Deseamos agradecer a un contrato (WS1763368) de Pfizer Pharmaceuticals para el apoyo en este trabajo. También nos gustaría dar las gracias a Arabia Saudí Ministerio de Educación Superior (Mohe) por su apoyo.

1. El ejercicio <ol><li> El músculo de interés para este experimento fue los flexores del codo (bíceps braquial). </li><li> La fuerza muscular se midió para cada participante sea capaz de dar a cada individuo una resistencia adecuada. Esto fue determinado por las pruebas a cada participante por su máxima resistencia (RM). </li><li> Para la prueba de la RM, se utilizó un dispositivo medidor de tensión conectado con una computadora a través de una Biopac (DA-100C) módulo de amplificación bioeléctrica (Biopac Systems, Goleta, CA) para medir la fuerza muscular. El módulo se conecta a un MP-100 analógica a la toma de muestras convertidor de señal digital a una frecuencia de 1.000 hertzios por segundo, y con una resolución de 24 bits (Fig. 10). </li><li> El aparato medidor de tensión se fija a un banco en un ángulo de 45 °. Los sujetos fueron instruidos para sentarse detrás del dispositivo y el resto de sus codos en el área de relleno, de modo que la fuerza de esfuerzo es a través de sus muñecas. Esta fue la mejor manera de asegurar que el tema no set contratar a cualquier otro músculo del bíceps (Fig. 11). </li><li> La fuerza se determinó en tres ocasiones con cada contracción siendo 3 segundos de duración con 45 segundos que separa las contracciones. El promedio de las tres mediciones fue la RM. </li><li> Después de la determinación de la RM para el músculo bíceps de cada participante, en la sesión prevista del ejercicio se llevó a cabo con el 35% de su RM. </li><li> Todos los sujetos fueron sometidos el mismo ejercicio con pesas adecuado ponderado para inducir el dolor muscular (DOMS). Esto se llevó a cabo al hacer 4 series de 25 repeticiones de curl de bíceps de concentración mientras se está sentado en una silla, y con los codos apoyados en los muslos (Fig. 12). </li><li> Cada sujeto recibió un periodo de 90 segundos de descanso entre cada serie. Los sujetos o hizo la serie completa de 25 repeticiones, o fueron instruidos para parar si no logra controlar constantemente el peso durante el ejercicio. </li></ol> 2. Infra-Red Camera Preparación y configuración <ol><li> La habitación en la que la proyección de imagen de infrarrojos se lleva a cabo se fijó en una temperatura constante para minimizar cualquier sesgo externo de las diferencias en la temperatura ambiente, lo que podría dar lugar a falsas lecturas térmicas. Con el propósito de este experimento nos dieron una habitación con temperatura controlada que se mantuvo a aproximadamente 23 ° C (+ / - 0.5 ° C). </li><li> La cámara se fijó en una distancia de 1 metro de distancia, y en un ángulo perpendicular a la piel que se mide (Fig. 9a) *. </li><li> Después de la distancia requerida se estableció, los sujetos se les aconseja quedarse quieto hasta que la imagen ha sido tomada. Esto no debe tomar más de un par de segundos, pero es muy importante para minimizar el movimiento para asegurar la exactitud de la imagen tomada. </li><li> Es preferible que la habitación tiene pintura de color más oscuro, en vez de los colores más claros, para minimizar cualquier interferencia infrarroja. </li><li> La iluminación es también fundamental cuando se trata de imágenes infrarrojas, becauso de fuente de luz que emiten ondas de infrarrojos como la iluminación fluorescente o de tungsteno puede dar lecturas falsas de alta. La mejor opción de iluminación sería una sala equipada con uniforme de luces LED, como las luces LED no producen ninguna interferencia infrarroja (Fig. 9b) *. </li></ol> * Una serie de pruebas se realizaron en nuestro laboratorio utilizando la cámara de infrarrojos FLIR 660 (Fig. 8), donde se compararon las imágenes de la piel en diferentes ángulos (0 (perpendicular), 15, 30, 45 y 60 grados), y a diferentes distancias (1, 2 y 5 metros) de la piel, para detectar con precisión la temperatura de la piel. Todas las imágenes fueron comparadas con termopares calibrados, y la mejor correlación entre las imágenes y las lecturas de los termopares se encontraba en un ángulo perpendicular y en una distancia de 1 metro de distancia de la piel (r = 0,93). Los diferentes ángulos y distancias causado una pérdida pixelización, y la disminución de la correlación general entre las imágenes y las lecturas del termopar. 3. Imagen de Acquirement <ol><li> Con el propósito de este experimento, la imagen del músculo ejercitado se tomó antes del ejercicio, ya las 24, y 48 horas post ejercicio. </li><li> El calor del cuerpo de otras fuentes que el objetivo podría afectar la imagen térmica y dar lecturas falsas. Por esta razón, nadie debe estar de pie al lado o detrás del objetivo. </li><li> En esta investigación, las imágenes de los dos ejerce el brazo y no ejerce-fueron tomadas para fines de comparación. Hemos ejercido uno de los brazos, como se mencionó anteriormente, y el otro brazo se utilizó como control (Fig. 4 y 5). </li><li> Números de la imagen de la cámara IR se registraron de inmediato en una hoja por separado, ya que podría ser difícil de identificar que la imagen pertenece a quién. </li></ol> 4. Procesamiento de imágenes y análisis <ol><li> Las imágenes adquiridas IR fueron procesados ​​mediante el &quot;ThermoVision ExaminIR&quot; Versión de software: 1.10.2. </li><li> Después de seleccionar la imagen requerida parar el análisis, cuatro regiones de interés fueron identificados en la imagen adquirida del brazo mediante las casillas de estadística en la interfaz del software (Fig. 6). </li><li> Cuando las regiones requiere de todo el brazo se han localizado, el programa muestra las medias y desviaciones estándar de las temperaturas para cada una de las regiones seleccionadas. A continuación, se puede comparar ni cruz cada región por separado o para obtener una temperatura media de todo el brazo (Fig. 7). </li></ol> 5. Escala Analógica Visual y Análisis de Sangre <ol><li> A escala visual analógica (EVA) se utilizó para evaluar el dolor subjetivo del brazo. La escala tuvo una de 10 cm (100mm) de largo, marcadas línea &quot;sin dolor&quot; en un extremo, y &quot;un dolor agudo&quot; en el extremo opuesto. A cada participante se dirige a hacer una marca a lo largo de la línea de 10 cm para indicar su respuesta al dolor. </li><li> El EVA se administraban a los sujetos antes del ejercicio, las 24 horas después del ejercicio, ya las 48 horas. </li><li> Sangre periférica se colseleccionado de los sujetos para medir los niveles de concentración de mioglobina en la sangre. </li><li> La sangre fue extraída de la vena antecubital temas antes del ejercicio, 30 minutos después de que el ejercicio había terminado, y en 48 horas. </li><li> La sangre se centrifuga a 4000 rpm durante 10 minutos para separar el suero de las células. Las muestras fueron almacenadas a -80 ° C hasta el análisis de la mioglobina se llevó a cabo. </li><li> La mioglobina se midió utilizando un TOSOH &quot;AIA-360&quot; Analizador Automatizado de inmunoensayo enzimático (TOSOH Corp., Tokio, Japón). Los kits de ensayo de la mioglobina (Myo 025297, ST AIA-PACK mioglobina) se utilizaron de acuerdo a las instrucciones del fabricante. </li></ol> 6. Resultados representante Los resultados de IR imágenes térmicas tomadas durante esta investigación están claramente representados en la figura 1. Imágenes tomadas en los 3 períodos de tiempo (antes del ejercicio, las 24 horas posteriores al ejercicio, y 48 horas post-ejercicio) para el ejercicio de las armas de los 41 sujetos, mostróun notable aumento de la temperatura en el día 2 (24 horas post-ejercicio) en comparación con antes del ejercicio temperaturas, y las temperaturas tomadas en 48 horas. Como se muestra en la figura 1, la temperatura promedio fue de 32,80 piel + / - 1,03 ° C para el día 1 (pre-ejercicio), y 33,96 + / - 1,46 ° C para el día 2 (24 horas post-ejercicio), y 32,82 + / - 1,29 para el día 3 (48 horas después del ejercicio). Esta diferencia en la temperatura de la piel a partir del día 1 al día 2 fue significativa (ANOVA p &lt;0,01). Sin embargo, para el grupo sin ejercicio, los cambios entre los 3 períodos de tiempo no eran evidentes. La figura 1 muestra que la temperatura promedio fue de 33,08 piel + / - 0,83 ° C para el día 1 (pre-ejercicio), y 32,79 + / - 1,42 ° C para el día 2 (24 horas post-ejercicio), y 33,17 + / - 0,95 para el día 3 (48 horas después del ejercicio). Esta diferencia en la temperatura de la piel durante los 3 días no fue significativa (ANOVA p = 0,38). Los resultados de las lecturas del dolor de la EVA se muestran en la figura 2. Como se observa en la figuragráfico 2, el dolor se había reportado un aumento dramático en los días 2 y 3. Los niveles de dolor del músculo ejerce aumentó de 3,6 + / - 6.1 en el día 1, a 36,3 + / - 22,8 el día 2, y 37.5 + / - 25,3 el día 3. Este incremento desde el día 1 fue significativa (ANOVA p &lt;0,01). Los resultados de los niveles de concentración de mioglobina se muestran en la figura 3. Como se ve en esta figura, casi no hubo cambios entre los 2 niveles de concentración de mioglobina en el día 1 (pre, y 30 minutos después del ejercicio). Sin embargo, el día 3, el aumento de la mioglobina era muy grande. Este aumento en el día 3 fue de aproximadamente 147 nanogramos por mililitro (ng / ml) de sangre, en comparación con los primeros 2 niveles de concentración en el día 1. Mioglobina fueron 30.12 + / - 7,66 ng / ml al inicio del estudio, 31,66 + / - 11,89 ng / mL 30 minutos después del ejercicio, y 178.96 + / - 249,51 ng / mL en el día 3. Este aumento en el día 3 fue altamente significativa (ANOVA p &lt;0,01). Un análisis de correlación se realizó entre la temperatura de la pielobtenidos de las imágenes de infrarrojos, y los niveles de dolor VAS. Se encontró que existe una correlación importante entre las lecturas de VAS en el día 2, y la medición de temperatura de la piel en el día 2. Esta correlación fue significativa (r = 0,312, p &lt;0,05). Sin embargo, no hubo una correlación evidente entre las lecturas de VAS y la temperatura de la piel en el día 3. Esta correlación fue significativa (r = 0,047, p = 0,77). <img src="/files/ftp_upload/3551/3551fig1.jpg" alt="Figura 1"/> Figura 1. Un gráfico representativo de las diferencias en la temperatura de la piel en los brazos ejercido (Diamantes), y sin ejercer las armas (cuadrados) de los 41 temas durante el periodo de 3 días. <img src="/files/ftp_upload/3551/3551fig2.jpg" alt="Figura 2"/> Figura 2. Un gráfico representativo de las diferencias en la percepción de dolor muscular medida con la VAS durante el período de tiempo de tres días para todos los 41 sujetos. Figura 3. Un gráfico representativo de las diferencias en las concentraciones de mioglobina de los 41 temas en los 3 períodos de tiempo. <img src="/files/ftp_upload/3551/3551fig4.jpg" alt="Figura 4"/> Figura 4. A) una imagen típica de infrarrojos del brazo ejerce un sujeto antes del ejercicio. B) una imagen de infrarrojos de los mismos temas del brazo 24 horas después del ejercicio. <img src="/files/ftp_upload/3551/3551fig5.jpg" alt="Figura 5"/> Figura 5. A) una imagen típica de un-IR ejerce el brazo de un sujeto antes del ejercicio. B) una imagen de infrarrojos de los mismos temas del brazo 24 horas después del ejercicio. <img src="/files/ftp_upload/3551/3551fig6.jpg" alt="Figura 6"/> Figura 6. Un ejemplo de las 4 regiones de interés para el análisis de la termiaal imagen del escudo. <img src="/files/ftp_upload/3551/3551fig7.jpg" alt="Figura 7"/> Figura 7. Interfaz de software para el &quot;ExaminIR ThermoVision&quot; que muestra las 4 casillas de interés en una imagen de infrarrojos de un brazo ejercido. También se muestran las interpretaciones estadísticas de cada caja. <img src="/files/ftp_upload/3551/3551fig8.jpg" alt="Figura 8"/> Figura 8. La cámara térmica de infrarrojos utilizada para esta investigación (FLIR 660). <img src="/files/ftp_upload/3551/3551fig9.jpg" alt="Figura 9"/> Figura 9. A) La configuración de la cámara IR 1 metro de distancia del brazo de los sujetos. B) Las luces LED que se utilizan en el laboratorio donde se tomaron las imágenes. <img src="/files/ftp_upload/3551/3551fig10.jpg" alt="Figura 10"/> Figura 10. A) Los módulos de Biopac utilizado para medir la fuerza muscular. B) El dispositivo medidor de tensión fija a un 45 º banco inclinado y conectado al sistema Biopac. <img src="/files/ftp_upload/3551/3551fig11.jpg" alt="Figura 11"/> Figura 11. Un tema típico de ejercer una fuerza sobre el aparato medidor de tensión. <img src="/files/ftp_upload/3551/3551fig12.jpg" alt="Figura 12"/> Figura 12. Un sujeto sometido el protocolo de ejercicio para inducir el dolor muscular.

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El propósito principal de esta investigación fue evaluar la utilidad de las imágenes de infrarrojos térmicos para detectar y medir el dolor muscular después del ejercicio vigoroso, y nuestros resultados sugieren que la imagen IR podría ser una técnica válida para la detección de DMAR, especialmente en las primeras 24 horas de ejercicio. Esto no es sorprendente, como Pennes 18, siempre un modelo muy detallado del flujo de calor desde el músculo a la piel en las extremidades. Este modelo predice que e...

<table border="1"><tbody><tr><td> Nombre del dispositivo </td><td> Empresa </td><td> Número de catálogo </td><td> Comentarios </td></tr><tr><td> Infra-Red cámara térmica </td><td> FLIR </td><td> FLIR SC660 </td><td></td></tr><tr><td> Térmica de infrarrojos Análisis de Software </td><td> ThermoVision ExaminIR </td><td> Software Version 1.10.2 </td><td></td></tr><tr><td> Módulo amplificador bioeléctrico </td><td> Biopac </td><td> DA100C </td><td> El DA100C proporciona la configuración de ganancia variable, y las referencias de tensión ajustable. </td></tr><tr><td> Análogo a Digital módulo convertidor </td><td> Biopac </td><td> MP100 </td><td></td></tr><tr><td> Analizador de Inmunoensayo enzimático automatizado </td><td> TOSOH </td><td> AFP -360 </td><td> Este dispositivo se utilizó para analizar las muestras de sangre, y obtener las lecturas de la mioglobina. </td></tr>
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the use of thermal infra-red imaging to detect delayed onset muscle soreness

Dolor muscular de aparición retardada (DOMS), también conocido como el daño muscular inducido por el ejercicio (IEGD), se experimenta comúnmente en individuos que han sido físicamente inactivos durante períodos prolongados de tiempo, y empezar una pelea inesperada del ejercicio 1.4, pero puede también ocurrir en atletas que se ejercitan más allá de sus límites normales de formación 5. Los síntomas asociados con este fenómeno doloroso puede variar desde dolor muscular leve, con dolor debilitante 1,3,5. La intensidad de estos síntomas y aumenta el malestar relacionados con las primeras 24 horas siguientes a la finalización del ejercicio, y los picos de entre 24 y 72 horas post ejercicio 1,3. Por esta razón, el DOMS es una de las formas recurrentes más comunes de lesiones deportivas que pueden afectar al rendimiento de un individuo, y se convierten en intimidar a los 1,4 muchos. Para las últimas 3 décadas, el fenómeno de preservativos ha ganado una cantidad considerablede interés entre los investigadores y especialistas en fisiología del ejercicio, los deportes, y los campos de la rehabilitación 6. Ha habido una serie de estudios publicados la investigación de este suceso doloroso en cuanto a sus mecanismos subyacentes, las intervenciones terapéuticas y estrategias preventivas 1-5,7-12. Sin embargo, es evidente por las publicaciones que el DOMS no es una patología fácil de cuantificar, ya que existe una cantidad amplia de la variabilidad entre las herramientas y métodos de medición utilizados para cuantificar esta condición 6. Es obvio que ningún acuerdo se ha hecho en una mejor medida de evaluación para DOMS, lo que hace difícil verificar si una intervención específica que realmente ayuda en la disminución de los síntomas asociados con este tipo de dolor o no. Por lo tanto, DMAR puede ser visto como algo ambiguo, ya que muchos estudios dependen de la medición de dolor mediante una escala visual analógica (EVA) 10,13-15, que es una cuestión subjetiva y no una medida objetiva. A pesar de que la agujaLas biopsias del músculo, y los niveles de proteínas myofibre podría ser considerado un estándar de oro para unos 6, grandes variaciones en algunas de estas proteínas de la sangre han sido documentados 6,16, además de los altos riesgos asociados a veces con técnicas invasivas. Por lo tanto, en la presente investigación, hemos probado una térmica de infrarrojos (IR) técnica de imagen de la piel sobre el músculo ejercitado para detectar el dolor muscular asociado. La termografía infrarroja se ha utilizado, y que han tenido éxito en la detección de diferentes tipos de enfermedades e infecciones desde 1950 17. Pero, sorprendentemente, casi nada se ha hecho sobre DOMS y los cambios en la temperatura de la piel. El objetivo principal de esta investigación fue examinar los cambios en el DOMS utilizando esta técnica segura y no invasiva.

Watch this Scientific Journal Video about The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness at JoVE.com

The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

El propósito de esta investigación fue evaluar si el uso de una cámara térmica de infrarrojos es una herramienta válida para detectar y cuantificar el dolor muscular después del ejercicio.

El uso de la térmica de infrarrojos de imagen para detectar el envaramiento

Delayed onset muscle soreness (DOMS), also known as exercise induced muscle damage (EIMD), is commonly experienced in individuals who have been physically ...

the-use-thermal-infra-red-imaging-to-detect-delayed-onset-muscle

Research

JoVE Journal

Medicine

19.1K vistas.  Loma Linda University. El propósito de esta investigación fue evaluar si el uso de una cámara térmica de infrarrojos es una herramienta válida para detectar y cuantificar el dolor muscular después del ejercicio.

Video: The Use of Thermal Infra-Red Imaging to Detect Delayed Onset Muscle Soreness

Thermal Ablation for the Treatment of Abdominal Tumors

Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ablation procedure, a thin applicator is guided into the target tumor under imaging guidance. Energy is then applied to the tissue until temperatures rise to cytotoxic levels (50-60 &deg;C). Various energy sources are available to heat biological tissues, including radiofrequency (RF) electrical current, microwaves, laser light and ultrasonic waves. Of these, RF and microwave ablation are most commonly used worldwide.
During RF ablation, alternating electrical current (~500 kHz) produces resistive heating around the interstitial electrode. Skin surface electrodes (ground pads) are used to complete the electrical circuit. RF ablation has been in use for nearly 20 years, with good results for local tumor control, extended survival and low complication rates1,2. Recent studies suggest RF ablation may be a first-line treatment option for small hepatocellular carcinoma and renal-cell carcinoma3-5. However, RF heating is hampered by local blood flow and high electrical impedance tissues (eg, lung, bone, desiccated or charred tissue)6,7. Microwaves may alleviate some of these problems by producing faster, volumetric heating8-10. To create larger or conformal ablations, multiple microwave antennas can be used simultaneously while RF electrodes require sequential operation, which limits their efficiency. Early experiences with microwave systems suggest efficacy and safety similar to, or better than RF devices11-13.
Alternatively, cryoablation freezes the target tissues to lethal levels (-20 to -40 &deg;C). Percutaneous cryoablation has been shown to be effective against RCC and many metastatic tumors, particularly colorectal cancer, in the liver14-16. Cryoablation may also be associated with less post-procedure pain and faster recovery for some indications17. Cryoablation is often contraindicated for primary liver cancer due to underlying coagulopathy and associated bleeding risks frequently seen in cirrhotic patients. In addition, sudden release of tumor cellular contents when the frozen tissue thaws can lead to a potentially serious condition known as cryoshock 16.
Thermal tumor ablation can be performed at open surgery, laparoscopy or using a percutaneous approach. When performed percutaneously, the ablation procedure relies on imaging for diagnosis, planning, applicator guidance, treatment monitoring and follow-up. Ultrasound is the most popular modality for guidance and treatment monitoring worldwide, but computed tomography (CT) and magnetic resonance imaging (MRI) are commonly used as well. Contrast-enhanced CT or MRI are typically employed for diagnosis and follow-up imaging.

A thermal tumor ablation procedure is described. The entire procedure is detailed, including pretreatment planning and imaging studies, anesthesia, adjuvant techniques to facilitate a percutaneous approach, imaging guidance of the ablation device to the tumor, thermal treatment, post-treatment care and follow-up.

La ablación térmica para el tratamiento de los tumores abdominales

Percutaneous thermal ablation is an emerging treatment option for many tumors of the abdomen not amenable to conventional treatments. During a thermal ...

Investigación

Medicina

Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment for melanoma remains surgical excision, therefore, the key to extended survival is early detection 2,3. Considering the large numbers of patients diagnosed every year and the limitations in accessing specialized care quickly, the development of objective in vivo diagnostic instruments to aid the diagnosis is essential. New techniques to detect skin cancer, especially non-invasive diagnostic tools, are being explored in numerous laboratories. Along with the surgical methods, techniques such as digital photography, dermoscopy, multispectral imaging systems (MelaFind), laser-based systems (confocal scanning laser microscopy, laser doppler perfusion imaging, optical coherence tomography), ultrasound, magnetic resonance imaging, are being tested. Each technique offers unique advantages and disadvantages, many of which pose a compromise between effectiveness and accuracy versus ease of use and cost considerations. Details about these techniques and comparisons are available in the literature 4.
Infrared (IR) imaging was shown to be a useful method to diagnose the signs of certain diseases by measuring the local skin temperature. There is a large body of evidence showing that disease or deviation from normal functioning are accompanied by changes of the temperature of the body, which again affect the temperature of the skin 5,6. Accurate data about the temperature of the human body and skin can provide a wealth of information on the processes responsible for heat generation and thermoregulation, in particular the deviation from normal conditions, often caused by disease. However, IR imaging has not been widely recognized in medicine due to the premature use of the technology 7,8 several decades ago, when temperature measurement accuracy and the spatial resolution were inadequate and sophisticated image processing tools were unavailable. This situation changed dramatically in the late 1990s-2000s. Advances in IR instrumentation, implementation of digital image processing algorithms and dynamic IR imaging, which enables scientists to analyze not only the spatial, but also the temporal thermal behavior of the skin 9, allowed breakthroughs in the field.
In our research, we explore the feasibility of IR imaging, combined with theoretical and experimental studies, as a cost effective, non-invasive, in vivo optical measurement technique for tumor detection, with emphasis on the screening and early detection of melanoma 10-13. In this study, we show data obtained in a patient study in which patients that possess a pigmented lesion with a clinical indication for biopsy are selected for imaging. We compared the difference in thermal responses between healthy and malignant tissue and compared our data with biopsy results. We concluded that the increased metabolic activity of the melanoma lesion can be detected by dynamic infrared imaging.

We demonstrated that malignant pigmented lesions with increased metabolic activity generate quantifiable amounts of heat and the measurement of the transient thermal response of the skin to a cooling excitation allows quantitative identification of melanoma and other skin cancers (vs. non-proliferative nevi) at an early stage of the disease.

Visualización y detección cuantitativa de cáncer de piel utilizando imágenes térmica dinámica

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment ...

Use of Animal Model of Sepsis to Evaluate Novel Herbal Therapies

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several techniques, including infusion of exogenous bacterial toxin (endotoxemia) or bacteria (bacteremia), as well as surgical perforation of the cecum by cecal ligation and puncture (CLP)1-3. CLP allows bacteria spillage and fecal contamination of the peritoneal cavity, mimicking the human clinical disease of perforated appendicitis or diverticulitis. The severity of sepsis, as reflected by the eventual mortality rates, can be controlled surgically by varying the size of the needle used for cecal puncture2. In animals, CLP induces similar, biphasic hemodynamic cardiovascular, metabolic, and immunological responses as observed during the clinical course of human sepsis3. Thus, the CLP model is considered as one of the most clinically relevant models for experimental sepsis1-3.
Various animal models have been used to elucidate the intricate mechanisms underlying the pathogenesis of experimental sepsis. The lethal consequence of sepsis is attributable partly to an excessive accumulation of early cytokines (such as TNF, IL-1 and IFN-&gamma;)4-6 and late proinflammatory mediators (e.g., HMGB1)7. Compared with early proinflammatory cytokines, late-acting mediators have a wider therapeutic window for clinical applications. For instance, delayed administration of HMGB1-neutralizing antibodies beginning 24 hours after CLP, still rescued mice from lethality8,9, establishing HMGB1 as a late mediator of lethal sepsis. The discovery of HMGB1 as a late-acting mediator has initiated a new field of investigation for the development of sepsis therapies using Traditional Chinese Herbal Medicine. In this paper, we describe a procedure of CLP-induced sepsis, and its usage in screening herbal medicine for HMGB1-targeting therapies.

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection, and can be simulated by a surgical technique termed cecal ligation and puncture (CLP). Here we describe a method to use CLP-induced animal model to screen medicinal herbs for therapeutic agents.

El uso de modelos animales de sepsis para evaluar nuevas terapias a base de plantas

Sepsis refers to a systemic inflammatory response syndrome resulting from a microbial infection. It has been routinely simulated in animals by several ...

The Use of Pharmacological-challenge fMRI in Pre-clinical Research: Application to the 5-HT System

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel underlying neurobiological mechanisms behind drug action and neurotransmitter-related disorders, such as depression and ADHD. Like most of the imaging methods (PET, SPECT, CT) it represents a progress in the investigation of brain disorders and the related function of neurotransmitter pathways in a non-invasive way with respect of the overall neuronal connectivity. Moreover it also provides the ideal tool for translation to clinical investigations. MRI, while still behind in molecular imaging strategies compared to PET and SPECT, has the great advantage to have a high spatial resolution and no need for the injection of a contrast-agent or radio-labeled molecules, thereby avoiding the repetitive exposure to ionizing radiations. Functional MRI (fMRI) is extensively used in research and clinical setting, where it is generally combined with a psycho-motor task. phMRI is an adaptation of fMRI enabling the investigation of a specific neurotransmitter system, such as serotonin (5-HT), under physiological or pathological conditions following activation via administration of a specific challenging drug.
The aim of the method described here is to assess brain 5-HT function in free-breathing animals. By challenging the 5-HT system while simultaneously acquiring functional MR images over time, the response of the brain to this challenge can be visualized. Several studies in animals have already demonstrated that drug-induced increases in extracellular levels of e.g. 5-HT (releasing agents, selective re-uptake blockers, etc) evoke region-specific changes in blood oxygenation level dependent (BOLD) MRI signals (signal due to a change of the oxygenated/deoxygenated hemoglobin levels occurring during brain activation through an increase of the blood supply to supply the oxygen and glucose to the demanding neurons) providing an index of neurotransmitter function. It has also been shown that these effects can be reversed by treatments that decrease 5-HT availability16,13,18,7. In adult rats, BOLD signal changes following acute SSRI administration have been described in several 5-HT related brain regions, i.e. cortical areas, hippocampus, hypothalamus and thalamus9,16,15. Stimulation of the 5-HT system and its response to this challenge can be thus used as a measure of its function in both animals and humans2,11.

The goal of this technique is to assess serotonin (5-HT) neurotransmitter function in the live and free-breathing animal with pharmacological magnetic resonance imaging (phMRI) and an intravenous challenge with a selective serotonin reuptake inhibitor (SSRI), fluoxetine.

El uso de la fMRI farmacológico-reto en la investigación preclínica: la aplicación al sistema de 5-HT

Pharmacological MRI (phMRI) is a new and promising method to study the effects of substances on brain function that can ultimately be used to unravel ...

Molecular Imaging to Target Transplanted Muscle Progenitor Cells

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle degeneration1, 2. In patients, the ability of resident muscle satellite cells (SCs) to regenerate damaged myofibers becomes increasingly inefficient4. Therefore, transplantation of muscle progenitor cells (MPCs)/myoblasts from healthy subjects is a promising therapeutic approach to DMD. A major limitation to the use of stem cell therapy, however, is a lack of reliable imaging technologies for long-term monitoring of implanted cells, and for evaluating its effectiveness. Here, we describe a non-invasive, real-time approach to evaluate the success of myoblast transplantation. This method takes advantage of a unified fusion reporter gene composed of genes (firefly luciferase [fluc], monomeric red fluorescent protein [mrfp] and sr39 thymidine kinase [sr39tk]) whose expression can be imaged with different imaging modalities9, 10. A variety of imaging modalities, including positron emission tomography (PET), single-photon emission computed tomography (SPECT), magnetic resonance imaging (MRI), optical imaging, and high frequency 3D-ultrasound are now available, each with unique advantages and limitations11. Bioluminescence imaging (BLI) studies, for example, have the advantage of being relatively low cost and high-throughput. It is for this reason that, in this study, we make use of the firefly luciferase (fluc) reporter gene sequence contained within the fusion gene and bioluminescence imaging (BLI) for the short-term localization of viable C2C12 myoblasts following implantation into a mouse model of DMD (muscular dystrophy on the X chromosome [mdx] mouse)12-14. Importantly, BLI provides us with a means to examine the kinetics of labeled MPCs post-implantation, and will be useful to track cells repeatedly over time and following migration. Our reporter gene approach further allows us to merge multiple imaging modalities in a single living subject; given the tomographic nature, fine spatial resolution and ability to scale up to larger animals and humans10,11, PET will form the basis of future work that we suggest may facilitate rapid translation of methods developed in cells to preclinical models and to clinical applications.

A non-invasive means to evaluate the success of myoblast transplantation is described. The method takes advantage of a unified fusion reporter gene composed of genes whose expression can be imaged with different imaging modalities. Here, we make use of a fluc reporter gene sequence to target cells via bioluminescence imaging.

Imagen Molecular a Target trasplantó células progenitoras musculares

Duchenne muscular dystrophy (DMD) is a severe genetic neuromuscular disorder that affects 1 in 3,500 boys, and is characterized by progressive muscle ...

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 ...

Murine Model for Non-invasive Imaging to Detect and Monitor Ovarian Cancer Recurrence

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of care consisting of surgical debulking and combination chemotherapy consisting of platinum and taxane compounds, almost 90% of patients recur within a few years. In these patients the development of chemoresistant disease limits the efficacy of currently available chemotherapy agents and therefore contributes to the high mortality. To discover novel therapy options that can target recurrent disease, appropriate animal models that closely mimic the clinical profile of patients with recurrent ovarian cancer are required. The challenge in monitoring intra-peritoneal (i.p.) disease limits the use of i.p. models and thus most xenografts are established subcutaneously. We have developed a sensitive optical imaging platform that allows the detection and anatomical location of i.p. tumor mass. The platform includes the use of optical reporters that extend from the visible light range to near infrared, which in combination with 2-dimensional X-ray co-registration can provide anatomical location of molecular signals. Detection is significantly improved by the use of a rotation system that drives the animal to multiple angular positions for 360 degree imaging, allowing the identification of tumors that are not visible in single orientation. This platform provides a unique model to non-invasively monitor tumor growth and evaluate the efficacy of new therapies for the prevention or treatment of recurrent ovarian cancer.

We describe a non-invasive animal imaging platform that allows the detection, quantification, and monitoring of ovarian cancer growth and recurrence. This intra-peritoneal xenograft model mimics the clinical profile of patients with ovarian cancer.

Modelo murino de imagen no invasivas para detectar y vigilar cáncer ovárico de Recurrencia

Epithelial ovarian cancer is the most lethal gynecologic malignancy in the United States. Although patients initially respond to the current standard of ...

Quantitative Magnetic Resonance Imaging of Skeletal Muscle Disease

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of living systems. Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using qMRI methods. The MRI acquisition protocol begins with localizer images (used to locate the position of the body and tissue of interest within the MRI system), quality control measurements of relevant magnetic field distributions, and structural imaging for general anatomical characterization. The qMRI portion of the protocol includes measurements of the longitudinal and transverse relaxation time constants (T1 and T2, respectively). Also acquired are diffusion-tensor MRI data, in which water diffusivity is measured and used to infer pathological processes such as edema. Quantitative magnetization transfer imaging is used to characterize the relative tissue content of macromolecular and free water protons. Lastly, fat-water MRI methods are used to characterize fibro-adipose tissue replacement of muscle. In addition to describing the data acquisition and analysis procedures, this paper also discusses the potential problems associated with these methods, the analysis and interpretation of the data, MRI safety, and strategies for artifact reduction and protocol optimization.

Neuromuscular diseases often exhibit a temporally varying, spatially heterogeneous, and multi-faceted pathology. The goal of this protocol is to characterize this pathology using non-invasive magnetic resonance imaging methods.

Imágenes por resonancia magnética cuantitativa de la enfermedad del músculo esquelético

Quantitative magnetic resonance imaging (qMRI) describes the development and use of MRI to quantify physical, chemical, and/or biological properties of ...

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation

Cardiovascular disease is the leading cause of morbidity and mortality in the world. Atherosclerotic plaques, consisting of lipid-laden macrophages and calcification, develop in the coronary arteries, aortic valve, aorta, and peripheral conduit arteries and are the hallmark of cardiovascular disease. In humans, imaging with computed tomography allows for the quantification of vascular calcification; the presence of vascular calcification is a strong predictor of future cardiovascular events. Development of novel therapies in cardiovascular disease relies critically on improving our understanding of the underlying molecular mechanisms of atherosclerosis. Advancing our knowledge of atherosclerotic mechanisms relies on murine and cell-based models. Here, a method for imaging aortic calcification and macrophage infiltration using two spectrally distinct near-infrared fluorescent imaging probes is detailed. Near-infrared fluorescent imaging allows for the ex vivo quantification of calcification and macrophage accumulation in the entire aorta and can be used to further our understanding of the mechanistic relationship between inflammation and calcification in atherosclerosis. Additionally, a method for isolating and culturing animal aortic vascular smooth muscle cells and a protocol for inducing calcification in cultured smooth muscle cells from either murine aortas or from human coronary arteries is described. This in vitro method of modeling vascular calcification can be used to identify and characterize the signaling pathways likely important for the development of vascular disease, in the hopes of discovering novel targets for therapy.

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

Calcificación de células del músculo liso y de imagen de la calcificación aórtica y la inflamación

Cardiovascular disease is the leading cause of morbidity and mortality in the world.  Atherosclerotic plaques, consisting of lipid-laden macrophages and ...

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

The tongue is a highly innervated and vascularized muscle hydrostat on the floor of the mouth of most vertebrates. Its primary functions include supporting mastication and deglutition, as well as taste-sensing and phonetics. Accordingly, the strength and volume of the tongue can impact the ability of vertebrates to accomplish basic activities such as feeding, communicating, and breathing. Human patients with sleep apnea have enlarged tongues, characterized by reduced muscle tone and increased intramuscular fat that can be visualized and quantified by magnetic resonance imaging (MRI). The abilities to measure force generation and viscoelastic properties of the tongue constitute important tools for obtaining functional information to correlate with imaging data. Here, we present techniques for measuring tongue force production in anesthetized Zucker rats via electrical stimulation of the hypoglossal nerves and for determining the viscoelastic properties of the tongue by applying passive Lissajous force/deformation curves.

In Vivo Evaluation of the Mechanical and Viscoelastic Properties of the Rat Tongue

We describe a surgical procedure in an anesthetized rat model for determining the muscle tone and viscoelastic properties of the tongue. The procedure involves specific stimulation of the hypoglossal nerves and application of passive Lissajous force/deformation curves to the muscle.