las infiltraciones miocárdicas, los animales miocarditis afectadas parecen clínicamente normal; diagnóstico se basa en la evaluación histológica de corazones para la inflamación 7 unND ecocardiografía 10. Microscopía de resonancia magnética (MRM) es un método comúnmente utilizado para obtener imágenes cardiovasculares con aviones de alta resolución en tres dimensiones, que permita la verificación de los detalles funcionales a nivel de los vasos sanguíneos minutos (hasta 10 m de diámetro), pero este nivel de poder de resolución es no alcanzable con la formación de imágenes de resonancia magnética de rutina (IRM) procedimiento de exploración, en el que, la resolución se obtiene generalmente de hasta 1 mm 11-14. MRM ofrece una ventaja, ya que permite la adquisición de imágenes de alta resolución y también para derivar los parámetros de rendimiento en los puntos de tiempo de proceso de la enfermedad 14. Clínicamente, las imágenes MRM se ha aplicado ampliamente para estudiar los parámetros funcionales del corazón enfermo, pulmón o el cerebro 15-17. En este estudio, se muestra el uso de una técnica de MRM como una herramienta no invasiva para determinar anomalías cardíacas en ratones A / J afectadas con miocarditis autoinmune. Específicamente, tél imágenes MRM permite la cuantificación de los parámetros funcionales como ventrículo izquierdo (LV) el volumen diastólico final y la fracción de eyección (FE) con una precisión razonable 18. Las definiciones de los parámetros respectivos son: LV volumen diastólico final, el volumen de sangre en el ventrículo izquierdo al final del ciclo diastólico, y la fracción de eyección, el volumen sistólico / volumen diastólico final. El análisis de los datos se realiza mediante el software desarrollado por segmentos de libre disposición para procesamiento de imágenes cardiovasculares compatible con DICOM adquiridos por los escáneres de resonancia magnética 19. Los datos revelaron un aumento en el grosor de la pared del VI en los animales miocárdico, que corresponde a una disminución en el volumen diastólico final del LV, el volumen sistólico y la fracción de eyección, en comparación con estos parámetros funcionales en ratones sanos.

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			<td style="width:364px;">Neopeptide, Cambridge, MA</td>
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			<td style="width:167px;">Store at 4&#959;C</td>
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			<td style="width:364px;">Sigma Aldrich, St Louis, MO</td>
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			<td>Store at 4&#959;C</td>
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			<td>231141</td>
			<td>Store at 4&#959;C</td>
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			<td>List Biologicals Laboratories, Campbell, CA</td>
			<td>181</td>
			<td>Store at 4&#959;C</td>
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			<td>Corning, Manassas, VA</td>
			<td>21-040-CV</td>
			<td>Store at 4&#959;C</td>
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			<td>Piramal Healthcare, Mumbai, India</td>
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			<td height="21" style="height:21px;">Leur-lok sterile 3 ml syrringe</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>309657</td>
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			<td height="21" style="height:21px;">Sterile needle, 18 G</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>305195</td>
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			<td height="21" style="height:21px;">Sterile needle, 27 1/2 G</td>
			<td>BD, Franklin Lakes, NJ</td>
			<td>305109</td>
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			<td>Smiths Medical ASD, Inc. Dublin, OH</td>
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			<td height="21" style="height:21px;">Kerlix gauze bandage rolls&#160;</td>
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			<td>Theramics Corporation, Springfield, IL</td>
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			<td height="21" style="height:21px;">Reptile heating lamp&#160;</td>
			<td>Energy Savers Unlimited, Inc. Carson, CA</td>
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			<td height="21" style="height:21px;">Cotton tipped applicator, 6&#8217;&#8217; wooden stem&#160;</td>
			<td>Jorgensen Laboratories, Inc. Loveland, CO</td>
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			<td height="21" style="height:21px;">Summit Anesthesia Support system for regulating flow of anesthesia&#160;</td>
			<td>Summit Anesthesia Solutions, Ann Harbor, MI</td>
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			<td>Agilent Technologies, Santa Clara, CA</td>
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			<td height="21" style="height:21px;">Signal breaking module, MR compatible small animal monitoring and gating system</td>
			<td>Small Animal Instruments, Inc. Stony Brook, NY</td>
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			<td>Small Animal Instruments, Inc. Stony Brook, NY</td>
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			<td height="21" style="height:21px;">Quantitative Medical Image analysis software&#160;</td>
			<td colspan="3">http://segment.heiberg.se;&#160; Segment v1.8 R1430,&#160; Medviso, Oresunds region, Sweden</td>
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			<td height="21" style="height:21px;">Matlab software&#160;</td>
			<td>The Mathworks, Inc.&#160; Natick, MA</td>
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			<td height="21" style="height:21px;">Computer-Unix operating system</td>
			<td style="width:364px;"></td>
			<td style="width:145px;"></td>
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noninvasive assessment of cardiac abnormalities in experimental autoimmune myocarditis by magnetic resonance microscopy imaging in the mouse

La miocarditis es una inflamación del miocardio, pero sólo ~ 10% de los afectados muestran manifestaciones clínicas de la enfermedad. Para estudiar los eventos inmunes de lesiones de miocardio, diversos modelos de ratón de la miocarditis se han utilizado ampliamente. Este estudio implicó la miocarditis autoinmune experimental (EAM) inducida con miosina cardíaca de la cadena pesada (MyHC)-α 334-352 en ratones A / J; los animales afectados desarrollan miocarditis linfocítica pero sin signos clínicos aparentes. En este modelo, la utilidad de la microscopía de resonancia magnética (MRM) como una modalidad no invasiva para determinar los cardíacas cambios estructurales y funcionales en los animales inmunizados con MyHC-α 334-352 se muestra. EAM y ratones sanos fueron imágenes utilizando un 9.4 T (400 MHz) 89 mm escáner orificio central vertical equipado con una sonda de imagen radiofrecuencia 4 cm milpiés y 100 g cm gradientes ejes / triples. Imágenes cardíacas fueron compradas a los animales anestesiados utilizando una secuencia de pulsos de cine basado gradiente-eco, y el ánimaLS fueron controlados por la respiración y la oximetría de pulso. El análisis reveló un aumento en el espesor de la pared ventricular en ratones EAM, con la correspondiente disminución en el diámetro interior de los ventrículos, cuando se compara con los ratones sanos. Los datos sugieren que los cambios morfológicos y funcionales en los corazones inflamados pueden ser de forma no invasiva monitoreado por MRM en animales vivos. En conclusión, MRM ofrece una ventaja de la evaluación de la progresión y regresión de las lesiones del miocardio en enfermedades causadas por agentes infecciosos, así como la respuesta a las terapias.

En este informe, se muestra la utilidad de la técnica de MRM como una modalidad no invasiva para determinar los cambios estructurales y funcionales en el corazón de los animales afectados con EAM. La miocarditis se indujo en ratones A / J mediante la inmunización de los animales con MyHC-α 334-352 en CFA 7, y los animales se sometieron a experimentos de GRM en el día 21 después de la inmunización. La proyección de imagen de MRM se realizó en animales vivos bajo anestesia con isoflurano al 9,4 T (400 ...

Watch this Scientific Journal Video about Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse at JoVE.com

Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

Evaluación no invasiva de la cardiaca Anomalías en la miocarditis autoinmune experimental por Resonancia Magnética de imágenes de microscopía en el ratón

Myocarditis is an inflammation of the myocardium, but only ~10% of those affected show clinical manifestations of the disease. To study the immune events ...

noninvasive-assessment-cardiac-abnormalities-experimental-autoimmune

Research

JoVE Journal

Medicine

9.8K vistas.  University of Nebraska-Lincoln. This study demonstrates the successful establishment of magnetic resonance microscopy imaging as a non-invasive tool to assess the cardiac abnormalities in mice affected with autoimmune myocarditis. The data indicate that the technique can be used to monitor the disease-progression in live animals.

Video: Noninvasive Assessment of Cardiac Abnormalities in Experimental Autoimmune Myocarditis by Magnetic Resonance Microscopy Imaging in the Mouse

Magnetic Resonance Derived Myocardial Strain Assessment Using Feature Tracking

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF. Current technologies for CMR are time consuming and difficult to implement in clinical practice. Feature tracking is a technology that can lead to more automization and robustness of quantitative analysis of medical images with less time consumption than comparable methods.
Methods: An automatic or manual input in a single phase serves as an initialization from which the system starts to track the displacement of individual patterns representing anatomical structures over time. The specialty of this method is that the images do not need to be manipulated in any way beforehand like e.g. tagging of CMR images.
Results: The method is very well suited for tracking muscular tissue and with this allowing quantitative elaboration of myocardium and also blood flow.
Conclusions: This new method offers a robust and time saving procedure to quantify myocardial tissue and blood with displacement, velocity and deformation parameters on regular sequences of CMR imaging. It therefore can be implemented in clinical practice.

An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that strain is a more sensitive and earlier marker for contractile dysfunction than the frequently used parameter EF.

Evaluación de Resonancia Magnética Derivado deformación miocárdica mediante función de seguimiento

Purpose: An accurate and practical method to measure parameters like strain in myocardial tissue is of great clinical value, since it has been shown, that ...

Investigación

Medicina

Multi-modal Imaging of Angiogenesis in a Nude Rat Model of Breast Cancer Bone Metastasis Using Magnetic Resonance Imaging, Volumetric Computed Tomography and Ultrasound

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell proliferation in the bone marrow cavity as well as for interaction of tumor and bone cells resulting in local bone destruction. Our aim was to develop a model of experimental bone metastasis that allows in vivo assessment of angiogenesis in skeletal lesions using non-invasive imaging techniques.
For this purpose, we injected 105 MDA-MB-231 human breast cancer cells into the superficial epigastric artery, which precludes the growth of metastases in body areas other than the respective hind leg1. Following 25-30 days after tumor cell inoculation, site-specific bone metastases develop, restricted to the distal femur, proximal tibia and proximal fibula1. Morphological and functional aspects of angiogenesis can be investigated longitudinally in bone metastases using magnetic resonance imaging (MRI), volumetric computed tomography (VCT) and ultrasound (US).
MRI displays morphologic information on the soft tissue part of bone metastases that is initially confined to the bone marrow cavity and subsequently exceeds cortical bone while progressing. Using dynamic contrast-enhanced MRI (DCE-MRI) functional data including regional blood volume, perfusion and vessel permeability can be obtained and quantified2-4. Bone destruction is captured in high resolution using morphological VCT imaging. Complementary to MRI findings, osteolytic lesions can be located adjacent to sites of intramedullary tumor growth. After contrast agent application, VCT angiography reveals the macrovessel architecture in bone metastases in high resolution, and DCE-VCT enables insight in the microcirculation of these lesions5,6. US is applicable to assess morphological and functional features from skeletal lesions due to local osteolysis of cortical bone. Using B-mode and Doppler techniques, structure and perfusion of the soft tissue metastases can be evaluated, respectively. DCE-US allows for real-time imaging of vascularization in bone metastases after injection of microbubbles7.
In conclusion, in a model of site-specific breast cancer bone metastases multi-modal imaging techniques including MRI, VCT and US offer complementary information on morphology and functional parameters of angiogenesis in these skeletal lesions.

In the pathogenesis of bone metastasis, angiogenesis is a crucial process and therefore represents a target for imaging and therapy. Here, we present a rat model of site-specific breast cancer bone metastasis and describe strategies to non-invasively image angiogenesis in vivo using magnetic resonance imaging, volumetric computed tomography and ultrasound.

Multimodal de imágenes de la angiogénesis en un modelo de rata desnuda de las metástasis óseas del cáncer de pecho utilizando imágenes de resonancia magnética, tomografía computarizada y la ecografía volumétrica

Angiogenesis is an essential feature of cancer growth and metastasis formation. In bone metastasis, angiogenic factors are pivotal for tumor cell ...

Diffusion Tensor Magnetic Resonance Imaging in the Analysis of Neurodegenerative Diseases

Diffusion tensor imaging (DTI) techniques provide information on the microstructural processes of the cerebral white matter (WM) in vivo. The present applications are designed to investigate differences of WM involvement patterns in different brain diseases, especially neurodegenerative disorders, by use of different DTI analyses in comparison with matched controls. 	
DTI data analysis is performed in a variate fashion, i.e. voxelwise comparison of regional diffusion direction-based metrics such as fractional anisotropy (FA), together with fiber tracking (FT) accompanied by tractwise fractional anisotropy statistics (TFAS) at the group level in order to identify differences in FA along WM structures, aiming at the definition of regional patterns of WM alterations at the group level. Transformation into a stereotaxic standard space is a prerequisite for group studies and requires thorough data processing to preserve directional inter-dependencies. The present applications show optimized technical approaches for this preservation of quantitative and directional information during spatial normalization in data analyses at the group level. On this basis, FT techniques can be applied to group averaged data in order to quantify metrics information as defined by FT. Additionally, application of DTI methods, i.e. differences in FA-maps after stereotaxic alignment, in a longitudinal analysis at an individual subject basis reveal information about the progression of neurological disorders. Further quality improvement of DTI based results can be obtained during preprocessing by application of a controlled elimination of gradient directions with high noise levels.	
In summary, DTI is used to define a distinct WM pathoanatomy of different brain diseases by the combination of whole brain-based and tract-based DTI analysis.

Diffusion tensor imaging (DTI) basically serves as an MRI-based tool to identify in vivo the microstructure of the brain and pathological processes due to neurological disorders within the cerebral white matter. DTI-based analyses allow for application to brain diseases both at the group level and in single subject data.

Tensor de difusión por resonancia magnética en el análisis de las enfermedades neurodegenerativas

Diffusion tensor imaging (DTI) techniques provide information on the microstructural processes of the cerebral white matter (WM) in vivo. The present ...

Implanting Glass Spinal Cord Windows in Adult Mice with Experimental Autoimmune Encephalomyelitis

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as multiple sclerosis. Here we present a low-cost and reproducible glass window implantation protocol that is suitable for intravital microscopy and studying the dynamics of spinal cord cytoarchitecture with subcellular resolution in live adult mice with EAE. Briefly, we surgically expose the vertebrae T12-L2 and construct a chamber around the exposed vertebrae using a combination of cyanoacrylate and dental cement. A laminectomy is performed from T13 to L1, and a thin layer of transparent silicone elastomer is applied to the dorsal surface of the exposed spinal cord. A modified glass cover slip is implanted over the exposed spinal cord taking care that the glass does not directly contact the spinal cord. To reduce the infiltration of inflammatory cells between the window and spinal cord, anti-inflammatory treatment is administered every 2 days (as recommended by ethics committee) for the first 10 days after implantation. EAE is induced only 2-3 weeks after the cessation of anti-inflammatory treatment.
Using this approach we successfully induced EAE in 87% of animals with implanted windows and, using Thy1-CFP-23 mice (blue axons in dorsal spinal cord), quantified axonal loss throughout EAE progression. Taken together, this protocol may be useful for studying the recruitment of various cell populations as well as their interaction dynamics, with subcellular resolution and for extended periods of time. This intravital imaging modality represents a valuable tool for developing therapeutic strategies to treat autoimmune demyelinating diseases such as EAE.

We describe a method for implanting and maintaining glass windows over the exposed spinal cords of adult mice for studying experimental autoimmune encephalomyelitis. These windows provide chronic optical access to the spinal cord for monitoring cell dynamics at the subcellular level in live animals using two-photon microscopy.

Experimental autoimmune encephalomyelitis (EAE) in adult rodents is the standard experimental model for studying autonomic demyelinating diseases such as ...

Simultaneous PET/MRI Imaging During Mouse Cerebral Hypoxia-ischemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics disturbance in affected cells. Diffusion weighted magnetic resonance imaging (MRI) identifies regions that are damaged, potentially irreversibly, by hypoxia-ischemia. Alterations in glucose utilization in the affected tissue may be detectable by positron emission tomography (PET) imaging of 2-deoxy-2-(18F)fluoro-&#7429;-glucose ([18F]FDG) uptake. Due to the rapid and variable nature of injury in this animal model, acquisition of both modes of data must be performed simultaneously in order to meaningfully correlate PET and MRI data. In addition, inter-animal variability in the hypoxic-ischemic injury due to vascular differences limits the ability to analyze multi-modal data and observe changes to a group-wise approach if data is not acquired simultaneously in individual subjects. The method presented here allows one to acquire both diffusion-weighted MRI and [18F]FDG uptake data in the same animal before, during, and after the hypoxic challenge in order to interrogate immediate physiological changes.

The method presented here uses simultaneous positron emission tomography and magnetic resonance imaging. In the cerebral hypoxia-ischemia model, dynamic changes in diffusion and glucose metabolism occur during and after injury. The evolving and irreproducible damage in this model necessitates simultaneous acquisition if meaningful multi-modal imaging data are to be acquired.

Simultánea de imágenes PET / RM Durante cerebral del ratón hipoxia-isquemia

Dynamic changes in tissue water diffusion and glucose metabolism occur during and after hypoxia in cerebral hypoxia-ischemia reflecting a bioenergetics ...

Assessment of Cardiac Morphological and Functional Changes in Mouse Model of Transverse Aortic Constriction by Echocardiographic Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable noninvasive method is essential to assess real-time cardiac morphological and functional changes in animal models of heart disease. Transthoracic echocardiography represents an important tool for noninvasive assessment of cardiac structure and function2. Here we used a high-resolution ultrasound imaging system to monitor myocardial remodeling and heart failure progression over time in a mouse model of TAC. B-mode, M-mode, and Doppler imaging were used to precisely assess cardiac hypertrophy, ventricular dilatation, and functional deterioration in mice following TAC. Color and pulse wave (PW) Doppler imaging was used to noninvasively measure pressure gradient across the aortic constriction created by TAC and to assess transmitral blood flow in mice. Thus transthoracic echocardiographic imaging provides comprehensive noninvasive measurements of cardiac dimensions and function in mouse models of heart disease.

The goal of this protocol is to noninvasively assess cardiac structural and functional changes in a mouse model of heart disease created by transverse aortic constriction, using B- and M-mode echocardiography and color/pulse wave Doppler imaging.

Evaluación de la cardíaco morfológica y funcional Los cambios en el modelo de ratón de la constricción de la aorta transversa por ecocardiográfico Imaging

Transverse aortic constriction (TAC) in mice has been used as a valuable model to study mechanisms of cardiac hypertrophy and heart failure1. A reliable ...

Quantitative Visualization of Leukocyte Infiltrate in a Murine Model of Fulminant Myocarditis by Light Sheet Microscopy

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently labelled structures in large biological specimens, and is down to cellular resolution. Meanwhile, the constant refinement of underlying protocols and the enhanced availability of specialized commercial systems enable us to investigate the microstructure of whole mouse organs and even allow for the characterization of cellular behavior in various live-cell imaging approaches. Here, we describe a protocol for the spatial whole-mount visualization and quantification of the CD45+ leukocyte population in inflamed mouse hearts. The method employs a transgenic mouse strain (CD11c.DTR)that has recently been shown to serve as a robust, inducible model for the study of the development of fulminant fatal myocarditis, characterized by lethal cardiac arrhythmias. This protocol includes myocarditis induction, intravital antibody-mediated cell staining, organ preparation, and LSFM with subsequent computer-assisted image post-processing. Although presented as a highly-adapted method for our particular scientific question, the protocol represents the blueprint of an easily adjustable system that can also target completely different fluorescent structures in other organs and even in other species.

Here, we describe a light-sheet microscopy approach to visualize the cardiac CD45+ leukocyte infiltrate in a murine model of aseptic fulminant myocarditis, which is induced by the intratracheal diphtheria toxin treatment of CD11c.DTR mice.

Visualización Cuantitativa del Infiltrado de Leucocitos en un Modelo Murino de Miocarditis Fulminante por Microscopía de Hoja Ligera

Light-sheet fluorescence microscopy (LSFM), in combination with chemical clearing protocols, has become the gold standard for analyzing fluorescently ...

Cardiac Magnetic Resonance Imaging at 7 Tesla

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher magnetic field strengths and potentially provides improved signal contrast and spatial resolution. While promising results have been achieved, ultra-high field CMR is challenging due to energy deposition constraints and physical phenomena such as transmission field non-uniformities and magnetic field inhomogeneities. In addition, the magneto-hydrodynamic effect renders the synchronization of the data acquisition with the cardiac motion difficult. The challenges are currently addressed by explorations into novel magnetic resonance technology. If all impediments can be overcome, ultra-high field CMR may generate new opportunities for functional CMR, myocardial tissue characterization, microstructure imaging or metabolic imaging. Recognizing this potential, we show that multi-channel radio frequency (RF) coil technology tailored for CMR at 7 Tesla together with higher order B0 shimming and a backup signal for cardiac triggering facilitates high fidelity functional CMR. With the proposed setup, cardiac chamber quantification can be accomplished in examination times similar to those achieved at lower field strengths. To share this experience and to support the dissemination of this expertise, this work describes our setup and protocol tailored for functional CMR at 7 Tesla.

The sensitivity gain inherent to ultrahigh field magnetic resonance holds promise for high spatial resolution imaging of the heart. Here, we describe a protocol customized for functional cardiovascular magnetic resonance (CMR) at 7 Tesla using an advanced multi-channel radio-frequency coil, magnetic field shimming and a triggering concept.

Resonancia magnética cardiaca en 7 Tesla

CMR at an ultra-high field (magnetic field strength B0&#160;&#8805; 7 Tesla) benefits from the signal-to-noise ratio (SNR) advantage inherent at higher ...

Phase Contrast Magnetic Resonance Imaging in the Rat Common Carotid Artery

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous studies have shown that abnormal blood flow may be associated with systemic vascular risk. Thus, PC-MRI can facilitate the translation of data obtained from animal models of cardiovascular diseases to pertinent clinical investigations. In this report, we describe the procedure for measuring blood flow in the common carotid artery (CCA) of rats using cine-gated PC-MRI and discuss relevant analysis methods. This procedure can be performed in a live, anesthetized animal and does not require euthanasia after the procedure. The proposed scanning parameters yield repeatable measurements for blood flow, indicating excellent reproducibility of the results. The PC-MRI procedure described in this article can be used for pharmacological testing, pathophysiological assessment, and cerebral hemodynamics evaluation.

The overall goal of this procedure is to measure the blood flow in the rat common carotid artery by using noninvasive phase contrast magnetic resonance imaging.

Fase contraste resonancia magnética en la arteria carótida común de rata

Phase contrast magnetic resonance imaging (PC-MRI) is a noninvasive approach that can quantify flow-related parameters such as blood flow. Previous ...

Quantification of Levator Ani Hiatus Enlargement by Magnetic Resonance Imaging in Males and Females with Pelvic Organ Prolapse

Here we present a protocol to examine the levator ani hiatus in males and females with pelvic organ prolapse, during the Valsalva maneuver and while evacuating acoustic gel, using a horizontally oriented 1.5 T magnetic resonance (MR) scanner. On midsagittal images, the vertical distance of pelvic organs is measured in millimeters relative to the hymen plane (female) and to the lower border of the symphysis pubis (male), preceded by - (above) or + ( below) signs. On axial images, the levator ani area is calculated in square centimeters with a free-hand tracing method from three key images, passing through the midsymphysis (level I), tangential to the lower border of the symphysis (level II), and at the maximal anterior rectal wall bulging (level III). Areas at rest and strained are compared to find evidence of a percentage of increase. The purpose is to provide objective evidence of the maximal extent of pelvic organs descent and hiatus enlargement without the interference of foreign objects or the examiner&#39;s proximity, so as to overcome the limitations of pelvic examination and transperineal sonography (i.e., subjectivity and sex-related limitations [female only]).

Here we present a protocol to standardize the measurement of the levator ani hiatus size&#160;by magnetic resonance imaging. The purpose is to extract biomechanical inferences from image analysis by comparing resting and straining values in patients of both sexes with pelvic prolapse, using consistent anatomical bony landmarks.

Cuantificación de la ampliación de hiato de Ani del Levator por resonancia magnética en hombres y mujeres con prolapso de órganos pélvicos

Here we present a protocol to examine the levator ani hiatus in males and females with pelvic organ prolapse, during the Valsalva maneuver and while ...