Esta investigación fue apoyada por el Programa de Investigación Intramural del NIH, Instituto Nacional sobre el Envejecimiento y el Estudio Longitudinal de Envejecimiento de Baltimore. Una parte de ese apoyo fue a través de un contrato de I + D con el Instituto de Investigación MedStar. Protocolo estándar y la experiencia técnica fue proporcionada por AtCor.

1. Procedimiento <ol><li> Abra el software a través del acceso directo en el escritorio de Windows, haga doble clic en el icono. </li><li> Haga clic paciente para activar la pantalla del Paciente. </li><li> Para introducir un nuevo paciente en la base de datos, haga clic en el botón Nuevo en la zona de búsqueda paciente y, a continuación, haga clic en el botón Sí para confirmar que desea introducir un nuevo paciente. </li><li> Introduzca los datos del paciente, como apellido, nombre, fecha de nacimiento y sexo. Haga clic en el botón Guardar para avanzar a la siguiente etapa. </li><li> Haga clic en el botón de la VOP PWA o para llevar a cabo la medición. Si sólo uno de estos modos está disponible, no hay necesidad de hacer clic en el botón, basta con pasar a la pantalla del estudio. </li></ol> 2. Análisis de la onda de medición de la presión arterial central Usando pulsos (PWA) <ol><li> Haga clic en el botón de estudios para introducir los parámetros de estudio. Ingrese la presión braquial tomada de un esfigmomanómetro calibrado y cualquier otro detalle que desee incluir. </li><li> Click en el botón de datos de captura para proceder a una medición. Una los tres electrodos de ECG y conduce a la paciente en una configuración de plomo II modificado o el uso de las extremidades del paciente como se indica en los cables. </li><li> Palpar la arteria radial del paciente. Este se encuentra en el lateral de la muñeca, inmediatamente por encima de la articulación de la muñeca cerca de la base del pulgar. Presione suavemente la arteria radial contra el hueso subyacente con el medio y los dedos índices. </li><li> Coloque el tonómetro sobre el punto de pulso más fuerte. Presione suavemente el tonómetro abajo hasta que vea una forma de onda de presión constante que aparece en la pantalla de captura de datos. </li><li> Si Captura automática está habilitada (ver nota más adelante), el sistema guardará automáticamente la medición. Para guardar manualmente la medición para el análisis, pulse la barra espaciadora del teclado (o pise el pedal, si se utiliza). </li><li> Si se requiere una medición repetida con los mismos parámetros de estudio, haga clic en el botón Repetir para volver a la capturaPantalla de datos y repita la medición como se indica en los pasos 2.4 a 2.5 supra. </li><li> Para realizar una medición en un paciente nuevo, haga clic en el botón Paciente y vuelva al paso 1.3 anterior. </li></ol> 3. Rigidez arterial Medición Usando pulsos Velocidad de Onda (VOP) <ol><li> Haga clic en el botón de estudios para introducir los parámetros de estudio. </li><li> Ingrese la presión braquial tomada de un esfigmomanómetro calibrado Introduzca la distancia medida para el distal (supra-esternal muesca a pulso femoral) y proximal (supraesternal muesca a pulso de la carótida) sitios. Campos de la medicación, las notas y del operador puede introducir, si lo desea </li><li> Haga clic en el botón de captura para proceder a la medición. </li><li> Asegúrese de que la señal del ECG es de calidad aceptable. </li><li> Palpar el pulso de la arteria carótida. Este se encuentra entre la laringe y el músculo esternocleidomastoideo en el cuello. Se puede sentir empujando el músculo a un lado y presionando la arteria contra la laringe. </li><li&gt; Coloque el tonómetro sobre la arteria carótida y presione suavemente hacia abajo hasta que el tonómetro recibe una onda de presión constante que aparece en la pantalla de captura de datos. </li><li> Cuando usted tiene un mínimo de 13 segundos de formas de onda consistentes, haga clic en el botón OK o (prensa sobre el pedal, si se utiliza). </li><li> Palpar el pulso de la arteria femoral. Esta se encuentra sobre el muslo ventral a medio camino entre la sínfisis púbica y espina ilíaca anterosuperior. </li><li> Coloque el tonómetro en la arteria femoral y presione suavemente hacia abajo hasta que el tonómetro recibe una onda de presión constante que aparece en la pantalla de captura. Cuando usted tiene un mínimo de 13 segundos de formas de onda consistentes, haga clic en el botón OK (o pulse el interruptor de pie en el, si se utiliza). </li><li> En el informe (s) ahora puede ser revisada. Estas mediciones se llevan a cabo típicamente por triplicado. Si el usuario desea medir la VOP entre arterias carótidas y radiales, estos pasos sólo se muestran se repitieron excepto que en lugar de medir en la farteria emoral, el usuario coloque el tonómetro en la arteria radial. Para realizar una medición en un paciente nuevo, haga clic en el botón de Estudio y repita este procedimiento o utilizar la función de búsqueda para buscar un paciente existente. </li></ol> 4. Cerrar <ol><li> El software guarda automáticamente todos los informes. Para cerrar el programa, haga clic en Sistema en el menú principal y, a continuación, haga clic en Salir. Coloque el tonómetro en la bandeja del módulo para el almacenamiento. </li></ol>

<ol>
	<li>Najjar, S. S., Scuteri, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulse+wave+velocity+is+an+independent+predictor+of+the+longitudinal+increase+in+systolic+blood+pressure+and+of+incident+hypertension+in+the+Baltimore+Longitudinal+Study+of+Aging.">Pulse wave velocity is an independent predictor of the longitudinal increase in systolic blood pressure and of incident hypertension in the Baltimore Longitudinal Study of Aging.</a> J. Am. Coll. Cardiol. 51 (14), 1377-1383 (2008).</li><li>Doonan, R. J., Scheffler, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+arterial+stiffness+and+subendocardial+viability+ratio+in+young+healthy+light+smokers+after+acute+exercise.">Altered arterial stiffness and subendocardial viability ratio in young healthy light smokers after acute exercise.</a> PloS one. 6 (10), (2011).</li><li>Laurent, S., Cockcroft, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expert+consensus+document+on+arterial+stiffness:+methodological+issues+and+clinical+applications.">Expert consensus document on arterial stiffness: methodological issues and clinical applications.</a> Eur. Heart J. 27 (21), 2588-2605 (2006).</li><li>Mitchell, G. F., Hwang, S. -. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterial+stiffness+and+cardiovascular+events:+the+Framingham+Heart+Study.">Arterial stiffness and cardiovascular events: the Framingham Heart Study.</a> Circulation. 121 (4), 505-511 (2010).</li><li>Abbatecola, A. M., Chiodini, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulse+wave+velocity+is+associated+with+muscle+mass+decline:+Health+ABC+study.">Pulse wave velocity is associated with muscle mass decline: Health ABC study.</a> Age. , (2011).</li><li>Watson, N. L., Sutton-Tyrrell, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterial+stiffness+and+gait+speed+in+older+adults+with+and+without+peripheral+arterial+disease.">Arterial stiffness and gait speed in older adults with and without peripheral arterial disease.</a> Am. J. Hypertens. 24 (1), 90-95 (2011).</li><li>Mackenzie, I. S., Wilkinson, I. B., Cockcroft, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+arterial+stiffness+in+clinical+practice.">Assessment of arterial stiffness in clinical practice.</a> QJM. 95 (2), 67-74 (2002).</li><li>O'Rourke, M. F., Pauca, A., Jiang, X. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulse+wave+analysis.">Pulse wave analysis.</a> Br. J. Clin. Pharmacol. 51 (6), 507-522 (2001).</li><li>Stoner, L., Young, J. M., Fryer, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessments+of+arterial+stiffness+and+endothelial+function+using+pulse+wave+analysis.">Assessments of arterial stiffness and endothelial function using pulse wave analysis.</a> Int. J. Vasc. Med. 2012, 903107 (2012).</li><li>Farro, I., Bia, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulse+wave+velocity+as+marker+of+preclinical+arterial+disease:+reference+levels+in+a+uruguayan+population+considering+wave+detection+algorithms,+path+lengths,+aging,+and+blood+pressure.">Pulse wave velocity as marker of preclinical arterial disease: reference levels in a uruguayan population considering wave detection algorithms, path lengths, aging, and blood pressure.</a> Int. J. Hypertens. 2012, (2012).</li><li>Lakatta, E. G., Wang, M., Najjar, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterial+aging+and+subclinical+arterial+disease+are+fundamentally+intertwined+at+macroscopic+and+molecular+levels.">Arterial aging and subclinical arterial disease are fundamentally intertwined at macroscopic and molecular levels.</a> Med. Clin. N. Am. 93 (3), 583-604 (2009).</li><li>Ambrose, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Clinical Manual (CVMS) Rev. 2, 1-51 (2011).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reference+Values+for+Arterial+Stiffness'+Collaboration+Determinants+of+pulse+wave+velocity+in+healthy+people+and+in+the+presence+of+cardiovascular+risk+factors:+'establishing+normal+and+reference+values.">Reference Values for Arterial Stiffness' Collaboration Determinants of pulse wave velocity in healthy people and in the presence of cardiovascular risk factors: 'establishing normal and reference values.</a> Eur. Heart J. 31 (19), 2338-2350 (2010).</li><li>van Bortel, L. M., Laurent, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expert+consensus+document+on+the+measurement+of+aortic+stiffness+in+daily+practice+using+carotid-femoral+pulse+wave+velocity.">Expert consensus document on the measurement of aortic stiffness in daily practice using carotid-femoral pulse wave velocity.</a> J. Hypertens. 30 (3), 445-448 (2012).</li><li>Scuteri, A., Brancati, A., Gianni, W., Assisi, A., Volpe, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterial+stiffness+is+an+independent+risk+factor+for+cognitive+impairment+in+the+elderly:+a+pilot+study.">Arterial stiffness is an independent risk factor for cognitive impairment in the elderly: a pilot study.</a> J. Hypertens. 23 (6), 1211 (2005).</li><li>Khoshdel, A. R., Carney, S. L., Nair, B. R., Gillies, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Better+management+of+cardiovascular+diseases+by+pulse+wave+velocity:+combining+clinical+practice+with+clinical+research+using+evidence-based+medicine.">Better management of cardiovascular diseases by pulse wave velocity: combining clinical practice with clinical research using evidence-based medicine.</a> Clin. Med. Res. 5 (1), 45-52 (2007).</li></ol>

Los autores declaran que no tienen intereses financieros en competencia. Los autores recibieron ayuda con la descripción del protocolo y el rendimiento de estudio desde SphygmoCor que fabrica este producto descrito.

La medición de la velocidad de onda de pulso (VOP) es un método no invasivo para el análisis de la rigidez arterial central en una manera relativamente simple y reproducible 3. Para evaluar la VOP, la carótida (como un sustituto de la aorta descendente) y formas de onda de pulso femoral se adquirieron usando un tonómetro de alta fidelidad; el retardo de tiempo en el pulso de onda llegada a la referenciamiento entre los dos sitios se estima que un simultáneamente grabadas ondas EKG por los programas info...

A correction was made to <a href="http://www.jove.com/video/50817">Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging</a>. There was an error with the author, Majd AlGhatrif's, name. The author's name has been corrected to:
Majd AlGhatrif
instead of:
Majd al Ghatrif

A correction to the author list has been made for article <a href="http://www.jove.com/video/50817">Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging</a>.

Erratum: Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging

Pulso velocidad de la onda (VOP) es considerado como el estándar de oro en la evaluación de la rigidez arterial 2. Si bien no se utiliza clínicamente en los Estados Unidos, ha sido validado y ampliamente utilizado en el ámbito clínico en toda Europa 3. Los cambios en la VOP se han correlacionado con los factores de riesgo cardiovasculares tales como la hipertensión, la obesidad, intolerancia a la glucosa, y dislipidemia. Rigidez aórtica Superior se ha asociado con un mayor riesgo de sufrir un primer evento cardiovascular 4, la disminución de la masa muscular 5, y más lenta velocidad de la marcha en pacientes con enfermedad arterial periférica 6. Evaluación de la VOP se lleva a cabo mediante el análisis de la velocidad con la que un pulso de la sangre se desplaza desde la arteria carótida a la arteria femoral 7. Se trata de un método relativamente simple, con resultados altamente reproducibles. Los avances en la tecnología continúan para simplificar el procedimiento, la reducción de la dependencia del operador y disminuir la cantidad de tiempo requerido para laexamen. El sistema utilizado en nuestro laboratorio es producido por SphygmoCorand utiliza un electrocardiograma y un tonómetro de alta fidelidad para adquirir formas de onda de la carótida y pulsos femorales. Entonces, el programa analiza estas variables para determinar la velocidad por 1) estimar el tiempo que la onda de pulso se necesita para viajar entre la carótida y femoral sitios haciendo referencia a la temporización de la llegada de la onda de presión en cada sitio arterial en relación con las formas de onda electrocardiográficas 2) dividiendo la distancia medida entre los dos sitios de la hora prevista. Cuanto más cerca juntos los sitios de muestreo, la menos precisa la VOP, lo que limita el uso de este método para vasos más grandes, tales como las arterias femorales y carótidas. Además de determinar la VOP, el sistema puede calcular la presión en la aorta ascendente mediante la aplicación de una función de transferencia a la forma de onda producida por el pulso radial 8 que es la &quot;presión central&quot; y la presión que más afecta directamente el corazón en el tiempo de eyección. Esta información puede ayudar en la evaluación del efecto del sistema arterial en el ventrículo izquierdo de una manera fiable y no invasiva 9. Las técnicas alternativas utilizadas para el examen de la rigidez vascular, como la distensibilidad de la carótida a través de seguimiento de eco, requieren una gran cantidad de conocimientos técnicos y están más tiempo que la VOP. Seguimiento Echo también en general utiliza el análisis de imágenes de vídeo, por lo que la precisión de estas medidas se limitan 3. Otras técnicas para examinar los cambios vasculares incluyen la evaluación de la carótida-íntima media de espesor (IMT), que proporciona resultados más estrechamente relacionados con la progresión de la aterosclerosis, aunque las imágenes de cortes transversales y transversales obtenidos pueden proporcionar una aproximación burda de elastancia carótida. Este método, sin embargo, está plagado de algunas de las mismas limitaciones que los estudios de distensibilidad descritos anteriormente. Mientras los investigadores continúan buscando a los juegos de rigidez papel arteriales en diversos estados de enfermedad,Medición de la VOP probablemente seguirá proporcionando un criterio indirecto de valoración cuantitativa y reproducible.

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pulse wave velocity testing in the baltimore longitudinal study of aging

Velocidad de la onda de pulso carotídeo-femoral es considerado el estándar de oro para la medición de la rigidez arterial central obtenidos a través de métodos no invasivos 1. Los sujetos se colocaron en la posición supina y se dejan descansar en silencio durante al menos 10 min antes del comienzo del examen. Se selecciona el tamaño apropiado del manguito y se obtiene una presión arterial mediante un dispositivo oscilométrico. Una vez obtenida una presión arterial en reposo, ondas de presión se adquieren de las arterias carótidas comunes femorales y derecha derecha. El sistema calcula automáticamente el tiempo de tránsito del pulso entre estos dos sitios (usando la arteria carótida como un sustituto de la aorta descendente). Mediciones de la superficie del cuerpo se utilizan para determinar la distancia recorrida por la onda de pulso entre los dos sitios de muestreo. Esta distancia se divide por el tiempo de tránsito del pulso resultante en la velocidad de la onda de pulso. Las mediciones se realizaron por triplicado y el promedio se utiliza para anallisis.

<html><img alt="Tabla 1" fo:content-width="5in" fo:src="/files/ftp_upload/50817/50817table1highres.jpg" src="/files/ftp_upload/50817/50817table1.jpg" /> Tabla 1. Resultados PWV se producen en metros por segundo. En individuos sanos, estas velocidades pueden variar desde un promedio 6.2 m / s (en sujetos menores de 30 años) a 10,9 m / s (en sujetos de 70 años o más). Estos valores fueron establecidos en el artículo &quot;Factores determinantes de la velocida...

Watch this Scientific Journal Video about Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging at JoVE.com

Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging

Velocidad de la onda de pulso (VOP) de medición evalúa la rigidez arterial por un sistema basado en la tonometría que mide la velocidad con la que la onda de presión arterial viaja a lo largo del árbol arterial, típicamente aproximar el tiempo que tarda esta onda para viajar desde la aorta descendente (usando la carótida arteria como un sustituto) a la arteria femoral. -Carotídeo femoral VOP se incrementa de dos a tres veces durante toda la vida adulta.

Onda de pulso Pruebas de velocidad en el Baltimore Longitudinal Study of Aging

Carotid-femoral pulse wave velocity is considered the gold standard for measurements of central arterial stiffness obtained through noninvasive ...

pulse-wave-velocity-testing-baltimore-longitudinal-study

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Education

JoVE Core

JoVE Journal

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Medicine

Unit 1

Vital Signs: Pulse

SCIENTISTS IN ACTION

16.9K vistas.  National Institute of Aging.  Velocidad de la onda de pulso (VOP) de medición evalúa la rigidez arterial por un sistema basado en la tonometría que mide la velocidad con la que la onda de presión arterial viaja a lo largo del árbol arterial, típicamente aproximar el tiempo que tarda esta onda para viajar desde la aorta descendente (usando la carótida arteria como un sustituto) a la arteria femoral. -Carotídeo femoral VOP se incrementa de dos a tres veces durante toda la vida adulta. 

Video: Pulse Wave Velocity Testing in the Baltimore Longitudinal Study of Aging

Biomarkers in an Animal Model for Revealing Neural, Hematologic, and Behavioral Correlates of PTSD

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for objective diagnosis but also for the evaluation of therapeutic efficacy and resilience to trauma. Ongoing research is directed at identifying molecular biomarkers for PTSD, including traumatic stress induced proteins, transcriptomes, genomic variances and genetic modulators, using biologic samples from subjects' blood, saliva, urine, and postmortem brain tissues. However, the correlation of these biomarker molecules in peripheral or postmortem samples to altered brain functions associated with psychiatric symptoms in PTSD remains unresolved. Here, we present an animal model of PTSD in which both peripheral blood and central brain biomarkers, as well as behavioral phenotype, can be collected and measured, thus providing the needed correlation of the central biomarkers of PTSD, which are mechanistic and pathognomonic but cannot be collected from people, with the peripheral biomarkers and behavioral phenotypes, which can.
Our animal model of PTSD employs restraint and tail shocks repeated for three continuous days - the inescapable tail-shock model (ITS) in rats. This ITS model mimics the pathophysiology of PTSD 17, 7, 4, 10. We and others have verified that the ITS model induces behavioral and neurobiological alterations similar to those found in PTSD subjects 17, 7, 10, 9. Specifically, these stressed rats exhibit (1) a delayed and exaggerated startle response appearing several days after stressor cessation, which given the compressed time scale of the rat's life compared to a humans, corresponds to the one to three months delay of symptoms in PTSD patients (DSM-IV-TR PTSD Criterian D/E 13), (2) enhanced plasma corticosterone (CORT) for several days, indicating compromise of the hypothalamopituitary axis (HPA), and (3) retarded body weight gain after stressor cessation, indicating dysfunction of metabolic regulation.
The experimental paradigms employed for this model are: (1) a learned helplessness paradigm in the rat assayed by measurement of acoustic startle response (ASR) and a charting of body mass; (2) microdissection of the rat brain into regions and nuclei; (3) enzyme-linked immunosorbent assay (ELISA) for blood levels of CORT; (4) a gene expression microarray plus related bioinformatics tools 18. This microarray, dubbed rMNChip, focuses on mitochondrial and mitochondria-related nuclear genes in the rat so as to specifically address the neuronal bioenergetics hypothesized to be involved in PTSD.

We describe a rat model of post traumatic stress disorder (PTSD) that reveals the persistent alterations in neuroendocrine function and the delayed long-term, exaggerated fear response, characteristic of PTSD patients. The animal model and methods described here are useful for correlating biomarkers in brain nuclei, which are mechanistic but cannot be measured in patients, with biomarkers in peripheral white blood cells, which can.

Biomarcadores en un modelo animal para Revelar correlatos neurales, hematológicas y conductuales de estrés postraumático

Identification of biomarkers representing the evolution of the pathophysiology of Post Traumatic Stress Disorder (PTSD) is vitally important, not only for ...

Investigación

Medicina

Through the Looking Glass: Time-lapse Microscopy and Longitudinal Tracking of Single Cells to Study Anti-cancer Therapeutics

The response of single cells to anti-cancer drugs contributes significantly in determining the population response, and therefore is a major contributing factor in the overall outcome. Immunoblotting, flow cytometry and fixed cell experiments are often used to study how cells respond to anti-cancer drugs. These methods are important, but they have several shortcomings. Variability in drug responses between cancer and normal cells, and between cells of different cancer origin, and transient and rare responses are difficult to understand using population averaging assays and without being able to directly track and analyze them longitudinally. The microscope is particularly well suited to image live cells. Advancements in technology enable us to routinely image cells at a resolution that enables not only cell tracking, but also the observation of a variety of cellular responses. We describe an approach in detail that allows for the continuous time-lapse imaging of cells during the drug response for essentially as long as desired, typically up to 96 hr. Using variations of the approach, cells can be monitored for weeks. With the employment of genetically encoded fluorescent biosensors numerous processes, pathways and responses can be followed. We show examples that include tracking and quantification of cell growth and cell cycle progression, chromosome dynamics, DNA damage, and cell death. We also discuss variations of the technique and its flexibility, and highlight some common pitfalls.

Here, we describe a method of long-term time-lapse microscopy to longitudinally track single cells in response to anti-cancer therapeutics.

A través del espejo: Microscopía de lapso de tiempo y de seguimiento longitudinal de las células individuales para el Estudio de la terapéutica contra el cáncer

The response of single cells to anti-cancer drugs contributes significantly in determining the population response, and therefore is a major contributing ...

A Phenotyping Regimen for Genetically Modified Mice Used to Study Genes Implicated in Human Diseases of Aging

Age-related diseases are becoming increasingly prevalent and the burden continues to grow as our population ages. Effective treatments are necessary to lessen the impact of debilitating conditions but remain elusive in many cases. Only by understanding the causes and pathology of diseases associated with aging, can scientists begin to identify potential therapeutic targets and develop strategies for intervention. The most common age-related conditions are neurodegenerative disorders such as Parkinson's disease and blindness. Age-related macular degeneration (AMD) is the leading cause of blindness in the elderly. Genome wide association studies have previously identified loci that are associated with increased susceptibility to this disease and identified two regions of interest: complement factor H (CFH) and the 10q26 locus, where the age-related maculopathy susceptibility 2 (ARMS2) and high-temperature requirement factor A1 (HtrA1) genes are located. CFH acts as a negative regulator of the alternative pathway (AP) of the complement system while HtrA1 is an extracellular serine protease. ARMS2 is located upstream of HtrA1 in the primate genome, although the gene is absent in mice. To study the effects of these genes, humanized knock-in mouse lines of Cfh and ARMS2, knockouts of Cfh, HtrA1, HtrA2, HtrA3 and HtrA4 as well as a conditional neural deletion of HtrA2 were generated. Of all the genetically engineered mice produced only mice lacking HtrA2, either systemically or in neural tissues, displayed clear phenotypes. In order to examine these mice thoroughly and systematically, an initial phenotyping schedule was established, consisting of a series of tests related to two main diseases of interest: AMD and Parkinson's. Genetically modified mice can be subjected to appropriate experiments to identify phenotypes that may be related to the associated diseases in humans. A phenotyping regimen with a mitochondrial focus is presented here alongside representative results from the tests of interest.

A reverse-genetics approach to understanding gene families associated with human disease is presented, using mouse as a model system, and the subsequent mouse phenotyping schedule is described. Because mice defective in a gene of interest, HtrA2, manifested Parkinsonian symptoms, the phenotyping regimen is focused on identifying neurological defects.

Un régimen del fenotipo de ratones genéticamente modificados empleadas para el estudio genes implicados en enfermedades humanas de envejecimiento

Age-related diseases are becoming increasingly prevalent and the burden continues to grow as our population ages. Effective treatments are necessary to ...

Ultrasound-based Pulse Wave Velocity Evaluation in Mice

Arterial stiffness can be evaluated by calculating pulse wave velocity (PWV), i.e., the speed with which the pulse wave travels in a conduit vessel. This parameter is being increasingly investigated in small rodent models in which it is used for assessing alterations in vascular function related to particular genotypes/treatments or for characterizing cardiovascular disease progression. This protocol describes an image processing algorithm which leads to non-invasive arterial PWV measurement in mice using ultrasound (US) images only. The proposed technique has been used to assess abdominal aorta PWV in mice and evaluate its age-associated changes.
Abdominal aorta US scans are obtained from mice under gaseous anesthesia using a specific US device equipped with high-frequency US probes. B-mode and Pulse-Wave Doppler (PW-Doppler) images are analyzed in order to obtain diameter and mean velocity instantaneous values, respectively. For this purpose, edge detection and contour tracking techniques are employed. The single-beat mean diameter and velocity waveforms are time aligned and combined in order to achieve the diameter-velocity (lnD-V) loop. PWV values are obtained from the slope of the linear part of the loop, which corresponds to the early systolic phase.
With the present approach, anatomical and functional information about the mouse abdominal aorta can be non-invasively achieved. Requiring the processing of US images only, it may represent a useful tool for the non-invasive characterization of different arterial sites in the mouse in terms of elastic properties. The application of the present technique can be easily extended to other vascular districts, such as the carotid artery, thus providing the possibility to obtain a multi-site arterial stiffness assessment.

Arterial stiffness represents a key factor in cardiovascular disease and pulse wave velocity (PWV) can be considered as a surrogate index for arterial stiffness. This protocol describes an image processing algorithm for calculating PWV in mice based on ultrasound image processing that is applicable at different arterial sites.

El ultrasonido basados ​​en la onda de pulso Evaluación de velocidad en ratones

Arterial stiffness can be evaluated by calculating pulse wave velocity (PWV), i.e., the speed with which the pulse wave travels in a conduit vessel. This ...

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy (rNAION)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their lifetime. NAION results in optic nerve edema, selective loss of the retinal ganglion cell neurons (RGCs) and atrophy of the optic nerve. A rodent model of NAION that expresses most NAION features and sequelae has been developed, which is applicable to both rats and mice. This model utilizes a focal laser application of 532 nm wavelength to illuminate a photoactive dye, Rose Bengal (RB), to cause capillary damage and leakage at the targeted anterior optic nerve (the laminar region). After rNAION induction, there is an early optic nerve ischemia, optic nerve edema, and intraneural inflammation, followed by selective RGC and axonal loss. Since the optic nerve is a CNS white matter tract, the rNAION model is applicable to mechanistic studies of selective white matter ischemia, as well as neuroprotective analyses and short and long-term mechanisms of glial and neuronal response to ischemia.

The Rodent Model of Nonarteritic Anterior Ischemic Optic Neuropathy rNAION

The following report describes how to replicate the rodent model of nonarteritic anterior ischemic optic neuropathy (rNAION), using the appropriate dye, contact lens and laser parameters. We also reveal the appropriate steps for evaluating the rNAION lesion in vivo.

El modelo de roedor de la anterior no arterítica neuropatía óptica isquémica (rNAION)

Nonarteritic anterior ischemic optic neuropathy (NAION) is a focal ischemic lesion of the optic nerve that affects 1/700 individuals throughout their ...

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular plasticity or damage occurring overtime and in relation to neuronal activity or blood flow. In vivo two-photon microscopy allows the study of the structural and functional plasticity of large cellular units in the living brain. In particular, the thinned-skull window preparation allows the visualization of cortical regions of interest (ROI) without inducing significant brain inflammation. Repetitive imaging sessions of cortical ROI are feasible, providing the characterization of disease hallmarks over time during the progression of numerous CNS diseases. This technique accessing the pial structures within 250 &#956;m of the brain relies on the detection of fluorescent probes encoded by genetic cellular markers and/or vital dyes. The latter (e.g., fluorescent dextrans) are used to map the luminal compartment of cerebrovascular structures. Germane to the protocol described herein is the use of an in vivo marker of amyloid deposits, Methoxy-O4, to assess Alzheimer&#39;s disease (AD) progression. We also describe the post-acquisition image processing used to track vascular changes and amyloid depositions. While focusing presently on a model of AD, the described protocol is relevant to other CNS disorders where pathological cerebrovascular changes occur.

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

This manuscript describes a procedure to track the remodeling of the cerebrovasculature during amyloid plaque accumulation in vivo using longitudinal two-photon microscopy. A thinned-skull preparation enables the visualization of fluorescent dyes to assess the progression of cerebrovascular damage in a mouse model of Alzheimer&#39;s disease.

Longitudinal<em&gt; En Vivo</em&gt; Obtención de imágenes de la vasculatura cerebral: Importancia de las enfermedades del sistema nervioso central

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular ...

Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is often capped with biocompatible materials in order to expedite pulpal wound healing. The ultimate goal is to regenerate reparative dentin, a physical barrier that functions as a "biological seal" and protects the underlying pulp tissue. Although this direct pulp-capping procedure has long been used in dentistry, the underlying molecular mechanism of pulpal wound healing and reparative dentin formation is still poorly understood. To induce reparative dentin, pulp capping has been performed experimentally in large animals, but less so in mice, presumably due to their small sizes and the ensuing technical difficulties. Here, we present a detailed, step-by-step method of performing a pulp-capping procedure in mice, including the preparation of a Class-I-like cavity, the placement of pulp-capping materials, and the restoration procedure using dental composite. Our pulp-capping mouse model will be instrumental in investigating the fundamental molecular mechanisms of pulpal wound healing in the context of reparative dentin in vivo by enabling the use of transgenic or knockout mice that are widely available in the research community.

We describe a step-by-step method of performing direct pulp capping on mice teeth for the evaluation of pulpal wound healing and reparative dentin formation in vivo.

Desarrollo de un Modelo de recubrimiento pulpar directo para la Evaluación de la Pulpa Curación de Heridas y formación de dentina reparadora en ratones

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is ...

Using Retinal Imaging to Study Dementia

The retina offers a unique &#8220;window&#8221; to study pathophysiological processes of dementia in the brain, as it is an extension of the central nervous system (CNS) and shares prominent similarities with the brain in terms of embryological origin, anatomical features and physiological properties.&#160; The vascular and neuronal structure in the retina can now be visualized easily and non-invasively using retinal imaging techniques, including fundus photography and optical coherence tomography (OCT), and quantified semi-automatically using computer-assisted analysis programs. Studying the associations between vascular and neuronal changes in the retina and dementia could improve our understanding of dementia and, potentially, aid in diagnosis and risk assessment.&#160; This protocol aims to describe a method of quantifying and analyzing retinal vasculature and neuronal structure, which are potentially associated with dementia. This protocol also provides examples of retinal changes in subjects with dementia, and discusses technical issues and current limitations of retinal imaging.

The retina shares prominent similarities with the brain and thus represents a unique window to study vasculature and neuronal structure in the brain non-invasively. This protocol describes a method to study dementia using retinal imaging techniques. This method can potentially aid in diagnosis and risk assessment of dementia.

Usando proyección de imagen retiniana para el estudio de la demencia

The retina offers a unique &#8220;window&#8221; to study pathophysiological processes of dementia in the brain, as it is an extension of the central ...

Measuring the Carotid to Femoral Pulse Wave Velocity (Cf-PWV) to Evaluate Arterial Stiffness

For the elderly, arterial stiffening is a good marker for aging evaluation and it is recommended that the arterial stiffness be determined noninvasively by the measurement of carotid to femoral pulse wave velocity (cf-PWV) (Class I; Level of Evidence A). In literature, numerous community-based or disease-specific studies have reported that higher cf-PWV is associated with increased cardiovascular risk. Here, we discuss strategies to evaluate arterial stiffness with cf-PWV. Following the well-defined steps detailed here, e.g., proper position operator, distance measurement, and tonometer position, we will obtain a standard cf-PWV value to evaluate arterial stiffness. In this paper, a detailed stepwise method to record a good quality PWV and pulse wave analysis (PWA) using a non-invasive tonometry-based device will be discussed.

Measuring the Carotid to Femoral Pulse Wave Velocity Cf-PWV to Evaluate Arterial Stiffness

This protocol describes a method to standardize the measurements of carotid-femoral pulse wave velocity to evaluate arterial stiffness.

Medición de la carótida a la velocidad de la onda de pulso Femoral (Cf-PWV) para evaluar la rigidez Arterial

For the elderly, arterial stiffening is a good marker for aging evaluation and it is recommended that the arterial stiffness be determined noninvasively ...

Integration of Brain Tissue Saturation Monitoring in Cardiopulmonary Exercise Testing in Patients with Heart Failure

Cerebral hypo-oxygenation during rest or exercise negatively impacts the exercise capacity of patients with heart failure with reduced ejection fraction (HF). However, in clinical cardiopulmonary exercise testing (CPET), cerebral hemodynamics is not assessed. NIRS is used to measure cerebral tissue oxygen saturation (SctO2) in the frontal lobe. This method is reliable and valid and has been utilized in several studies. SctO2 is lower during both rest and peak exercise in patients with HF than in healthy controls (66.3 &#177; 13.3% and 63.4 &#177; 13.8% vs. 73.1 &#177; 2.8% and 72 &#177; 3.2%). SctO2 at rest is significantly linearly correlated with peak VO2 (r = 0.602), oxygen uptake efficiency slope (r = 0.501), and brain natriuretic peptide (r = -0.492), all of which are recognized prognostic and disease severity markers, indicating its potential prognostic value. SctO2 is determined mainly by end-tidal CO2 pressure, mean arterial pressure, and hemoglobin in the HF population. This article demonstrates a protocol that integrates SctO2 using NIRS into incremental CPET on a calibrated bicycle ergometer.

This protocol integrated near-infrared spectroscopy into conventional cardiopulmonary exercise testing to identify the involvement of the cerebral hemodynamic response in exercise intolerance in patients with heart failure.

Integración de la monitorización de la saturación del tejido cerebral en las pruebas de ejercicio cardiopulmonar en pacientes con insuficiencia cardíaca

Cerebral hypo-oxygenation during rest or exercise negatively impacts the exercise capacity of patients with heart failure with reduced ejection fraction ...