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1. Preparación de la placenta en el parto <ol><li> Etiqueta de los cordones umbilicales de los gemelos con un (a los primogénitos) o dos (para el segundo hijo) abrazaderas. </li><li> Inspeccione la superficie de la madre y el feto de la placenta de la integridad o la interrupción. </li><li> Registrar los siguientes datos: tipo de inserción del cordón umbilical (central, excéntrico, marginal o velamentosa), número de vasos sanguíneos en el cordón umbilical (normalmente una vena y dos arterias, a veces sólo una de las arterias) y la diferencia de color entre las dos acciones de la placenta. Una parte de las membranas dividen pueden ser enviados a patología para confirmar el tipo de corionicidad. </li><li> Entonces, la placenta se puede colocar en un recipiente de plástico y refrigeradas hasta el examen final (en el mejor plazo de una semana) y la inyección de colorantes. </li><li> La placenta no debe ser congelado o fijos (no utilice formol). </li></ol> 2. La cateterización de los vasos umbilicales <ol><li> Lave la placenta con agua tibia o solución salina. </li><li> Recorte las membranas periféricas, quitar la membrana entre dos divisorias y retire el amnios (para una mejor visualización de las anastomosis vasculares y una mejor calidad de las imágenes de la placenta). </li><li> Transecto cada cordón umbilical a una distancia de aproximadamente 5 cm de la inserción del cordón. </li><li> Apriete suavemente los coágulos de sangre de los vasos umbilicales y los vasos de la placenta. </li><li> La vena umbilical es generalmente fácil de identificar debido a su mayor diámetro, en comparación con el diámetro más pequeño de los dos arterias umbilicales. </li><li> Canular la vena umbilical con un catéter de tamaño adecuado. Evite pasajes falsos. </li><li> Canular una arteria umbilical con un catéter más pequeño. Use pinzas para ampliar la luz de la arteria umbilical. Evite pasajes falsos. Sólo una de las dos arterias umbilicales se debe cateterizar desde una anastomosis (de Hyrtl) conecta las 2 arterias cerca de la inserción del cordón umbilical. </li><li> Repita los pasos para que el cordón umbilical otros. </li><li> Colocación del catéter se puede facilitar de nuevo suave y hacia atrás de masaje de los vasos umbilicales. Cualquier tipo de catéter se puede usar para este procedimiento. Optamos por el uso (y el reciclaje) de los catéteres utilizados en nuestra sala de neonatología para la cateterización umbilical en recién nacidos. </li><li> Ate un pedazo de cinta adhesiva alrededor de los cables para evitar el reflujo del colorante durante la inyección de tinta. </li></ol> 3. Inyección con un tinte de color <ol><li> Conecte una jeringa de 20 ml llena de tintes de colores para cada catéter. </li><li> Un colorante de color viscoso se puede utilizar para visualizar la placenta angio-arquitectura. Use colores contrastantes para permitir una buena visualización de la anastomosis (los colores oscuros de las arterias, los colores brillantes de las venas). </li><li> Inyectar suavemente (con baja presión) el colorante en la vena, mientras que un ayudante empuja suavemente el medio de contraste para que el tinte de color para llenar todos los vasos de la placenta, también los más pequeños. </li><li> Preste especial atención a los pequeños vasos cerca del ecuador vascular (el ecuador vascular es el lugar donde las anastomosis de cualquiera de los mellizos se conectan unos con otros). </li><li> Repita los pasos anteriores para inyectar colorante en la arteria. De tener en cuenta: las arterias pueden ser más difíciles de inyectar y requieren más paciencia. </li><li> Repita los pasos para que el cordón umbilical otros. </li></ol> 4. Evaluación y documentación de la placenta después de la inyección colorante <ol><li> Examine cuidadosamente el ecuador vascular y registrar el número y tipo de anastomosis. </li><li> Coloque una cinta métrica en la placenta para medir los diámetros y las acciones de la placenta en la imagen digital. </li><li> Utilizar una cámara digital de alta resolución y tomar fotos de la placenta se inyecta. Asegúrese de que se tomen las imágenes perpendicular a la placenta. </li></ol> 5. Los resultados representativos: La angio-arquitectura de la placenta en los gemelos monocoriales, varía según el tipo de embarazo gemelar monocorial. Los estudios han demostrado que la inyección de anastomosis AA, AV y VV, respectivamente, están presentes en 80%, 95% y 20% de los embarazos monocoriales gemelos sin complicaciones 1 (Figura 1). Anastomosis AA se considera que protegen contra el desarrollo de TTTS y Grifos 6. Los estudios han demostrado que las inyecciones de anastomosis AA ocurren en sólo el 20% de TTTS placentas y el 10% de los grifos placentas 1,2,7 (Figura 2). En placentas TTTS tratarse con cirugía láser fetoscópica, las cicatrices causadas por la coagulación con láser de las anastomosis vasculares se pueden ver en la superficie de la placenta (Figura 3 y 4). Placentas GRIFOS se caracterizan por la presencia de sólo unos pocos minúscula anastomosis AV 2 (Figura 5). La participación de la placenta del destinatario TAPS es a menudo pletóricos, mientras que la cuota de la placenta de los donantes está pálido (Figura 6). En los embarazos gemelares monocoriales con la discordancia de peso al nacer, la restricción del crecimiento fetala menudo tiene una inserción del cordón velamentosa y un porcentaje mucho menor de la placenta (Figura 7). Anastomosis AA están a menudo presentes en las placentas de gemelos monocoriales, con la discordancia de peso al nacer 8. Monocoriónicos-monoamnióticos placentas tienen una característica en la angio-arquitectura con una distancia corta entre los dos inserciones del cordón umbilical (Figura 8). La incidencia de la anastomosis AA-en monoamnióticos monocoriales placentas es prácticamente del 100% e impide el desarrollo de TTTS en los gemelos monoamnióticos 9. Las cifras contenidas en este artículo muestran los hallazgos característicos de placentas monocoriales inyecta en nuestro centro con un tinte de color. Nosotros usamos los colores oscuros (azul o verde) para las arterias y los colores más claros (amarillo, rosa o naranja) para las venas. <img alt="Figura 1" src="/files/ftp_upload/3208/3208fig1.jpg" /> Figura 1. Placenta Monocoriónico de una normal, sin complicaciones de embarazo monocorial doble que muestra varias anastomosis AV de las arterias a las venas de color rosa verde (estrellas blancas), varias anastomosis VA de las arterias a las venas azul amarillo (estrellas verdes) y una gran anastomosis AA (identificados por la la mezcla de los tintes azules y verdes; estrella azul). <img alt="Figura 2" src="/files/ftp_upload/3208/3208fig2.jpg" /> Figura 2. Monocoriónico placenta en un embarazo TTTS tratados con amniorreducción serie que muestra la presencia de sólo AV (estrellas blancas) y VA (estrellas verdes) anastomosis sin una anastomosis AA. <img alt="Figura 3" src="/files/ftp_upload/3208/3208fig3.jpg" /> Figura 3. TTTS placenta después de la coagulación con láser fetoscópica de las anastomosis vasculares mediante la técnica de láser selectivo en el que se dejó una anastomosis residual pequeña sin darse cuenta de patentes (estrella blanca). Con la técnica de láser selectivo, las anastomosis vasculares se identificó por primera vez y, posteriormente, coagulada, uno por uno. <img alt="Figura 4" src="/files/ftp_upload/3208/3208fig4.jpg" /> Figura 4. TTTS placenta después de la coagulación con láser fetoscópica utilizando la técnica de Salomón en el que, después de la identificación y la coagulación de cada anastomosis individual, se coagula el ecuador vascular completa de un margen de la placenta con el otro. <img alt="Figura 5" src="/files/ftp_upload/3208/3208fig5.jpg" /> Figura 5. GRIFERIA placenta en el que sólo una anastomosis pocos minúscula VA (estrellas blancas) son visibles a lo largo del ecuador vascular. <img alt="Figura 6" src="/files/ftp_upload/3208/3208fig6.jpg" /> Figura 6. El lado materno de la placenta TAPS (se muestra en la Figura 5) muestra la diferencia de color entre las dos acciones de características de la placenta. <img alt="Figura 7" src="/files/ftp_upload/3208/3208fig7.jpg" /> Figura 7. Monocoriónico placenta de un embarazo gemelar, con la restricción selectiva del crecimiento intrauterino de uno de los gemelos. El feto tiene un crecimiento limitado inserción velamentosa del cordón y un porcentaje mucho menor de la placenta. La estrella blanca indica una anastomosis AA. <img alt="Figura 8" src="/files/ftp_upload/3208/3208fig8.jpg" /> Figura 8 monoamnióticos placenta:. Tenga en cuenta las diferentes AV (estrella verde), VA (estrella azul) y el 2 AA anastomosis (blanco estrellas), y la corta distancia entre las dos inserciones del cordón umbilical.

<ol>
	<li>Lopriore, E., Middeldorp, J. M., Sueters, M., Vandenbussche, F. P., Walther, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twin-to-twin+transfusion+syndrome:+from+placental+anastomoses+to+long-term+neurodevelopmental+outcome.">Twin-to-twin transfusion syndrome: from placental anastomoses to long-term neurodevelopmental outcome.</a> Curr. Pediatr. Rev. 1, 191-203 (2005).</li><li>Slaghekke, F., Kist, W. J., Oepkes, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Twin+anemia-polycythemia+sequence:+diagnostic+criteria,+classification,+perinatal+management+and+outcome.">Twin anemia-polycythemia sequence: diagnostic criteria, classification, perinatal management and outcome.</a> Fetal Diagn. Ther. 27, 181-190 (2010).</li><li>Lopriore, E., Middeldorp, J. M., Oepkes, D., Klumper, F. J., Walther, F. J., Vandenbussche, F. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Residual+anastomoses+after+fetoscopic+laser+surgery+in+twin-to-twin+transfusion+syndrome:+frequency,+associated+risks+and+outcome.">Residual anastomoses after fetoscopic laser surgery in twin-to-twin transfusion syndrome: frequency, associated risks and outcome.</a> Placenta. 28, 204-208 (2007).</li><li>Lopriore, E., Slaghekke, F., Middeldorp, J. M., Klumper, F. J., Oepkes, D., Vandenbussche, F. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Residual+anastomoses+in+twin-to-twin+transfusion+syndrome+treated+with+selective+fetoscopic+laser+surgery:+localization,+size,+and+consequences.">Residual anastomoses in twin-to-twin transfusion syndrome treated with selective fetoscopic laser surgery: localization, size, and consequences.</a> Am. J. Obstet. Gynecol. , (2009).</li><li>Lewi, L., Jani, J., Cannie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intertwin+anastomoses+in+monochorionic+placentas+after+fetoscopic+laser+coagulation+for+twin-to-twin+transfusion+syndrome:+is+there+more+than+meets+the+eye.">Intertwin anastomoses in monochorionic placentas after fetoscopic laser coagulation for twin-to-twin transfusion syndrome: is there more than meets the eye.</a> Am. J. Obstet. Gynecol. 194, 790-795 (2006).</li><li>Denbow, M. L., Cox, P., Taylor, M., Hammal, D. M., Fisk, N. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+angioarchitecture+in+monochorionic+twin+pregnancies:+relationship+to+fetal+growth,+fetofetal+transfusion+syndrome,+and+pregnancy+outcome.">Placental angioarchitecture in monochorionic twin pregnancies: relationship to fetal growth, fetofetal transfusion syndrome, and pregnancy outcome.</a> Am. J. Obstet. Gynecol. 182, 417-426 (2000).</li><li>van Meir, H., Slaghekke, F., Lopriore, E., van Wijngaarden, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arterio-arterial+anastomoses+do+not+prevent+the+development+of+twin+anemia-polycythemia+sequence.">Arterio-arterial anastomoses do not prevent the development of twin anemia-polycythemia sequence.</a> Placenta. 31, 163-165 (2010).</li><li>De Paepe, M. E., Shapiro, S., Young, L., Luks, F. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+characteristics+of+selective+birth+weight+discordance+in+diamniotic-monochorionic+twin+gestations.">Placental characteristics of selective birth weight discordance in diamniotic-monochorionic twin gestations.</a> Placenta. 31, 380-386 (2010).</li><li>Umur, A., van Gemert, M. J., Nikkels, P. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monoamniotic-versus+diamniotic-monochorionic+twin+placentas:+anastomoses+and+twin-twin+transfusion+syndrome.">Monoamniotic-versus diamniotic-monochorionic twin placentas: anastomoses and twin-twin transfusion syndrome.</a> Am. J. Obstet. Gynecol. 189, 1325-1329 (2003).</li><li>Senat, M. V., Deprest, J., Boulvain, M., Paupe, A., Winer, N., Ville, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endoscopic+laser+surgery+versus+serial+amnioreduction+for+severe+twin-to-twin+transfusion+syndrome.">Endoscopic laser surgery versus serial amnioreduction for severe twin-to-twin transfusion syndrome.</a> N. Engl. J. Med. 351, 136-144 (2004).</li><li>Lopriore, E., Sueters, M., Middeldorp, J. M., Vandenbussche, F. P., Walther, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Haemoglobin+differences+at+birth+in+monochorionic+twins+without+chronic+twin-to-twin+transfusion+syndrome.">Haemoglobin differences at birth in monochorionic twins without chronic twin-to-twin transfusion syndrome.</a> Prenat. Diagn. 25, 844-850 (2005).</li><li>Papathanasiou, D., Witlox, R., Oepkes, D., Walther, F. J., Bloemenkamp, K. W., Lopriore, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monochorionic+twins+with+ruptured+vasa+previa:+double+trouble!.">Monochorionic twins with ruptured vasa previa: double trouble!.</a> Fetal Diagn. Ther. 28, 48-50 (2010).</li><li>Weingertner, A. S., Kohler, A., Kohler, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+and+placental+characteristics+in+four+new+cases+of+twin+anemia-polycythemia+sequence.">Clinical and placental characteristics in four new cases of twin anemia-polycythemia sequence.</a> Ultrasound Obstet. Gynecol. 35, 490-494 (2010).</li><li>Lewi, L., Cannie, M., Blickstein, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+sharing,+birthweight+discordance,+and+vascular+anastomoses+in+monochorionic+diamniotic+twin+placentas.">Placental sharing, birthweight discordance, and vascular anastomoses in monochorionic diamniotic twin placentas.</a> Am. J. Obstet. Gynecol. 197, 587-588 (2007).</li><li>Lewi, L., Gucciardo, L., Huber, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+outcome+and+placental+characteristics+of+monochorionic+diamniotic+twin+pairs+with+early-+and+late-onset+discordant+growth.">Clinical outcome and placental characteristics of monochorionic diamniotic twin pairs with early- and late-onset discordant growth.</a> Am. J. Obstet. Gynecol. , (2008).</li><li>De Paepe, M. E., Friedman, R. M., Poch, M., Hansen, K., Carr, S. R., Luks, F. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Placental+findings+after+laser+ablation+of+communicating+vessels+in+twin-to-twin+transfusion+syndrome.">Placental findings after laser ablation of communicating vessels in twin-to-twin transfusion syndrome.</a> Pediatr. Dev. Pathol. 7, 159-165 (2004).</li><li>De Paepe, M. E., DeKoninck, P., Friedman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+distribution+patterns+in+monochorionic+twin+placentas.">Vascular distribution patterns in monochorionic twin placentas.</a> Placenta. 26, 471-475 (2005).</li><li>Nikkels, P. G., Hack, K. E., vanGemert, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pathology+of+twin+placentas+with+special+attention+to+monochorionic+twin+placentas.">Pathology of twin placentas with special attention to monochorionic twin placentas.</a> J. Clin. Pathol. 61, 1247-1253 (2008).</li><li>Quintero, R. A., Martinez, J. M., Lopez, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Individual+placental+territories+after+selective+laser+photocoagulation+of+communicating+vessels+in+twin-twin+transfusion+syndrome.">Individual placental territories after selective laser photocoagulation of communicating vessels in twin-twin transfusion syndrome.</a> Am. J. Obstet. Gynecol. 192, 1112-1118 (2005).</li><li>Quintero, R. A., Comas, C., Bornick, P. W., Allen, M. H., Kruger, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Selective+versus+non-selective+laser+photocoagulation+of+placental+vessels+in+twin-to-twin+transfusion+syndrome.">Selective versus non-selective laser photocoagulation of placental vessels in twin-to-twin transfusion syndrome.</a> Ultrasound Obstet. Gynecol. 16, 230-236 (2000).</li></ol>

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TTTS y la grifería son dos trastornos graves que pueden ocurrir en los gemelos monocoriales. Ambos trastornos se deben a la anastomosis AV causando un flujo sanguíneo desequilibrado intertwin. En las últimas 2 décadas, la cirugía láser fetoscópica estaban disponibles y ha demostrado ser el mejor método para el tratamiento de TTTS 10. El objetivo del tratamiento con láser fetoscópica es interrumpir la circulación entre los dos a través de la coagulación de las anastomosis vasculares en la superfic...

<table border="1"><tbody><tr><th> Nombre del reactivo </th><th> Empresa </th><th> Número de catálogo </th><th> Comentarios </th></tr><tr><td> Catéteres umbilicales (varias dimensiones, longitud 40 cm) </td><td> Vygon </td><td> 1,270.08 (ocho F) 1270.08 (5 F) 1270.04 (4 F) 1.270,03 (3,5 F) 1.270,02 (2,5 F) </td><td> Cualquier tipo de catéter se puede usar </td></tr><tr><td> 20 ml jeringas </td><td> BD Plastipak </td><td> 300613 </td><td></td></tr><tr><td> De tinta de color (colores diferentes) </td><td> Royal Talens Schoolverf </td><td> 36716010 (azul) 36715010 (verde) 36712350 (amarillo) 36713570 (rosa) </td><td> Un colorante de color viscoso se puede utilizar </td></tr></tbody></table>

accurate and simple evaluation of vascular anastomoses in monochorionic placenta using colored dye

La presencia de anastomosis vasculares placentarias es una condición sine qua non para el desarrollo del síndrome de transfusión gemelo a gemelo (TTTS) y la secuencia de dos policitemia anemia (TAPS) 1,2. Estudios de inyección de placentas gemelos han mostrado que las anastomosis son casi siempre presente en los gemelos monocoriales y extremadamente raro en dicoriónicos gemelos 1. Hay tres tipos de anastomosis se han documentado: desde la arteria a la arteria, la vena a vena y de la arteria a la vena. Arteriovenosa (AV) anastomosis son unidireccionales y se conocen como &quot;profundo&quot; anastomosis, ya que proceder a través de un cotiledón placentario compartida, mientras que arterio-arterial (AA) y veno-venosa (VV) anastomosis son bidireccionales y se conocen como &quot;superficial&quot; ya que se encuentran en la placa coriónica. Ambos TTTS y la grifería son causados ​​por el desequilibrio neto del flujo de sangre entre los gemelos debido a la anastomosis AV. Sangre de un gemelo (el donante) se bombea a través de una arteria en los cotiledones placentarios compartida y luego drena a través de una vena en la circulación de la otra gemela (el receptor). A menos que la sangre se bombea de nuevo desde el receptor al donante a través de direcciones opuestas anastomosis AV profunda o superficial a través de anastomosis, un desequilibrio del volumen de sangre se produce, poco a poco a la evolución de TTTS o grifos. La presencia de una anastomosis AA se ha demostrado que protege contra el desarrollo de TTTS y grifos para compensar el desequilibrio circulatorio causado por el 1,2 uni-direccional AV anastomosis. La inyección de placentas monocoriales, poco después del nacimiento es un medio útil para comprender la etiología de varios (hematológicos) complicaciones en los gemelos monocoriales, y es una prueba necesaria para alcanzar el diagnóstico de los TAPs 2. Además, la inyección de placentas TTTS tratarse con cirugía láser fetoscópica permite la identificación de posibles anastomosis residual 3-5. Esta información es de suma importancia para todos los perinatólogos involucrados en el manejo y cuidado de los gemelos monocoriónicos con TTTS o grifos. Varias técnicas de inyección de la placenta se están utilizando actualmente. Ofrecemos un protocolo simple para evaluar con precisión la presencia de anastomosis vasculares (residual) usando la inyección de colorante.

Watch this Scientific Journal Video about Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye at JoVE.com

Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

De gemelo a gemelo síndrome de transfusión de gemelo y la secuencia de la anemia policitemia dos problemas potencialmente devastadores en la medicina perinatal. Ambos trastornos se presentan sólo en los gemelos monocoriales, y el resultado del flujo de sangre a través de la placenta desequilibrada anastomosis vasculares. Ofrecemos un protocolo simple para evaluar con precisión la presencia de anastomosis vasculares mediante inyección de colorante de vasos placentarios después del nacimiento.

La evaluación precisa y simple de las anastomosis vasculares en la placenta monocoriónicos con tinte de color

The presence of placental vascular anastomoses is a conditio sine qua non for the development of twin-to-twin transfusion syndrome (TTTS) and twin anemia ...

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26.6K vistas.  Leiden University Medical Center. De gemelo a gemelo síndrome de transfusión de gemelo y la secuencia de la anemia policitemia dos problemas potencialmente devastadores en la medicina perinatal. Ambos trastornos se presentan sólo en los gemelos monocoriales, y el resultado del flujo de sangre a través de la placenta desequilibrada anastomosis vasculares. Ofrecemos un protocolo simple para evaluar con precisión la presencia de anastomosis vasculares mediante inyección de colorante de vasos placentari...

Video: Accurate and Simple Evaluation of Vascular Anastomoses in Monochorionic Placenta using Colored Dye

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&beta; using a Whole Blood Stimulation Assay

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues as well as altered cytokine homeostasis. The innate immune system plays a crucial role in recognizing pathogens and other endogenous danger stimuli. One of the major cytokines released by innate immune cells is Interleukin (IL)-1. Therefore, we utilize a whole blood stimulation assay in order to measure the secretion of inflammatory cytokines and specifically of the pro-inflammatory cytokine IL-1&beta; 1, 2, 3.
Patients with genetic dysfunctions of the innate immune system causing autoinflammatory syndromes show an exaggerated release of mature IL-1&beta; upon stimulation with LPS alone. In order to evaluate the innate immune component of patients who present with inflammatory-associated pathologies, we use a specific immunoassay to detect cellular immune responses to pathogen-associated molecular patterns (PAMPs), such as the gram-negative bacterial endotoxin, lipopolysaccharide (LPS). These PAMPs are recognized by pathogen recognition receptors (PRRs), which are found on the cells of the innate immune system 4, 5, 6, 7. A primary signal, LPS, in conjunction with a secondary signal, ATP, is necessary for the activation of the inflammasome, a multiprotein complex that processes pro-IL-1&beta; to its mature, bioactive form 4, 5, 6, 8, 9, 10.
The whole blood assay requires minimal sample manipulation to assess cytokine production when compared to other methods that require labor intensive isolation and culturing of specific cell populations. This method differs from other whole blood stimulation assays; rather than diluting samples with a ratio of RPMI media, we perform a white blood cell count directly from diluted whole blood and therefore, stimulate a known number of white blood cells in culture 2. The results of this particular whole blood assay demonstrate a novel technique useful in elucidating patient cohorts presenting with autoinflammatory pathophysiologies.

Accurate and Simple Measurement of the Pro-inflammatory Cytokine IL-1&amp;#946; using a Whole Blood Stimulation Assay

We describe a simple immunoassay to measure the production of pro-inflammatory cytokines, such as IL-1 beta production, in patients presenting with autoinflammatory phenotypes. By activating cells in whole blood cultures with pathogen-associated molecular patterns, specifically with lipopolysaccharide, cytokine secretion can be conveniently evaluated in whole blood supernatants.

La medición precisa y simple de la citoquina pro-inflamatoria IL-1β mediante un análisis de sangre entera estimulación

Inflammatory processes resulting from the secretion of soluble mediators by immune cells, lead to various manifestations in skin, joints and other tissues ...

Investigación

Medicina

Guide Wire Assisted Catheterization and Colored Dye Injection for Vascular Mapping of Monochorionic Twin Placentas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC twin pregnancies develop complications arising from the shared placenta and associated vascular connections1. Severe twin-to-twin syndrome (TTTS) is reported to account for approximately 20% of these complications2,3. Inter-twin vascular connections occur in almost all MC placentas and are related to the prognosis and outcome of these high-risk twin pregnancies. The number, size and type of connections have been implicated in the development of TTTS and other MC twin conditions. Three types of inter-twin vascular connections occur: 1) artery to vein connections (AVs) in which a branch artery carrying deoxygenated blood from one twin courses along the fetal surface of the placenta and dives into a placental cotyledon. Blood flows via a deep intraparenchymal capillary network into a draining vein that emerges at the fetal surface of the placenta and brings oxygenated blood toward the other twin. There is unidirectional flow from the twin supplying the afferent artery toward the twin receiving the efferent vein; 2) artery to artery connections (AAs) in which a branch artery from each twin meets directly on the superficial placental surface resulting in a vessel with pulsatile bidirectional flow, and 3) vein to vein connections (VVs) in which a branch vein from each twin meets directly on the superficial placental surface allowing low pressure bidirectional flow. In utero obstetric sonography with targeted Doppler interrogation has been used to identify the presence of AV and AA connections4. Prenatally detected AAs that have been confirmed by postnatal placental injection studies have been shown to be associated with an improved prognosis for both twins5. Furthermore, fetoscopic laser ablation of inter-twin vascular connections on the fetal surface of the shared placenta is now the preferred treatment for early, severe TTTS. 
Postnatal placental injection studies provide a valuable method to confirm the accuracy of prenatal Doppler ultrasound findings and the efficacy of fetal laser therapy6. Using colored dyes separately hand-injected into the arterial and venous circulations of each twin, the technique highlights and delineates AVs, AAs, and VVs. This definitive demonstration of MC placental vascular anatomy may then be correlated with Doppler ultrasound findings and neonatal outcome to enhance our understanding of the pathophysiology of MC twinning and its sequelae. Here we demonstrate our placental injection technique.

Vascular mapping of monochorionic (MC) twin placentas after birth provides a means for detailed demonstration of vascular connections between the twins&#x2019; circulations. 
Imbalance of these connections is thought to play a pivotal role in the development of complications of MC twinning including twin-to-twin transfusion syndrome.

Guía de alambre cateterismo asistida y de inyección de tinte de color de mapa vascular de Monocoriónico placentas Gemelas

Monochorionic (MC) twin pregnancies are associated with significantly higher morbidity and mortality rates than dichorionic twins. Approximately 50% of MC ...

A Simple Method of Mouse Lung Intubation

A simple procedure to intubate mice for pulmonary function measurements would have several advantages in longitudinal studies with limited numbers or expensive animal. One of the reasons that this is not done more routinely is that it is relatively difficult, despite there being several published studies that describe ways to achieve it. In this paper we demonstrate a procedure that eliminates one of the major hurdles associated with this intubation, that of visualizing the trachea during the entire time of intubation. The approach uses a 0.5 mm fiberoptic light source that serves as an introducer to direct the intubation cannula into the mouse trachea. We show that it is possible to use this procedure to measure lung mechanics in individual mice over a time course of at least several weeks. The technique can be set up with relatively little expense and expertise, and it can be routinely accomplished with relatively little training. This should make it possible for any laboratory to routinely carry out this intubation, thereby allowing longitudinal studies in individual mice, thereby minimizing the number of mice needed and increasing the statistical power by using each mouse as its own control.

This paper describes a striaghforward and efficient method of intubating mice for pulmonary function measurements or pulmonary instillation, that allows the mice to recover and be studied at later times. The procedure involves an inexpensive fiberoptic light source that directly illuminates the trachea.

Un sencillo método de intubación ratón de pulmón

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

Aspectos técnicos de la aortocava Fístula Ratón

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Fast and Accurate Exhaled Breath Ammonia Measurement

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that uses a quantum cascade based laser. The monitor is coupled to a sampler that measures mouth pressure and carbon dioxide. The system is temperature controlled and specifically designed to address the reactivity of this compound. The sampler provides immediate feedback to the subject and the technician on the quality of the breath effort. Together with the quick response time of the monitor, this system is capable of accurately measuring exhaled breath ammonia representative of deep lung systemic levels. 
Because the system is easy to use and produces real time results, it has enabled experiments to identify factors that influence measurements. For example, mouth rinse and oral pH reproducibly and significantly affect results and therefore must be controlled. Temperature and mode of breathing are other examples. As our understanding of these factors evolves, error is reduced, and clinical studies become more meaningful. This system is very reliable and individual measurements are inexpensive. 
The sampler is relatively inexpensive and quite portable, but the monitor is neither. This limits options for some clinical studies and provides rational for future innovations.

Ammonia is an important physiologic metabolite relevant to various disease and wellness states. It is also a difficult molecule to measure in breath, which demands particular precautions be taken to obtain accurate results. Not all factors influencing ammonia are known, but progress can be difficult without accounting for these factors.

Rápido y preciso exhalado Amoníaco Medición

This exhaled breath ammonia method uses a fast and highly sensitive spectroscopic method known as quartz enhanced photoacoustic spectroscopy (QEPAS) that ...

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or subvert anti-tumor immune responses in order to survive and flourish. Tumors take advantage of a number of different mechanisms of immune &#8220;escape,&#8221; including the recruitment of tolerogenic DC, immunosuppressive regulatory T cells (Tregs), and myeloid-derived suppressor cells (MDSC) that inhibit cytotoxic anti-tumor responses. Conversely, anti-tumor effector immune cells can slow the growth and expansion of malignancies: immunostimulatory dendritic cells, natural killer cells which harbor innate anti-tumor immunity, and cytotoxic T cells all can participate in tumor suppression. The balance between pro- and anti-tumor leukocytes ultimately determines the behavior and fate of transformed cells; a multitude of human clinical studies have borne this out. Thus, detailed analysis of leukocyte subsets within the tumor microenvironment has become increasingly important. Here, we describe a method for analyzing infiltrating leukocyte subsets present in the tumor microenvironment in a mouse tumor model. Mouse B16 melanoma tumor cells were inoculated subcutaneously in C57BL/6 mice. At a specified time, tumors and surrounding skin were resected en bloc and processed into single cell suspensions, which were then stained for multi-color flow cytometry. Using a variety of leukocyte subset markers, we were able to compare the relative percentages of infiltrating leukocyte subsets between control and chemerin-expressing tumors. Investigators may use such a tool to study the immune presence in the tumor microenvironment and when combined with traditional caliper size measurements of tumor growth, will potentially allow them to elucidate the impact of changes in immune composition on tumor growth. Such a technique can be applied to any tumor model in which the tumor and its microenvironment can be resected and processed.

This protocol describes a method for the detailed evaluation of leukocyte subsets within the tumor microenvironment in a mouse tumor model. Chemerin-expressing B16 melanoma cells were implanted subcutaneously into syngeneic mice. Cells from the tumor microenvironment were then stained and analyzed by flow cytometry, allowing for detailed leukocyte subset analyses.

Evaluación de tumor infiltrante de leucocitos subconjuntos en un modelo de tumor subcutáneo

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Using the Sleeve Technique in a Mouse Model of Aortic Transplantation - An Instructional Video

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the aorta in mice using the sleeve method was short and easy averaging 20 min, permitting studies of iso/allo grafts. The following article describes the aortic transplantation procedure used in our laboratory. The mice were anesthetized with a mixture of 1.5% volume isoflurane and 100% oxygen through a face mask. At this point, the segment of the aorta between the renal arteries and its bifurcation was separated from the vena cava, freely prepared and clampedat the proximal and distal segments with a single silk suture. Prior to the removal of the aorta, a saline solution containing heparin was injected into the inferior vena cava. Then the aorta was cut between the clamps and a saline heparin solution was used to flush the lumen. The sleeve technique with monofilament sutures was used in order to transplant the abdominal aorta in the orthotopic position.

We present an orthotopic aortic transplantation model using the sleeve technique in mice. It is a very rapid anastomosis method, which can be employed in studies of vascular disease.

Utilizando la técnica de la manga en un modelo murino de trasplante aórtico - un Video instructivo

Orthotopic aortic transplantation using the sleeve technique reduces injury to the aorta with failure rate of only 10-20%. The time to anastomose the ...

Model Surgical Training: Skills Acquisition in Fetoscopic Laser Photocoagulation of Monochorionic Diamniotic Twin Placenta Using Realistic Simulators

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome (TTTS), but is technically challenging and can lead to significant complications. Acquiring and maintaining the necessary surgical skills require consistent practice, a critical caseload, and time. Training on realistic surgical simulators can potentially shorten this steep learning curve and enables several proceduralists to acquire procedure-specific skills simultaneously. Here we describe realistic simulators designed to allow the user familiarity with the equipment and specific steps required in the surgical treatment of TTTS, including fetoscopic handling, approaches to anterior and posterior placenta, recognition of anastomoses, and efficient coagulation of vessels. We describe the skills that are especially important in conducting placental laser coagulation that the surgeon can practice on the model and apply in a clinical case. These models can be adapted easily depending on the availability of materials and require standard fetoscopy equipment. Such training systems are complementary to traditional surgical apprenticeships and can be useful aids for fetal medicine units that provide this clinical service.

Practicing the specific skills required for fetoscopic laser coagulation of monochorionic placental anastomoses on realistic models can aid less experienced surgeons in overcoming the steep learning curve associated with this procedure that is now regarded as the standard of care for twin-twin transfusion syndrome.

Entrenamiento quirúrgico modelo: Adquisición de habilidades en el photocoagulation del Laser Fetoscopic de Monochorionic Diamniotic Twin Placenta utilizando simuladores realistas

Fetoscopic laser coagulation of arterio-venous anastomoses (AVA) in a monochorionic placenta is the standard of care for twin-twin transfusion syndrome ...

Noninvasive Determination of Vortex Formation Time Using Transesophageal Echocardiography During Cardiac Surgery

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular (LV) filling compared with a continuous linear jet. Vortex ring development is most often quantified with vortex formation time (VFT), a dimensionless parameter based on fluid ejection from a rigid tube. Our group is interested in factors that affect LV filling efficiency during cardiac surgery. In this report, we describe how to use standard two-dimensional (2D) and Doppler transesophageal echocardiography (TEE) to noninvasively derive the variables needed to calculate VFT. We calculate atrial filling fraction (&#946;) from velocity-time integrals of trans-mitral early LV filling and atrial systole blood flow velocity waveforms measured in the mid-esophageal four-chamber TEE view. Stroke volume (SV) is calculated as the product of the diameter of the LV outflow track measured in the mid-esophageal long axis TEE view and the velocity-time integral of blood flow through the outflow track determined in the deep transgastric view using pulse-wave Doppler. Finally, mitral valve diameter (D) is determined as the average of major and minor axis lengths measured in orthogonal mid-esophageal bicommissural and long axis imaging planes, respectively. VFT is then calculated as 4 &#215; (1-&#946;) &#215; SV/(&#960;D3). We have used this technique to analyze VFT in several groups of patients with differing cardiac abnormalities. We discuss our application of this technique and its potential limitations and also review our results to date. Noninvasive measurement of VFT using TEE is straightforward in anesthetized patients undergoing cardiac surgery. The technique may allow cardiac anesthesiologists and surgeons to assess the impact of pathological conditions and surgical interventions on LV filling efficiency in real time.

We describe a protocol to measure vortex formation time, an index of left ventricular filling efficiency, using standard transesophageal echocardiography techniques in patients undergoing cardiac surgery. We apply this technique to analyze vortex formation time in several groups of patients with differing cardiac pathologies.

Determinación no invasiva del tiempo de formación de vórtice mediante ecocardiograma transesofágico durante la cirugía cardíaca

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular ...

Echocardiographic Evaluation of Atrial Communications before Transcatheter Closure

Transthoracic (TTE) and transesophageal echocardiography (TEE) is the standard imaging method for atrial septal defect (ASD) and patent foramen ovale (PFO) detection, for patient selection for transcatheter ASD/PFO closure,&#160;for intraoperative guidance and for long-term follow-up. The size, shape, location and the number of the atrial communications schould be determined. The accuracy of PFO detection can be improved by using agitated saline together with maneuvers to transiently increase the right atrial (RA) pressure. The appearance of microbubbles in the left atrium (LA) within 3 cardiac cycles after opacification of the RA is considered positive for the presence of an intracardiac shunt. Three dimensional TEE identifies further septal fenestrations and describes the dynamic morphology of ASD/PFO and atrial septal aneurysm. Follow-up evaluations with TTE is recommended at 1, 6, and 12 months after the procedure, with a subsequent evaluation every year. Previous studies showed an increased incidence of atrial arrhythmias early after device closure. Speckle tracking analysis may help to understand functional left atrial remodeling following percutaneous closure and its impact on atrial arrhythmias.

Transthoracic (TTE) and transesophageal (TEE) echocardiography represent the basic imaging tools for interatrial septum examination. Three dimensional (3D) TEE provides incremental information in the assessment of the interatrial septum. Further advanced echocardiography techniques using speckle tracking echocardiography are applied for sensitive volumetric and functional assessment of the heart chambers.&#160;

Evaluación ecocardiográfica de las comunicaciones auriculares antes del cierre transcatéter

Transthoracic (TTE) and transesophageal echocardiography (TEE) is the standard imaging method for atrial septal defect (ASD) and patent foramen ovale ...