This project was supported by Grant Numbers HL96593 from NIH and D56HP20783 from HRSA/ HHS. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH or HRSA / HHS.

Nota: El proceso de adquisición para la obtención de imágenes capilares ya ha sido publicado y se logra utilizando una cámara fotográfica digital con un programa informático de adquisición y análisis de imágenes correspondiente 11 21 Esta práctica de laboratorio utiliza imágenes fijas para el análisis, no vídeos, lo que simplifica la adquisición de imágenes para el análisis.. A continuación se describe la nueva técnica para cuantificar los capilares a partir de las imágenes. 1. Mejora de la imagen de proceso <ol><li> Obtener imágenes digitales con una cámara digital monocromo. Calibrar las imágenes a un objeto de tamaño conocido por tomar una foto de un objeto de longitud conocida como una regla con la cámara. Este proceso permite que el programa informático para medir y contar capilares con precisión después del procesamiento. Lo ideal para la más alta precisión, se debe utilizar una retícula (pieza científicamente fabricado de vidrio con una regla grabada en ella). Medir el número de píxeles en un cuadrado de 1 mmcaja usando un programa informático. Nota: La clave para la reproducibilidad y la normalización de este protocolo depende en gran medida de la correcta colocación de la caja 1 mm 2 que se cuenta. </li><li> Utilice herramientas de mejora de contraste para oscurecer los capilares y aclarar el fondo, lo que maximizará la visualización de los capilares. La diferenciación inicial de los capilares de fondo es importante para el cultivo adecuado de la imagen en pasos posteriores. Para ello, el ajuste de la imagen mediante un Para ello haga clic en &quot;mejor histograma en forma.&quot;: La captura, la intensidad, la imagen de histograma, mejor ajuste. </li><li> Recortar una región de interés (ROI) para el recuento capilar como una nueva imagen (seleccionar, regiones de interés). Utilice una caja de 1 mm 2, que se determinó mediante la calibración de las imágenes a 530 píxeles igualando 1 mm. compruebe que la imagen recortada coloca el ápice de las asas capilares en la parte superior de la imagen. </li><li> Aplanar la imagen para que todos los ajustes de imagen futuras se aplicarán uniformemente ala imagen. Para ello, haga clic en: Ficha proceso, filtros 2D, aplanar, intensidad BG de &quot;brillante&quot; object width a &quot;75&quot;, se aplica. </li><li> El contraste de la imagen para los capilares se visualizan al máximo. Para ello, haga clic en: ficha, pantalla ajustar, aumentar contraste con 75. </li><li> Eliminar manchas en la imagen para suavizar los bordes de los capilares haciendo clic en la pestaña de procesos, filtros 2D, despeckle, el tamaño del núcleo 7 x 7, aplique. </li><li> Finalizar el contraste de la imagen de modo capilares son de color negro y el fondo es blanco. Realice este paso ajustando el histograma para el modelo de &quot;mejor ajuste&quot;. Nota: Por favor consulte la Figura 2 en resultados representativos para un ejemplo de lo que la imagen procesada debe ser similar siguiendo estos pasos. </li></ol> 2. Realizar Condes capilares / cuantificar capilar Densidad <ol><li> En cada imagen, seleccionar manualmente una parte de un capilar bien definida mediante el featur &quot;objeto destino&quot;e para ser reconocido como objetos a ser contados por el programa. A continuación, seleccione una pequeña parte del fondo, utilizando la función de &quot;fondo&quot;, como referencia a las áreas que necesitan ser ignorado por el algoritmo de recuento. Nota: La combinación de estos destacados provoca todos los capilares a destacar sin tener en cuenta el ruido de fondo. El protocolo de cuantificación (contando) utiliza las funciones del ordenador para diferenciar partes de la imagen basada en el color y la morfología. </li><li> Utilice la función de recuento para contar al instante todos los capilares en la imagen con el equipo de imagen. Ajuste el diámetro mínimo de los objetos contados a 5 píxeles con el fin de evitar contar el ruido de fondo como capilares. </li><li> Para cada individuo, contar el promedio de 3 - 4 imágenes con el fin de obtener una evaluación más fiable. Nota: Por favor consulte la Figura 3 en resultados representativos para un ejemplo de lo que la imagen debe ser similar durante el procedimiento de recuento. </li></ol&gt; 3. Creación y uso de macros para automatizar Procesamiento de Imágenes Nota: Para ahorrar tiempo, las macros se pueden crear con el fin de realizar automáticamente una secuencia específica de los procesos en una o varias imágenes. Estas secuencias pueden ser personalizados con el fin de hacer las modificaciones de imagen más rápida. En esencia, estas macros recuerdan cómo se procesan las imágenes, y llevar a cabo todos los pasos de forma rápida y sin intervención del usuario. Realización de los recuentos en 12 imágenes capilares toma este laboratorio entre 20 a 30 min con las macros (2 a 3 min por imagen), en oposición a aproximadamente 8 min por imagen sin los macros. Por lo tanto el uso de las macros es de 3 a 5 veces más eficiente que el procesamiento manualmente cada imagen individual. <ol><li> Con el fin de crear una macro, seleccione &quot;Grabar macro&quot; y en una imagen de realizar los pasos y procesos deseados, como se describe en los pasos 1 y 2 anteriores. Nombre de la macro a partir del cual la imagen pasos de procesamiento se realizaron para referencia futura. Cuando using la macro en futuras imágenes, simplemente haga clic en &quot;macro run&quot; y el programa se aplicará automáticamente las mejoras registradas a la imagen (s) deseada. Nota: Este laboratorio utiliza una macro para realizar todos menos uno de los pasos en la Sección 1 de los métodos en unos segundos. El único paso que requiere la entrada del usuario es la elección de dónde recortar la imagen, paso 1.2. </li></ol>

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The authors have no conflicts of interest.

Capilaroscopia periungueal se muestra prometedor como una herramienta clínicamente útil en el futuro para diversos oncología, cardiovascular, y las aplicaciones de enfermedades reumatológicas. El proceso de adquisición de imágenes es bastante consistente entre los investigadores, sin embargo, en la actualidad hay varios métodos para el procesamiento de imágenes y análisis. Métodos actualmente incluyen recuentos capilares computarizados y manuales. Conteos manuales son problemáticos, ya que son mucho tiempo, y...

Formación de imágenes microvascular es un campo en rápido crecimiento con muchas aplicaciones clínicas potenciales. 2 Por ejemplo, los oncólogos están utilizando formación de imágenes de los microvasos para determinar la extensión de la angiogénesis tumoral, produciendo una valiosa información sobre el estado del tumor y una visión de posibles opciones de tratamiento. 3 4 Sin embargo, capilaroscopia lecho ungueal es quizás la forma eficiente y ampliamente aplicable costo mayor parte de las imágenes microvascular. Los investigadores están utilizando el vídeo lecho ungueal capilaroscopia para estudiar las tasas de flujo de sangre e investigar la morfología capilar. 5 6 Tanto vídeo y de imágenes fijas lecho ungueal capilaroscopia están adjuntos a la atención para diagnosticar y tratar fenómeno y varias enfermedades del tejido conectivo de Raynaud, como la esclerosis sistémica. 2 Capilaroscopia periungueal tiene diversas aplicaciones cardiovasculares potenciales. La investigación actual sugiere usar lecho ungueal capilaroscopiaEse tipo diabetes mellitus 1 y Tipo 2 pacientes presentan una alta prevalencia de la morfología capilar anormal, sin embargo, tienen densidades capilares sin cambios en comparación con los individuos no diabéticos. 8.7 capilaroscopia también se ha estudiado experimentalmente en la hipertensión. Estructural rarefacción capilar que conduce a una reducción de la densidad capilar se ha demostrado en individuos hipertensos en comparación con los individuos no hipertensos. 9-10 En contraste con estos pacientes hipertensos de edad avanzada que presentan rarefacción estructurales, la investigación más reciente ha demostrado que (edad 40 y por encima de la media) pacientes hipertensos más jóvenes (media de edad menores de 40 años) tienen rarefacción funcional sin rarefacción estructural. 11 Esto sugiere que la rarefacción funcional se produce antes y puede progresar con el tiempo para rarefacción estructural. Curiosamente, los pacientes hipertensos tratados con fármacos antihipertensivos específicos como perindopril / indapamida representada normaAl densidad capilar y la función endotelial después del tratamiento, mientras que los tratados con ACE (Inhibidores de la enzima convertidora de) inhibidores o diuréticos mantiene una baja densidad capilar a pesar del control de la presión arterial comparable. 12 Esto sugiere que algunos medicamentos antihipertensivos pueden normalizar la densidad capilar invirtiendo el capilar rarefacción causado por la hipertensión. Además, otros investigadores han demostrado que una reducción en la ingesta de sal conduce a la reversión de tanto rarefacción capilar funcional y estructural en individuos hipertensos. 13 A pesar de las diversas aplicaciones clínicas potenciales de esta tecnología, hay poca estandarización de la técnica para la cuantificación de las imágenes de densidad capilar. 2 Hasta la fecha, los investigadores han encontrado que los resultados de densidad capilar son reproducibles, tanto desde el intra-observador y la perspectiva inter-observador sólo si la exacta misma área está siendo contado cada vez. 1,14 15 </sup&gt; Es de destacar que los investigadores anteriores han realizado gran parte recuentos capilares manualmente con el ojo desnudo, 9 16 17 18, que es un proceso lento y subjetiva. Algoritmos estandarizados, basados ​​informáticos para la cuantificación de las imágenes capilares teóricamente proporcionan un análisis de datos más eficiente y reproducible con menos subjetividad, lo que facilita las aplicaciones clínicas de capilaroscopia. Algunos investigadores han utilizado efectivamente los programas basados ​​en computadoras para cuantificar los datos de nailfold imágenes capilaroscópicos. 1,6 19 20 Sin embargo, sólo una publicación a la fecha describe fiabilidad de un programa de software complejo disponibles para cuantificar los datos capilaroscopia, 1 y este programa es complicado como anteriormente se ha indicado anteriormente por el requisito de contar el mismo campo visual exacto. A continuación, presentamos un protocolo sencillo y fiable para cuantificar los capilares utilizando un algoritmo estandarizado que permiteel uso de múltiples campos visuales. El uso de múltiples campos visuales no sólo simplifica el procedimiento, sino que también permite la evaluación de la variación biológica normal en el recuento capilar. El objetivo de este estudio es describir un algoritmo computarizado reproducible y eficiente, que estandariza el proceso de cuantificación capilar. Si bien estos métodos no están completamente automatizados que requieren muy poca intervención del usuario, y proporcionan la cuantificación rápida y fiable de las imágenes.

<table>
	<tbody>
		<tr>
			<td class="xl65">Image-Pro Premier</td>
			<td class="xl65">Media Cybernetics, Inc</td>
			<td align="right" class="xl63">9.1</td>
			<td class="xl63">Image processing software</td>
		</tr>
	</tbody>
</table>

a reproducible computerized method for quantitation of capillary density using nailfold capillaroscopy

Capilaroscopia es una metodología no invasiva, eficiente, relativamente barato y fácil de aprender para visualizar directamente la microcirculación. La técnica capilaroscopia puede proporcionar información sobre la salud de un paciente microvascular, lo que lleva a una variedad de dermatológica potencialmente valiosa, oftalmológica, aplicaciones clínicas reumatológicas y cardiovasculares. Además, el crecimiento del tumor puede ser dependiente de la angiogénesis, que puede ser cuantificó midiendo la densidad de microvasos dentro del tumor. Sin embargo, en la actualidad existe poca o ninguna estandarización de las técnicas, y sólo una publicación a la fecha reporta la fiabilidad de unas actualmente disponibles algoritmos basados ​​ordenador y complejas para cuantificar los datos capilaroscopia. 1 En este trabajo se describe un nuevo, más simple,, algoritmo fiable recuento capilar estandarizado para cuantificar los datos capilaroscopia nailfold. Un algoritmo capilaroscopia computarizada simple, reproducible como esto facilitaría másuso generalizado de la técnica entre los investigadores y los clínicos. Muchos investigadores analizan actualmente imágenes capilaroscopia a mano, la promoción de la fatiga del usuario y de la subjetividad de los resultados. Este documento describe un nuevo, fácil de uso automatizado algoritmo de procesamiento de imágenes, además de un algoritmo de conteo semi-automatizado reproducible. Este algoritmo permite el análisis de las imágenes en cuestión de minutos, mientras que la reducción de la subjetividad; sólo una cantidad mínima de tiempo de entrenamiento (en nuestra experiencia, a menos de 1 hora) es necesario para aprender la técnica.

El objetivo de este procedimiento de procesamiento de imágenes es diferenciar los capilares a partir de la imagen de fondo para que puedan ser cuantificados con precisión. Tanto el procesamiento de imágenes y procesamiento de imágenes incompleta excesiva son perjudiciales para la capacidad del programa para cuantificar los capilares. Como se ve en la Figura 3, el procesamiento de imágenes hace que los capilares incompleta difícil de distinguir del fondo. Es crítico que el usuario...

Watch this Scientific Journal Video about A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy at JoVE.com

A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy

Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy-to-learn methodology for directly visualizing capillaries in the microcirculation. However, only one publication to date describes the reliability of a complex software program available for quantitating capillaroscopy data. Here, we present a simple, reliable protocol for quantitating capillaries using a standardized algorithm.

Un método computarizado reproducible para la cuantificación de la densidad capilar utilizando periungueal capilaroscopia

Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy to learn methodology for directly visualizing the microcirculation. The ...

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Research

JoVE Journal

Medicine

8.9K vistas.  Thomas Jefferson University. Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy-to-learn methodology for directly visualizing capillaries in the microcirculation. However, only one publication to date describes the reliability of a complex software program available for quantitating capillaroscopy data. Here, we present a simple, reliable protocol for quantitating capillaries using a standardized algorithm. 

Video: A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy

Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected light-sensitive dye. Following illumination, the dye is activated and produces singlet oxygen that damages components of endothelial cell membranes, with subsequent platelet aggregation and thrombi formation, which eventually determines the interruption of local blood flow. This approach, initially proposed by Rosenblum and El-Sabban in 1977, was later improved by Watson in 1985 in rat brain and set the basis of the current model. Also, the increased availability of transgenic mouse lines further contributed to raise the interest on the photothrombosis model. Briefly, a photosensitive dye (Rose Bengal) is injected intraperitoneally and enters the blood stream. When illuminated by a cold light source, the dye becomes activated and induces endothelial damage with platelet activation and thrombosis, resulting in local blood flow interruption. The light source can be applied on the intact skull with no need of craniotomy, which allows targeting of any cortical area of interest in a reproducible and non-invasive way. The mouse is then sutured and allowed to wake up. The evaluation of ischemic damage can be quickly accomplished by triphenyl-tetrazolium chloride or cresyl violet staining. This technique produces infarction of small size and well-delimited boundaries, which is highly advantageous for precise cell characterization or functional studies. Furthermore, it is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombosis is a quick, minimally-invasive technique for inducing small and well-delimited infarction in areas of interest in highly reproducible manner. It is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Isquemia photothrombotic: Un modelo de lesión cortical fotoquímico mínimamente invasiva y reproducible de Estudios Carrera Ratón

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected ...

Investigación

Medicina

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating important information on the pathogenesis of lesion formation and the action of novel therapies. Use of atherosclerosis-prone mice, surgical methods of lesion induction, and dietary modification has dramatically improved understanding of the mechanisms that contribute to disease development and the potential of new treatments.
Classically, analysis of lesions is performed ex vivo using 2-dimensional histological techniques. This article describes application of optical projection tomography (OPT) to 3-dimensional quantitation of arterial lesions. As this technique is non-destructive, it can be used as an adjunct to standard histological and immunohistochemical analyses.
Neointimal lesions were induced by wire-insertion or ligation of the mouse femoral artery whilst atherosclerotic lesions were generated by administration of an atherogenic diet to apoE-deficient mice.
Lesions were examined using OPT imaging of autofluorescent emission followed by complementary histological and immunohistochemical analysis. OPT clearly distinguished lesions from the underlying vascular wall. Lesion size was calculated in 2-dimensional sections using planimetry, enabling calculation of lesion volume and maximal cross-sectional area. Data generated using OPT were consistent with measurements obtained using histology, confirming the accuracy of the technique and its potential as a complement (rather than alternative) to traditional methods of analysis.
This work demonstrates the potential of OPT for imaging atherosclerotic and neointimal lesions. It provides a rapid, much needed ex vivo technique for the routine 3-dimensional quantification of vascular remodelling.

Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.

Generación y cuantificación de 3 dimensiones de las lesiones arteriales en ratones mediante tomografía óptica de proyección

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Computerized Dynamic Posturography for Postural Control Assessment in Patients with Intermittent Claudication

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic conditions. As part of a diagnostic assessment, the early detection of postural deficits is important so that appropriate and targeted interventions can be prescribed. The Sensory Organization Test (SOT) on the EquiTest determines an individual&#39;s use of the sensory systems (somatosensory, visual, and vestibular) that are responsible for postural control. Somatosensory and visual input are altered by the calibrated sway-referenced support surface and visual surround, which move in the anterior-posterior direction in response to the individual&#39;s postural sway. This creates a conflicting sensory experience. The Motor Control Test (MCT) challenges postural control by creating unexpected postural disturbances in the form of backwards and forwards translations. The translations are graded in magnitude and the time to recover from the perturbation is computed.
Intermittent claudication, the most common symptom of peripheral arterial disease, is characterized by a cramping pain in the lower limbs and caused by muscle ischemia secondary to reduced blood flow to working muscles during physical exertion. Claudicants often display poor balance, making them susceptible to falls and activity avoidance. The Ankle Brachial Pressure Index (ABPI) is a noninvasive method for indicating the presence of peripheral arterial disease and intermittent claudication, a common symptom in the lower extremities. ABPI is measured as the highest systolic pressure from either the dorsalis pedis or posterior tibial artery divided by the highest brachial artery systolic pressure from either arm. This paper will focus on the use of computerized dynamic posturography in the assessment of balance in claudicants.

Individuals with intermittent claudication exhibit poor balance compared to healthy controls. Computerized dynamic posturography is an objective method for measuring an individual's postural responses to balance disturbances. This provides an objective reflection of the person&#x2019;s ability to respond to situations under which sensory stimuli are altered and unexpected perturbations occur.

Posturografía dinámica computarizada para la evaluación de control postural en pacientes con claudicación intermitente

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic ...

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States. Mortality is due to diagnosis of 75% of women with late stage disease, when metastasis is already present. EOC is characterized by diffuse and widely disseminated intra-peritoneal metastasis. Cells shed from the primary tumor anchor in the mesothelium that lines the peritoneal cavity as well as in the omentum, resulting in multi-focal metastasis, often in the presence of peritoneal ascites. Efforts in our laboratory are directed at a more detailed understanding of factors that regulate EOC metastatic success. However, quantifying metastatic tumor burden represents a significant technical challenge due to the large number, small size and broad distribution of lesions throughout the peritoneum. Herein we describe a method for analysis of EOC metastasis using cells labeled with red fluorescent protein (RFP) coupled with in vivo multispectral imaging. Following intra-peritoneal injection of RFP-labelled tumor cells, mice are imaged weekly until time of sacrifice. At this time, the peritoneal cavity is surgically exposed and organs are imaged in situ. Dissected organs are then placed on a labeled transparent template and imaged ex vivo. Removal of tissue auto-fluorescence during image processing using multispectral unmixing enables accurate quantitation of relative tumor burden. This method has utility in a variety of applications including therapeutic studies to evaluate compounds that may inhibit metastasis and thereby improve overall survival.

Ovarian cancer metastasis is characterized by numerous diffuse intra-peritoneal lesions, such that accurate visual quantitation of tumor burden is challenging. Herein we describe a method for in situ and ex vivo quantitation of metastatic tumor burden using red fluorescent protein (RFP)-labeled tumor cells and optical imaging.

La cuantificación de la intra-peritoneal del cáncer ovárico Metástasis

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

Método de la entrega directa segmentaria intra-hepática utilizando un modelo de hígado de rata hiliar abrazadera

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

Reproducible lesión arterial Denudación por pinzamiento aórtico abdominal infrarrenal en un modelo murino

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

A Method for Quantifying Upper Limb Performance in Daily Life Using Accelerometers

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. It has become important to measure a person's activities in daily life, and not just measure their capacity for activity in the structured environment of a clinic or laboratory. A wearable sensor that is now enabling measurement of daily movement is the accelerometer. Accelerometers are commercially-available devices resembling large wrist watches that can be worn throughout the day. Data from accelerometers can quantify how the limbs are engaged to perform activities in peoples' homes and communities. This report describes a methodology to collect accelerometry data and turn it into clinically-relevant information. First, data are collected by having the participant wear two accelerometers (one on each wrist) for 24 h or longer. The accelerometry data are then downloaded and processed to produce four different variables that describe key aspects of upper limb activity in daily life: hours of use, use ratio, magnitude ratio, and the bilateral magnitude. Density plots can be constructed that visually represent the data from the 24 h wearing period. The variables and their resultant density plots are highly consistent in neurologically-intact, community-dwelling adults. This striking consistency makes them a useful tool for determining if upper limb daily performance is different from normal. This methodology is appropriate for research studies investigating upper limb dysfunction and interventions designed to improve upper limb performance in daily life in people with stroke and other patient populations. Because of its relative simplicity, it may not be long before it is also incorporated in clinical neurorehabilitation practice.

This protocol describes a method to quantify upper limb performance in daily life using wrist-worn accelerometers.

Un método para cuantificar Rendimiento de las extremidades superiores en la Vida Diaria El uso de acelerómetros

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. ...

Ablation of Ischemic Ventricular Tachycardia Using a Multipolar Catheter and 3-dimensional Mapping System for High-density Electro-anatomical Reconstruction

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars represent zones of slow conduction that allow the occurrence and maintenance of reentrant circuits. Catheter ablation enables substrate modification of those low voltage areas and thus can help to alter the scar tissue in such a way that arrhythmias cannot appear anymore. Hospitalizations of concerned patients decrease, quality of life and outcome rise. Consequently, VT ablation represents a growing field in electrophysiology, especially for patients with endocardial scars in ischemic heart disease after myocardial infarction. However, ablation of ventricular tachycardia remains one of the most challenging procedures in the electrophysiology lab. Precise scar definition and localization of abnormal potentials are critical for ablation success. The following manuscript describes the use of a multipolar mapping catheter and 3-dimensional (3D) mapping system to create a high density electro-anatomical map of the left ventricle including a precise scar representation as well as mapping of fractionated and late potentials in order to allow a highly accurate substrate modification.

With the following protocol, we provide an approach to Ventricular Tachycardia (VT) ablation using high density mapping with a multipolar catheter and 3D mapping system enhancing the success of the procedure.

Ablación de Taquicardia Ventricular isquémica usando un catéter Multipolar y el sistema de asignación 3-dimensional para la reconstrucción anatómica Electro alta densidad

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars ...

A Computerized Functional Skills Assessment and Training Program Targeting Technology Based Everyday Functional Skills

Today, many functional skills are technology-based, so development of a technology-based training program has broad importance. Here we present a computerized functional skills training program that was paired in half of the participants with a commercially available cognitive training (CCT) program.
Non-impaired older individuals (NC) aged 60+ (n=45) and similarly aged individuals with mild cognitive impairment (MCI; n=50) were randomized to receive 12 weeks of twice-weekly computerized functional skills training (CFST) or 12 weeks of twice-weekly sessions split between CCT and CFST. Skills trained were use of an ATM; internet banking; ticket kiosk; telephone and internet prescription refill; medication management; and internet shopping. As with previous functional capacity assessments, we focus on completion time for each simulation.
51 participants completed the training program, either by mastering all 6 tasks (34) or completing 12 weeks of training. 44 more participants completed 4 or more training sessions so they were also analyzed for improvement up to their last training session. Completion time for all 6 tests significantly improved from the baseline assessment to the final training session in both groups of participants (all p&#60;0.001 with an average improvement in task completion time of 45%). Further, there was no differential improvement in MCI and NC in the 6 tests from baseline to end of training (all t&#60;1.66, all p&#62;0.12). Finally, combined CCT plus CFST did not differ from CSFT alone on any of the percent-change score measures (all t&#60;1.64, all p&#62;0.11).
Both NC and MCI groups evidenced substantial improvements in performance. CCT supplementation led to similar functional gains with half as many training sessions. The NC participants proceeded through the training fairly rapidly even without CCT supplementation; MCI participants required more training but learned equivalently. These findings suggest that even in cases with memory impairments, functional skills can be efficiently learned with training.

This training protocol uses computerized training to teach technology-related everyday functional skills. These skills include financial skills, travel and transit, as well as medication management.

Un Programa Computarizado de Evaluación y Capacitación de Habilidades Funcionales Dirigido a Las Habilidades Funcionales Diarias Basadas en La Tecnología

Today, many functional skills are technology-based, so development of a technology-based training program has broad importance. Here we present a ...

A Reproducible Intensive Care Unit-Oriented Endotoxin Model in Rats

Sepsis and septic shock remain the leading cause of death in intensive care units. Despite significant improvements in sepsis management, mortality still ranges between 20 and 30%. Novel treatment approaches in order to reduce sepsis-related multiorgan failure and death are urgently needed. Robust animal models allow for one or multiple treatment approaches as well as for testing their effect on physiological and molecular parameters. In this article, a simple animal model is presented.
First, general anesthesia is induced in animals either with the use of volatile or by intraperitoneal anesthesia. After placement of an intravenous catheter (tail vein), tracheostomy, and insertion of an intraarterial catheter (tail artery), mechanical ventilation is started. Baseline values of mean arterial blood pressure, arterial blood oxygen saturation, and heart rate are recorded.
The injection of lipopolysaccharides (1 milligram/kilogram body weight) dissolved in phosphate-buffered saline induces a strong and reproducible inflammatory response via the toll-like receptor 4. Fluid corrections as well as the application of norepinephrine are performed based on well-established protocols.
The animal model presented in this article is easy to learn and strongly oriented towards clinical sepsis treatment in an intensive care unit with sedation, mechanical ventilation, continuous blood pressure monitoring, and repetitive blood sampling. Also, the model is reliable, allowing for reproducible data with a limited number of animals in accordance with the 3R (reduce, replace, refine) principles of animal research. While animal experiments in sepsis research cannot easily replaced, repetitive measurements allow for a reduction of animals and keeping septic animals anesthetized diminishes suffering.

Here, we present a reproducible intensive care unit-oriented endotoxin model in rats.

Un modelo de endotoxina reproducible orientado a la unidad de cuidados intensivos en ratas

Sepsis and septic shock remain the leading cause of death in intensive care units. Despite significant improvements in sepsis management, mortality still ...