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4. Diagnóstico molecular de SARS-CoV-2

Author

Claudia Andrea Cruz B, Jeannette Navarrete O, and Gladys Pinilla

Subjects
0301 basic medicine, diagnosis, business.industry, 030106 microbiology, coronavirus, SARS virus, diagnóstico, 03 medical and health sciences, 0302 clinical medicine, virus SARS, Medicine, 030212 general & internal medicine, RT qPCR, business
Abstract

COVID-19 diagnosis is based on both clinical aspects and screening tests. However, clinical symptoms and signs in infected patients are highly atypical; hence, molecular tests are essential for diagnosis. RT-qPCR is carried out at BSL II level laboratories; the main molecular targets for viral detection are E gene (envelope), and RdRP gene (RNA-dependent RNA polymerase). False negatives in this diagnosis are due to sample quality and quantity, transport conditions, storage and handling before and after extraction (RNA is heat-labile and RNases are abundant); infection phase; virus mutations and presence of CRP inhibitors. Taking into account analytical sensitivity of RT-qPCR (5.2 copies of RNA / reaction) and the fact that once RNA it is extracted, it progressively degrades and affects test diagnostic sensitivity, a new sample -specifically taken from the lower respiratory tract in order to increase viral load- is recommended in the abovementioned cases. Timely diagnosis allows optimizing management (isolation and treatment), patient monitoring, implementing prevention and control measures as well as epidemiological surveillance of the disease El diagnóstico de COVID-19 se basa tanto en aspectos clínicos como en pruebas de detección, pero los síntomas y signos clínicos de los pacientes infectados son altamente atípicos y, por lo tanto, las pruebas moleculares son indispensables para su diagnóstico. La reacción en cadena de la polimerasa con transcriptasa inversa (RT-qPCR) se lleva a cabo en laboratorios nivel BSL II; asimismo, los principales blancos moleculares para la detección viral son el gen E (envoltura), y el gen RdRP (ARN polimerasa dependiente de ARN). Los falsos negativos en este diagnóstico se deben a la calidad y cantidad de la muestra, condiciones de transporte, almacenamiento, y manejo de estas antes y después de la extracción (el ARN es termolábil y abundantes las RNasas), fase de la infección, mutaciones del virus y presencia de inhibidores de la PCR. En estos casos, se recomienda una nueva toma de muestra, especialmente de vías respiratorias bajas, para aumentar la carga viral. Se debe tener en cuenta la sensibilidad analítica de la RT-qPCR (5,2 copias de ARN/reacción) y que, una vez el RNA se extrae, se va degradando progresivamente afectando la sensibilidad diagnóstica de la prueba. Un diagnóstico oportuno permite optimizar el manejo (aislamiento y tratamiento) y monitorización de los pacientes, así como la aplicación de medidas de prevención y control de la expansión, y la vigilancia epidemiológica de la enfermedad.

Published
2020
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5. Propuesta de diseño de una red inalámbrica de sensores y actuadores para riego, con tecnología de Internet de las Cosas

Author

Tovar Soto, Jhonatan Paolo, García Navarrete, O. L., Gutiérrez Martínez, Juan Carlos, Tovar Soto, Jhonatan Paolo, García Navarrete, O. L., and Gutiérrez Martínez, Juan Carlos

Abstract

Objetivo: Este trabajo presenta una selección detallada de tecnologías IoT para el diseño de una infraestructura de monitoreo de humedad ambiental para el control del riego de cultivos. Metodología: El diseño del prototipo se apoyó en estudios previos realizados por los autores y en pruebas realizadas en 2019 en la Finca el Porvenir en Cundinamarca, Colombia, con el fin de determinar la mejor opción para implementar en campo mediante el uso del software Python, y elementos como la Raspberry Pi y la placa de desarrollo TTGO. Resultados: iPara determinar la funcionalidad del software y el hardware del sistema, se realizaron diferentes pruebas estáticas del prototipo a través del monitoreo de humedad y temperatura ambiente por medio del sensor DHT22, cuya información es enviada desde un nodo transmisor a un nodo receptor en un concentrador para, posteriormente, guardar la información en una base de datos y, finalmente, obtener las gráficas del comportamiento de la humedad relativa, la temperatura, y el accionamiento del elemento de control (válvula solenoide). Conclusiones: Con esta información es posible demostrar la efectividad del prototipo, la posibilidad de conexión remota con el protocolo LoRa y el manejo y gestión de la información de diferentes zonas de cultivo a través de la GUI en la Raspberry Pi, generando una solución de bajo costo para el acceso de pequeños y medianos productores en zonas rurales de Colombia., Objective: This paper presents a detailed selection of IoT technologies for the design of an ambient humidity monitoring infrastructure to control crop irrigation. Methodology: The prototype design was supported by previous studies carried out by the authors and tests in 2019 at Finca el Porvenir in Cundinamarca, Colombia, in order to determine the best option to implement in the field through the use of Python software, and elements such as Raspberry Pi and the TTGO development board. Results: To determine the functionality of the software and hardware of the system, different static tests of the prototype were carried out by monitoring humidity and ambient temperature by means of the DHT22 sensor, whose information is sent from a transmitter node to a receiver node in a concentrator to later save the information in a database and, finally, to obtain graphs of the behavior of the relative humidity, temperature, and the actuation of the control element (solenoid valve). Conclusions: With this information it is possible to demonstrate the effectiveness of the prototype, the possibility of remote connection with the LoRa protocol and the handling and management of information from different crop areas through the GUI on the Raspberry Pi, generating a low-cost solution for access by small and medium producers in rural areas in Colombia.

Published
2022

7. ESICM LIVES 2016: part one: Milan, Italy. 1-5 October 2016

Author

Bos, L., Schouten, L., van Vught, L., Wiewel, M., Ong, D., Cremer, O., Artigas, A., Martin-Loeches, I., Hoogendijk, A., van der Poll, T., Horn, J., Juffermans, N., Schultz, M., de Prost, N., Pham, T., Carteaux, G., Dessap, A. Mekontso, Brun-Buisson, C., Fan, E., Bellani, G., Laffey, J., Mercat, A., Brochard, L., Maitre, B., Howells, P. A., Thickett, D. R., Knox, C., Park, D. P., Gao, F., Tucker, O., Whitehouse, T., McAuley, D. F., Perkins, G. D., Pham, T., Laffey, J., Bellani, G., Fan, E., Pisani, L., Roozeman, J. P., Simonis, F. D., Giangregorio, A., Schouten, L. R., Van der Hoeven, S. M., Horn, J., Neto, A. Serpa, Festic, E., Dondorp, A. M., Grasso, S., Bos, L. D., Schultz, M. J., Koster-Brouwer, M., Verboom, D., Scicluna, B., van de Groep, K., Frencken, J., Schultz, M., van der Poll, T., Bonten, M., Cremer, O., Ko, J. I., Kim, K. S., Suh, G. J., Kwon, W. Y., Kim, K., Shin, J. H., Ranzani, O. T., Prina, E., Menendez, R., Ceccato, A., Mendez, R., Cilloniz, C., Gabarrus, A., Ferrer, M., Torres, A., Urbano, A., Zhang, L. A., Swigon, D., Pike, F., Parker, R. S., Clermont, G., Scheer, C., Kuhn, S. O., Modler, A., Vollmer, M., Fuchs, C., Hahnenkamp, K., Rehberg, S., Gründling, M., Taggu, A., Darang, N., Öveges, N., László, I., Tánczos, K., Németh, M., Lebák, G., Tudor, B., Érces, D., Kaszaki, J., Huber, W., Trásy, D., Molnár, Z., Ferrara, G., Edul, V. S. Kanoore, Canales, H. S., Martins, E., Canullán, C., Murias, G., Pozo, M. O., Eguillor, J. F. Caminos, Buscetti, M. G., Ince, C., Dubin, A., Aya, H. D., Rhodes, A., Fletcher, N., Grounds, R. M., Cecconi, M., Jacquet-Lagrèze, M., Riche, M., Schweizer, R., Portran, P., Fornier, W., Lilot, M., Neidecker, J., Fellahi, J. L., Escoresca-Ortega, A., Gutiérrez-Pizarraya, A., Charris-Castro, L., Corcia-Palomo, Y., Fernandez-Delgado, E., Garnacho-Montero, J., Roger, C., Muller, L., Elotmani, L., Lipman, J., Lefrant, J. Y., Roberts, J. A., Muñoz-Bermúdez, R., Samper, M., Climent, C., Vasco, F., Sara, V., Luque, S., Campillo, N., Cerrato, S. Grau, Masclans, J. R., Alvarez-Lerma, F., Brugger, S. Carvalho, Jimenez, G. Jimenez, Torner, M. Miralbés, Cabello, J. Trujillano, Garrido, B. Balsera, Casals, X. Nuvials, Gaite, F. Barcenilla, Vidal, M. Vallverdú, Martínez, M. Palomar, Gusarov, V., Shilkin, D., Dementienko, M., Nesterova, E., Lashenkova, N., Kuzovlev, A., Zamyatin, M., Demoule, A., Carreira, S., Lavault, S., Palancca, O., Morawiec, E., Mayaux, J., Arnulf, I., Similowski, T., Rasmussen, B. S., Maltesen, R. G., Hanifa, M., Pedersen, S., Kristensen, S. R., Wimmer, R., Panigada, M., Bassi, G. Li, Ranzani, O. T., Kolobow, T., Zanella, A., Cressoni, M., Berra, L., Parrini, V., Kandil, H., Salati, G., Livigni, S., Amatu, A., Andreotti, A., Tagliaferri, F., Moise, G., Mercurio, G., Costa, A., Vezzani, A., Lindau, S., Babel, J., Cavana, M., Consonni, D., Pesenti, A., Gattinoni, L., Torres, A., Mansouri, P., Zand, F., Zahed, L., Dehghanrad, F., Bahrani, M., Ghorbani, M., Cambiaghi, B., Moerer, O., Mauri, T., Kunze-Szikszay, N., Ritter, C., Pesenti, A., Quintel, M., Vilander, L. M., Kaunisto, M. A., Vaara, S. T., Pettilä, V., Mulier, J. L. G. Haitsma, Rozemeijer, S., Spoelstra-de Man, A. M. E., Elbers, P. E., Tuinman, P. R., de Waard, M. C., Oudemans-van Straaten, H. M., Liberatore, A. M. A., Souza, R. B., Martins, A. M. C. R. P. F., Vieira, J. C. F., Koh, I. H. J., Martínez, M. Galindo, Sánchez, R. Jiménez, Gascón, L. Martínez, Mulero, M. D. Rodríguez, Freire, A. Ortín, Muñoz, A. Ojados, Acebes, S. Rebollo, Martínez, Á. Fernández, Aliaga, S. Moreno, Para, L. Herrera, Payá, J. Murcia, Mulero, F. Rodríguez, Guerci, P., Ince, Y., Heeman, P., Ergin, B., Ince, C., Uz, Z., Massey, M., Ince, Y., Papatella, R., Bulent, E., Guerci, P., Toraman, F., Ince, C., Longbottom, E. R., Torrance, H. D., Owen, H. C., Hinds, C. J., Pearse, R. M., O’Dywer, M. J., Trogrlic, Z., van der Jagt, M., Lingsma, H., Ponssen, H. H., Schoonderbeek, J. F., Schreiner, F., Verbrugge, S. J., Duran, S., van Achterberg, T., Bakker, J., Gommers, D. A. M. P. J., Ista, E., Krajčová, A., Waldauf, P., Duška, F., Shah, A., Roy, N., McKechnie, S., Doree, C., Fisher, S., Stanworth, S. J., Jensen, J. F., Overgaard, D., Bestle, M. H., Christensen, D. F., Egerod, I., Pivkina, A., Gusarov, V., Zhivotneva, I., Pasko, N., Zamyatin, M., Jensen, J. F., Egerod, I., Bestle, M. H., Christensen, D. F., Alklit, A., Hansen, R. L., Knudsen, H., Grode, L. B., Overgaard, D., Hravnak, M., Chen, L., Dubrawski, A., Clermont, G., Pinsky, M. R., Parry, S. M., Knight, L. D., Connolly, B. C., Baldwin, C. E., Puthucheary, Z. A., Denehy, L., Hart, N., Morris, P. E., Mortimore, J., Granger, C. L., Jensen, H. I., Piers, R., Van den Bulcke, B., Malmgren, J., Metaxa, V., Reyners, A. K., Darmon, M., Rusinova, K., Talmor, D., Meert, A. P., Cancelliere, L., Zubek, L., Maia, P., Michalsen, A., Decruyenaere, J., Kompanje, E., Vanheule, S., Azoulay, E., Vansteelandt, S., Benoit, D., Van den Bulcke, B., Piers, R., Jensen, H. I., Malmgren, J., Metaxa, V., Reyners, A. K., Darmon, M., Rusinova, K., Talmor, D., Meert, A. P., Cancelliere, L., Zubek, L., Maia, P., Michalsen, A., Decruyenaere, J., Kompanje, E., Vanheule, S., Azoulay, E., Vansteelandt, S., Benoit, D., Ryan, C., Dawson, D., Ball, J., Noone, K., Aisling, B., Prudden, S., Ntantana, A., Matamis, D., Savvidou, S., Giannakou, M., Gouva, M., Nakos, G., Koulouras, V., Aron, J., Lumley, G., Milliken, D., Dhadwal, K., McGrath, B. A., Lynch, S. J., Bovento, B., Sharpe, G., Grainger, E., Pieri-Davies, S., Wallace, S., McGrath, B., Lynch, S. J., Bovento, B., Grainger, E., Pieri-Davies, S., Sharpe, G., Wallace, S., Jung, M., Cho, J., Park, H., Suh, G., Kousha, O., Paddle, J., Gripenberg, L. Gamrin, Rehal, M. Sundström, Wernerman, J., Rooyackers, O., de Grooth, H. J., Choo, W. P., Spoelstra-de Man, A. M., Swart, E. L., Oudemans-van Straaten, H. M., Talan, L., Güven, G., Altıntas, N. D., Padar, M., Uusvel, G., Starkopf, L., Starkopf, J., Blaser, A. Reintam, Kalaiselvan, M. S., Arunkumar, A. S., Renuka, M. K., Shivkumar, R. L., Volbeda, M., ten Kate, D., Hoekstra, M., van der Maaten, J. M., Nijsten, M. W., Komaromi, A., Rooyackers, O., Wernerman, J., Norberg, Å., Smedberg, M., Mori, M., Pettersson, L., Norberg, Å., Rooyackers, O., Wernerman, J., Theodorakopoulou, M., Christodoulopoulou, T., Diamantakis, A., Frantzeskaki, F., Kontogiorgi, M., Chrysanthopoulou, E., Lygnos, M., Diakaki, C., Armaganidis, A., Gundogan, K., Dogan, E., Coskun, R., Muhtaroglu, S., Sungur, M., Ziegler, T., Guven, M., Kleyman, A., Khaliq, W., Andreas, D., Singer, M., Meierhans, R., Schuepbach, R., De Brito-Ashurst, I., Zand, F., Sabetian, G., Nikandish, R., Hagar, F., Masjedi, M., Maghsudi, B., Vazin, A., Ghorbani, M., Asadpour, E., Kao, K. C., Chiu, L. C., Hung, C. Y., Chang, C. H., Li, S. H., Hu, H. C., El Maraghi, S., Ali, M., Rageb, D., Helmy, M., Marin-Corral, J., Vilà, C., Masclans, J. R., Vàzquez, A., Martín-Loeches, I., Díaz, E., Yébenes, J. C., Rodriguez, A., Álvarez-Lerma, F., Varga, N., Cortina-Gutiérrez, A., Dono, L., Martínez-Martínez, M., Maldonado, C., Papiol, E., Pérez-Carrasco, M., Ferrer, R., Nweze, K., Morton, B., Welters, I., Houard, M., Voisin, B., Ledoux, G., Six, S., Jaillette, E., Nseir, S., Romdhani, S., Bouneb, R., Loghmari, D., Aicha, N. Ben, Ayachi, J., Meddeb, K., Chouchène, I., Khedher, A., Boussarsar, M., Chan, K. S., Yu, W. L., Marin-Corral, J., Vilà, C., Masclans, J. R., Nolla, J., Vidaur, L., Bonastre, J., Suberbiola, B., Guerrero, J. E., Rodriguez, A., Coll, N. Ramon, Jiménez, G. Jiménez, Brugger, S. Carvalho, Calero, J. Codina, Garrido, B. Balsera, García, M., Martínez, M. Palomar, Vidal, M. Vallverdú, de la Torre, M. C., Vendrell, E., Palomera, E., Güell, E., Yébenes, J. C., Serra-Prat, M., Bermejo-Martín, J. F., Almirall, J., Tomas, E., Escoval, A., Froe, F., Pereira, M. H. Vitoria, Velez, N., Viegas, E., Filipe, E., Groves, C., Reay, M., Chiu, L. C., Hu, H. C., Hung, C. Y., Chang, C. H., Li, S. H., Kao, K. C., Ballin, A., Facchin, F., Sartori, G., Zarantonello, F., Campello, E., Radu, C. M., Rossi, S., Ori, C., Simioni, P., Umei, N., Shingo, I., Santos, A. C., Candeias, C., Moniz, I., Marçal, R., e Silva, Z. Costa, Ribeiro, J. M., Georger, J. F., Ponthus, J. P., Tchir, M., Amilien, V., Ayoub, M., Barsam, E., Martucci, G., Panarello, G., Tuzzolino, F., Capitanio, G., Ferrazza, V., Carollo, T., Giovanni, L., Arcadipane, A., Sánchez, M. López, González-Gay, M. A., Díaz, F. J. Llorca, López, M. I. Rubio, Zogheib, E., Villeret, L., Nader, J., Bernasinski, M., Besserve, P., Caus, T., Dupont, H., Morimont, P., Habran, S., Hubert, R., Desaive, T., Blaffart, F., Janssen, N., Guiot, J., Pironet, A., Dauby, P., Lambermont, B., Zarantonello, F., Ballin, A., Facchin, F., Sartori, G., Campello, E., Pettenuzzo, T., Citton, G., Rossi, S., Simioni, P., Ori, C., Kirakli, C., Ediboglu, O., Ataman, S., Yarici, M., Tuksavul, F., Keating, S., Gibson, A., Gilles, M., Dunn, M., Price, G., Young, N., Remeta, P., Bishop, P., Zamora, M. D. Fernández, Muñoz-Bono, J., Curiel-Balsera, E., Aguilar-Alonso, E., Hinojosa, R., Gordillo-Brenes, A., Arboleda-Sánchez, J. A., Skorniakov, I., Vikulova, D., Whiteley, C., Shaikh, O., Jones, A., Ostermann, M., Forni, L., Scott, M., Sahatjian, J., Linde-Zwirble, W., Hansell, D., Laoveeravat, P., Srisawat, N., Kongwibulwut, M., Peerapornrattana, S., Suwachittanont, N., Wirotwan, T. O., Chatkaew, P., Saeyub, P., Latthaprecha, K., Tiranathanagul, K., Eiam-ong, S., Kellum, J. A., Berthelsen, R. E., Perner, A., Jensen, A. E. K., Jensen, J. U., Bestle, M. H., Gebhard, D. J., Price, J., Kennedy, C. E., Akcan-Arikan, A., Liberatore, A. M. A., Souza, R. B., Martins, A. M. C. R. P. F., Vieira, J. C. F., Kang, Y. R., Nakamae, M. N., Koh, I. H. J., Hamed, K., Khaled, M. M., Soliman, R. Aly, Mokhtar, M. Sherif, Seller-Pérez, G., Arias-Verdú, D., Llopar-Valdor, E., De-Diós-Chacón, I., Quesada-García, G., Herrera-Gutierrez, M. E., Hafes, R., Carroll, G., Doherty, P., Wright, C., Vera, I. G. Guerra, Ralston, M., Gemmell, M. L., MacKay, A., Black, E., Wright, C., Docking, R. I., Appleton, R., Ralston, M. R., Gemmell, L., Appleton, R., Wright, C., Docking, R. I., Black, E., Mackay, A., Rozemeijer, S., Mulier, J. L. G. Haitsma, Röttgering, J. G., Elbers, P. W. G., Spoelstra-de Man, A. M. E., Tuinman, P. R., de Waard, M. C., Oudemans-van Straaten, H. M., Mejeni, N., Nsiala, J., Kilembe, A., Akilimali, P., Thomas, G., Egerod, I., Andersson, A. E., Fagerdahl, A. M., Knudsen, V., Meddeb, K., Cheikh, A. Ben, Hamdaoui, Y., Ayachi, J., Guiga, A., Fraj, N., Romdhani, S., Sma, N., Bouneb, R., Chouchene, I., Khedher, A., Bouafia, N., Boussarsar, M., Amirian, A., Ziaian, B., Masjedi, M., Fleischmann, C., Thomas-Rueddel, D. O., Schettler, A., Schwarzkopf, D., Stacke, A., Reinhart, K., Filipe, E., Escoval, A., Martins, A., Sousa, P., Velez, N., Viegas, E., Tomas, E., Snell, G., Matsa, R., Paary, T. T. S., Kalaiselvan, M. S., Cavalheiro, A. M., Rocha, L. L., Vallone, C. S., Tonilo, A., Lobato, M. D. S., Malheiro, D. T., Sussumo, G., Lucino, N. M., Zand, F., Rosenthal, V. D., Masjedi, M., Sabetian, G., Maghsudi, B., Ghorbani, M., Dashti, A. Sanaei, Yousefipour, A., Goodall, J. R., Williamson, M., Tant, E., Thomas, N., Balci, C., Gonen, C., Haftacı, E., Gurarda, H., Karaca, E., Paldusová, B., Zýková, I., Šímová, D., Houston, S., D’Antona, L., Lloyd, J., Garnelo-Rey, V., Sosic, M., Sotosek-Tokmazic, V., Kuharic, J., Antoncic, I., Dunatov, S., Sustic, A., Chong, C. T., Sim, M., Lyovarin, T., Díaz, F. M. Acosta, Galdó, S. Narbona, Garach, M. Muñoz, Romero, O. Moreno, Bailón, A. M. Pérez, Pinel, A. Carranza, Colmenero, M., Gritsan, A., Gazenkampf, A., Korchagin, E., Dovbish, N., Lee, R. M., Lim, M. P. P., Chong, C. T., Lim, B. C. L., See, J. J., Assis, R., Filipe, F., Lopes, N., Pessoa, L., Pereira, T., Catorze, N., Aydogan, M. S., Aldasoro, C., Marchio, P., Jorda, A., Mauricio, M. D., Guerra-Ojeda, S., Gimeno-Raga, M., Colque-Cano, M., Bertomeu-Artecero, A., Aldasoro, M., Valles, S. L., Tonon, D., Triglia, T., Martin, J. C., Alessi, M. C., Bruder, N., Garrigue, P., Velly, L., Spina, S., Scaravilli, V., Marzorati, C., Colombo, E., Savo, D., Vargiolu, A., Cavenaghi, G., Citerio, G., Andrade, A. H. V., Bulgarelli, P., Araujo, J. A. P., Gonzalez, V., Souza, V. A., Costa, A., Massant, C., Filho, C. A. C. Abreu, Morbeck, R. A., Burgo, L. E., van Groenendael, R., van Eijk, L. T., Leijte, G. P., Koeneman, B., Kox, M., Pickkers, P., García-de la Torre, A., de la Torre-Prados, M., Fernández-Porcel, A., Rueda-Molina, C., Nuevo-Ortega, P., Tsvetanova-Spasova, T., Cámara-Sola, E., García-Alcántara, A., Salido-Díaz, L., Liao, X., Feng, T., Zhang, J., Cao, X., Wu, Q., Xie, Z., Li, H., Kang, Y., Winkler, M. S., Nierhaus, A., Mudersbach, E., Bauer, A., Robbe, L., Zahrte, C., Schwedhelm, E., Kluge, S., Zöllner, C., Morton, B., Mitsi, E., Pennington, S. H., Reine, J., Wright, A. D., Parker, R., Welters, I. D., Blakey, J. D., Rajam, G., Ades, E. W., Ferreira, D. M., Wang, D., Kadioglu, A., Gordon, S. B., Koch, R., Kox, M., Rahamat-Langedoen, J., Schloesser, J., de Jonge, M., Pickkers, P., Bringue, J., Guillamat-Prats, R., Torrents, E., Martinez, M. L., Camprubí-Rimblas, M., Artigas, A., Blanch, L., Park, S. Y., Park, Y. B., Song, D. K., Shrestha, S., Park, S. H., Koh, Y., Park, M. J., Hong, C. W., Lesur, O., Coquerel, D., Sainsily, X., Cote, J., Söllradl, T., Murza, A., Dumont, L., Dumaine, R., Grandbois, M., Sarret, P., Marsault, E., Salvail, D., Auger-Messier, M., Chagnon, F., Lauretta, M. P., Greco, E., Dyson, A., Singer, M., Preau, S., Ambler, M., Sigurta, A., Saeed, S., Singer, M., Sarıca, L. Topcu, Zibandeh, N., Genc, D., Gul, F., Akkoc, T., Kombak, E., Cinel, L., Akkoc, T., Cinel, I., Pollen, S. J., Arulkumaran, N., Singer, M., Torrance, H. D., Longbottom, E. R., Warnes, G., Hinds, C. J., Pennington, D. J., Brohi, K., O’Dwyer, M. J., Kim, H. Y., Na, S., Kim, J., Chang, Y. F., Chao, A., Shih, P. Y., Lee, C. T., Yeh, Y. C., Chen, L. 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R., Parenti, N., Agrusta, F., Palazzi, C., Pifferi, B., Sganzerla, R., Tagliazucchi, F., Luciani, A., Möller, M., Müller-Engelmann, J., Montag, G., Adams, P., Lange, C., Neuzner, J., Gradaus, R., Wodack, K. H., Thürk, F., Waldmann, A. D., Grässler, M. F., Nishimoto, S., Böhm, S. H., Kaniusas, E., Reuter, D. A., Trepte, C. J., Sigmundsson, T., Öhman, T., Redondo, E., Hallbäck, M., Wallin, M., Sipman, F. Suarez, Oldner, A., Sander, C. Hällsjö, Björne, H., Colinas, L., Hernandez, G., Vicho, R., Serna, M., Cuena, R., Canabal, A., Chaari, A., Hakim, K. Abdel, Etman, M., El Bahr, M., El Sakka, A., Bousselmi, K., Arali, A., Kauts, V., Casey, W. F., Bond, O., De Santis, P., Iesu, E., Franchi, F., Vincent, J. L., Creteur, J., Scolletta, S., Taccone, F. S., Marutyan, Z., Hamidova, L., Shakotko, A., Movsisyan, V., Uysupova, I., Evdokimov, A., Petrikov, S., Gonen, C., Haftacı, E., Balci, C., Calvo, F. J. Redondo, Bejarano, N., Baladron, V., Villazala, R., Redondo, J., Padilla, D., Villarejo, P., Akcan-Arikan, A., Kennedy, C. E., Arzapalo, M. F. Aguilar, Gomez-Gonzalez, C., Mas-Font, S., Puppo-Moreno, A., Herrera-Gutierrez, M., Garcia-Garcia, M., Aldunate-Calvo, S., Plata-Menchaca, E. P., Pérez-Fernández, X. L., Estruch, M., Betbese-Roig, A., Campos, P. Cárdenas, Lora, M. Rojas, Gaibor, N. D. Toapanta, Medina, R. S. Contreras, Sanguino, V. D. Gumucio, Casanova, E. J., Riera, J. Sabater, Kritmetapak, K., Peerapornratana, S., Kittiskulnam, P., Dissayabutra, T., Tiranathanagul, K., Susantithapong, P., Praditpornsilpa, K., Tungsanga, K., Eiam-Ong, S., Srisawat, N., Winkelmann, T., Busch, T., Meixensberger, J., Bercker, S., Cabeza, E. M. Flores, Sánchez, M. Sánchez, Giménez, N. Cáceres, Melón, C. Gutierrez, de Lucas, E. Herrero, Estañ, P. Millán, Bernal, M. Hernández, de Lorenzo y Mateos, A. Garcia, Ergin, B., Guerci, P., Specht, P. A. C., Ince, Y., Ince, C., Balik, M., Zakharchenko, M., Los, F., Brodska, H., de Tymowski, C., Augustin, P., Desmard, M., Montravers, P., Stapel, S. N., de Boer, R., Oudemans, H. M., Hollinger, A., Schweingruber, T., Jockers, F., Dickenmann, M., Siegemund, M., Runciman, N., Ralston, M., Appleton, R., Mauri, T., Alban, L., Turrini, C., Sasso, T., Langer, T., Panigada, M., Taccone, P., Carlesso, E., Marenghi, C., Grasselli, G., Pesenti, A., Wibart, P., Reginault, T., Garcia, M., Barbrel, B., Benard, A., Bader, C., Vargas, F., Bui, H. N., Hilbert, G., Simón, J. M. Serrano, Sánchez, P. Carmona, Ferrón, F. Ruiz, de Acilu, M. García, Marin, J., Antonia, V., Ruano, L., Monica, M., Ferrer, R., Masclans, J. R., Roca, O., Hong, G., Kim, D. H., Kim, Y. S., Park, J. S., Jee, Y. K., xiang, Z. Yu, Jia-xing, W., dan, W. Xiao, long, N. Wen, Yu, W., Yan, Z., Cheng, X., Kobayashi, T., Onodera, Y., Akimoto, R., Sugiura, A., Suzuki, H., Iwabuchi, M., Nakane, M., Kawamae, K., Sanchez, P. Carmona, Rodriguez, M. D. Bautista, Delgado, M. Rodriguez, Sánchez, V. Martínez de Pinillos, Gómez, A. Mula, Simón, J. M. Serrano, Beuret, P., Fortes, C., Lauer, M., Reboul, M., Chakarian, J. C., Fabre, X., Philippon-Jouve, B., Devillez, S., Clerc, M., Rittayamai, N., Sklar, M., Dres, M., Rauseo, M., Campbell, C., West, B., Tullis, D. E., Brochard, L., Onodera, Y., Akimoto, R., Suzuki, H., Okada, M., Nakane, M., Kawamae, K., Ahmad, N., Wood, M., Glossop, A., Lucas, J. Higuera, Ortiz, A. Blandino, Alonso, D. Cabestrero, De Pablo Sánchez, R., González, L. Rey, Costa, R., Spinazzola, G., Pizza, A., Ferrone, G., Rossi, M., Antonelli, M., Conti, G., Ribeiro, H., Alves, J., Sousa, M., Reis, P., Socolovsky, C. S., Cauley, R. P., Frankel, J. E., Beam, A. L., Olaniran, K. O., Gibbons, F. K., Christopher, K. B., Pennington, J., Zolfaghari, P., King, H. S., Kong, H. H. Y., Shum, H. P., Yan, W. W., Kaymak, C., Okumus, N., Sari, A., Erdogdu, B., Aksun, S., Basar, H., Ozcan, A., Ozcan, N., Oztuna, D., Malmgren, J. A., Lundin, S., Torén, K., Eckerström, M., Wallin, A., Waldenström, A. C., Riccio, F. C., Pogson, D., Antonio, A. C. P., Leivas, A. F., Kenji, F., James, E., Morgan, P., Carroll, G., Gemmell, L., MacKay, A., Wright, C., Ballantyne, J., Jonnada, S., Gerrard, C. S., Jones, N., Salciccioli, J. D., Marshall, D. C., Komorowski, M., Hartley, A., Sykes, M. C., Goodson, R., Shalhoub, J., Villanueva, J. R. Fernández, Garda, R. Fernández, Lago, A. M. López, Ruiz, E. Rodríguez, Vaquero, R. Hernández, Rodríguez, C. Galbán, Pérez, E. Varo, Hilasque, C., Oliva, I., Sirgo, G., Martin, M. C., Olona, M., Gilavert, M. C., Bodí, M., Ebm, C., Aggarwal, G., Huddart, S., Quiney, N., Cecconi, M., Fernandes, S. M., Silva, J. Santos, Gouveia, J., Silva, D., Marques, R., Bento, H., Alvarez, A., Silva, Z. Costa, Diaz, D. Díaz, Martínez, M. Villanova, Herrejon, E. Palencia, de la Gandara, A. Martinez, Gonzalo, G., Lopez, M. A., de Gopegui Miguelena, P. Ruíz, Matilla, C. I. Bernal, Chueca, P. Sánchez, Longares, M. D. C. Rodríguez, Abril, R. Ramos, Aguilar, A. L. Ruíz, de Murillas, R. Garrido López, Fernández, R. Fernández, Laborías, P. Morales, Castellanos, M. A. Díaz, Laborías, M. E. Morales, Cho, J., Kim, J., Park, J., Woo, S., West, T., Powell, E., Rimmer, A., Orford, C., Jones, N., Williams, J., Matilla, C. I. Bernal, de Gopegui Miguelena, P. Ruiz, Chueca, P. Sánchez, Abril, R. Ramos, Longares, M. D. C. Rodríguez, Aguilar, A. L. Ruíz, de Murillas, R. Garrido López, Bourne, R. S., Shulman, R., Tomlin, M., Mills, G. H., Borthwick, M., Berry, W., Huertas, D. García, Manzano, F., Villagrán-Ramírez, F., Ruiz-Perea, A., Rodríguez-Mejías, C., Santiago-Ruiz, F., Colmenero-Ruiz, M., König, C., Matt, B., Kortgen, A., Hartog, C. S., Wong, A., Balan, C., Barker, G., Srisawat, N., Peerapornratana, S., Laoveeravat, P., Tachaboon, S., Eiam-ong, S., Paratz, J., Kayambu, G., Boots, R., Arzapalo, M. F. Aguilar, Vlasenko, R., Gromova, E., Loginov, S., Kiselevskiy, M., Dolgikova, Y., Tang, K. B., Chau, C. M., Lam, K. N., Gil, E., Suh, G. Y., Park, C. M., Park, J., Chung, C. R., Lee, C. T., Chao, A., Shih, P. Y., Chang, Y. F., Lai, C. H., Hsu, Y. C., Yeh, Y. C., Cheng, Y. J., Colella, V., Zarrillo, N., D’Amico, M., Forfori, F., Pezza, B., Laddomada, T., Beltramelli, V., Pizzaballa, M. L., Doronzio, A., Balicco, B., Kiers, D., van der Heijden, W., Gerretsen, J., de Mast, Q., el Messaoudi, S., Rongen, G., Gomes, M., Kox, M., Pickkers, P., Riksen, N. P., Kashiwagi, Y., Okada, M., Hayashi, K., Inagaki, Y., Fujita, S., Nakamae, M. N., Kang, Y. R., Souza, R. B., Liberatore, A. M. A., Koh, I. H. J., Blet, A., Sadoune, M., Lemarié, J., Bihry, N., Bern, R., Polidano, E., Merval, R., Launay, J. M., Lévy, B., Samuel, J. L., Mebazaa, A., Hartmann, J., Harm, S., and Weber, V.

Published
2016
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8. Diagnóstico molecular de SARS-CoV-2

Author

Pinilla B, Gladys, Cruz B, Claudia Andrea, and Navarrete O, Jeannette

Subjects
diagnosis, virus SARS, coronavirus, SARS virus, RT qPCR, diagnóstico
Abstract

Resumen El diagnóstico de COVID-19 se basa tanto en aspectos clínicos como en pruebas de detección, pero los síntomas y signos clínicos de los pacientes infectados son altamente atípicos y, por lo tanto, las pruebas moleculares son indispensables para su diagnóstico. La reacción en cadena de la polimerasa con transcriptasa inversa (RT-qPCR) se lleva a cabo en laboratorios nivel BSL II; asimismo, los principales blancos moleculares para la detección viral son el gen E (envoltura), y el gen RdRP (ARN polimerasa dependiente de ARN). Los falsos negativos en este diagnóstico se deben a la calidad y cantidad de la muestra, condiciones de transporte, almacenamiento, y manejo de estas antes y después de la extracción (el ARN es termolábil y abundantes las RNasas), fase de la infección, mutaciones del virus y presencia de inhibidores de la PCR. En estos casos, se recomienda una nueva toma de muestra, especialmente de vías respiratorias bajas, para aumentar la carga viral. Se debe tener en cuenta la sensibilidad analítica de la RT-qPCR (5,2 copias de ARN/reacción) y que, una vez el RNA se extrae, se va degradando progresivamente afectando la sensibilidad diagnóstica de la prueba. Un diagnóstico oportuno permite optimizar el manejo (aislamiento y tratamiento) y monitorización de los pacientes, así como la aplicación de medidas de prevención y control de la expansión, y la vigilancia epidemiológica de la enfermedad. ABSTRACT COVID-19 diagnosis is based on both clinical aspects and screening tests. However, clinical symptoms and signs in infected patients are highly atypical; hence, molecular tests are essential for diagnosis. RT-qPCR is carried out at BSL II level laboratories; the main molecular targets for viral detection are E gene (envelope), and RdRP gene (RNA-dependent RNA polymerase). False negatives in this diagnosis are due to sample quality and quantity, transport conditions, storage and handling before and after extraction (RNA is heat-labile and RNases are abundant); infection phase; virus mutations and presence of CRP inhibitors. Taking into account analytical sensitivity of RT-qPCR (5.2 copies of RNA / reaction) and the fact that once RNA it is extracted, it progressively degrades and affects test diagnostic sensitivity, a new sample -specifically taken from the lower respiratory tract in order to increase viral load- is recommended in the abovementioned cases. Timely diagnosis allows optimizing management (isolation and treatment), patient monitoring, implementing prevention and control measures as well as epidemiological surveillance of the disease.

Published
2021

10. Design proposal of a wireless sensor and actuator network for irrigation, based on Internet of Things technology.

Author

Tovar Soto, J. P., García Navarrete, O. L., and Gutiérrez-Martínez, L. C.

Subjects
DATABASES, WIRELESS sensor networks, INTERNET of things, WIRELESS communications, PYTHON programming language, IRRIGATION, RASPBERRY Pi, HUMIDITY
Abstract

Copyright of Investigación e Innovación en Ingenierías is the property of Universidad Simon Bolivar and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Published
2022
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11. Determinación de los genes, 16S ADNr, polA, y TpN47, en la detección de Treponema pallidum subsp. pallidum para el diagnóstico de sífilis congénita

Author

Pinilla B, Gladys, Chavarro P, Bibiana, Moreno A, Natalia, Navarrete O, Jeannette, and Muñoz M, Liliana

Subjects
nested PCR, Sífilis congénita, PCR en Tiempo Real, Treponema pallidum subsp. pallidum, Congenital syphilis, PCR anidada, real-time PCR
Abstract

Objetivo. Comparar el comportamiento de tres genes diana 16S ADNr, polA, y TpN47, para la detección de T. pallidum subsp. pallidum. Métodos. Se usaron técnicas moleculares como la reacción en cadena de la polimerasa en muestras de cordón umbilical. Mediante PCR convencional, PCR anidada y PCR en tiempo real se amplificaron blancos moleculares del microorganismo. Resultados. Se evidenció que con los tres genes por PCR convencional se obtienen similares resultados, pero por con PCR anidada y PCR en Tiempo Real, el gen TpN47 tiene mayor sensibilidad en comparación con los genes polA y 16S ADNr. Se concluye que el gen TpN47 se puede usar como blanco molecular para el diagnóstico oportuno de sífilis congénita por medio de PCR anidada y en tiempo real, ya que alcanzó la máxima sensibilidad y especificidad en este estudio. Objective. To compare the behavior of the three target genes (16S rDNA, polA, and TpN47) for the detection of T. pallidum subsp. Pallidum. Methods. Molecular techniques such as polymerase chain reaction were used on samples of an umbilical cord. Molecular targets of the microorganism were amplified by means of conventional PCR, nested PCR and real-time PCR. Results. Similar results for the three genes were obtained by conventional PCR; but in the case of nested PCR and real-time PCR, the gen TpN47 exhibited higher sensitivity in comparison to the genes polA and 16S rDNA. In conclusion, the gen TpN47 can be used as a molecular target for the prompt diagnosis of congenital syphilis through nested PCR and real-time PCR due to its high sensitivity and specificity shown in this study.

Published
2015

14. Macrophages allocate before apoptosis initiation and produce reactive oxygen species during interdigital phagocytosis.

Author

Hernández-García D, García-Meléndrez C, Hernández-Martínez R, Collazo-Navarrete O, and Covarrubias L

Subjects
Animals, Mice, NADPH Oxidases metabolism, Reactive Oxygen Species metabolism, Phagocytosis, Apoptosis, Macrophages metabolism, Lysosomes metabolism
Abstract

During programmed cell death (PCD), it is commonly accepted that macrophages are recruited by apoptotic cells to complete cell degradation. Interdigital cell death, a classical model of PCD, contributes to digit individualization in limbs of mammals and other vertebrates. Here, we show that macrophages are present in interdigits before significant cell death occurs and remain after apoptosis inhibition. The typical interdigital phagocytic activity was not observed after a partial depletion of macrophages and was markedly reduced by engulfment/phagosome maturation inhibition, as detected by its association with high lysosomal activity. β-galactosidase activity in this region was also coupled with phagocytosis, against its relationship with cellular senescence. Interdigital phagocytosis correlated with high levels of reactive oxygen species (ROS), common in embryo regions carrying abundant cell death, suggesting that macrophages are the major source of ROS. ROS generation was dependent on NADPH oxidases and blood vessel integrity, but not directly associated with lysosomal activity. Therefore, macrophages prepattern regions where abundant cell death is going to occur, and high lysosomal activity and the generation of ROS by an oxidative burst-like phenomenon are activities of phagocytosis., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2024. Published by The Company of Biologists Ltd.)

Published
2024
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15. Alpha-Synuclein Gene Alterations Modulate Tyrosine Hydroxylase in Human iPSC-Derived Neurons in a Parkinson's Disease Animal Model.

Author

Bernal-Conde LD, Peña-Martínez V, Morato-Torres CA, Ramos-Acevedo R, Arias-Carrión Ó, Padilla-Godínez FJ, Delgado-González A, Palomero-Rivero M, Collazo-Navarrete O, Soto-Rojas LO, Gómez-Chavarín M, Schüle B, and Guerra-Crespo M

Abstract

Parkinson's disease (PD) caused by SNCA gene triplication (3X SNCA ) leads to early onset, rapid progression, and often dementia. Understanding the impact of 3X SNCA and its absence is crucial. This study investigates the differentiation of human induced pluripotent stem cell (hiPSC)-derived floor-plate progenitors into dopaminergic neurons. Three different genotypes were evaluated in this study: patient-derived hiPSCs with 3X SNCA , a gene-edited isogenic line with a frame-shift mutation on all SNCA alleles ( SNCA 4KO), and a normal wild-type control. Our aim was to assess how the substantia nigra pars compacta (SNpc) microenvironment, damaged by 6-hydroxydopamine (6-OHDA), influences tyrosine hydroxylase-positive (Th+) neuron differentiation in these genetic variations. This study confirms successful in vitro differentiation into neuronal lineage in all cell lines. However, the SNCA 4KO line showed unusual LIM homeobox transcription factor 1 alpha (Lmx1a) extranuclear distribution. Crucially, both 3X SNCA and SNCA 4KO lines had reduced Th+ neuron expression, despite initial successful neuronal differentiation after two months post-transplantation. This indicates that while the SNpc environment supports early neuronal survival, SNCA gene alterations-either amplification or knock-out-negatively impact Th+ dopaminergic neuron maturation. These findings highlight SNCA 's critical role in PD and underscore the value of hiPSC models in studying neurodegenerative diseases.

Published
2024
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16. Congenital diaphragmatic hernia: phosphodiesterase-5 and Arginase inhibitors prevent pulmonary vascular hypoplasia in rat lungs.

Author

Toso A, Aránguiz O, Céspedes C, Navarrete O, Hernández C, Vio CP, Luco M, Casanello P, and Kattan J

Subjects
Rats, Female, Pregnancy, Animals, Cyclic Nucleotide Phosphodiesterases, Type 5 adverse effects, Arginase, Sildenafil Citrate pharmacology, Sildenafil Citrate therapeutic use, Rats, Sprague-Dawley, Lung, Phenyl Ethers pharmacology, Disease Models, Animal, Hernias, Diaphragmatic, Congenital pathology
Abstract

Background: Severe pulmonary hypoplasia related to congenital diaphragmatic hernia (CDH) continues to be a potentially fatal condition despite advanced postnatal management strategies., Objective: To evaluate the effect of the antenatal sildenafil and 2(S)-amino-6-boronohexanoic acid (ABH-Arginase inhibitor) on lung volume, pulmonary vascular development, and nitric oxide (NO) synthesis in a Nitrofen-induced CDH rat model., Methods: Nitrofen-induced CDH rat model was used. Nitrofen was administrated on embryonic day(E) 9,5. At E14, five intervention groups were treated separately: Nitrofen, Nitrofen+Sildenafil, Nitrofen+ABH, Nitrofen+Sildenafil+ABH and Control. At term, offspring's lungs were weighed, some paraffin-embedded for histology, others snap-frozen to analyze eNOS, Arginase I-II expression, and activity., Results: In CDH-bearing offsprings, ABH or Sildenafil+ABH preserved the total lung/body-weight index (p < 0.001), preventing pulmonary vascular smooth muscle cell hyperproliferation and improving lung morphometry. Sildenafil+ABH increased 1.7-fold the lung nitrite levels (p < 0.01) without changes in eNOS expression. Sildenafil and ABH improved the number of pulmonary vessels., Conclusion: These results suggest that in this CDH rat model, the basal activity of Arginase participates in the lung volume and, together with phosphodiesterase-5, regulates NOS activity in the term fetal lung. The combined treatment (Sildenafil+ABH) could revert some of the pulmonary features in CDH by improving the local NO synthesis and preventing smooth muscle cell hyperproliferation., Impact: This study presents Arginase inhibition as a new therapeutic target and the importance of the combined antenatal treatment to improve pulmonary vascular development in a congenital diaphragmatic hernia (CDH) rat model. This study shows that the action of an Arginase inhibitor (ABH) enhances the effects already described for sildenafil in this model. These results reinforce the importance of prenatal treatments' synergy in recovering the hypoplastic lung in the Nitrofen-induced CDH rat model., (© 2022. The Author(s), under exclusive licence to the International Pediatric Research Foundation, Inc.)

Published
2024
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17. Bacterially enhanced plant-growing media for controlled environment agriculture.

Author

Van Gerrewey T, Navarrete O, Vandecruys M, Perneel M, Boon N, and Geelen D

Subjects
RNA, Ribosomal, 16S genetics, Plants genetics, Soil chemistry, Plant Roots microbiology, Soil Microbiology, Agriculture, Bacteria genetics
Abstract

Microbe-plant interactions in the root zone not only shape crop performance in soil but also in hydroponic cultivation systems. The biological and physicochemical properties of the plant-growing medium determine the root-associated microbial community and influence bacterial inoculation effectiveness, which affects plant growth. This study investigated the combined impact of plant-growing media composition and bacterial community inoculation on the root-associated bacterial community of hydroponically grown lettuce (Lactuca sativa L.). Ten plant-growing media were composed of varying raw materials, including black peat, white peat, coir pith, wood fibre, composted bark, green waste compost, perlite and sand. In addition, five different bacterial community inocula (BCI S1-5) were collected from the roots of lettuce obtained at different farms. After inoculation and cultivation inside a vertical farm, lettuce root-associated bacterial community structures, diversity and compositions were determined by evaluating 16S rRNA gene sequences. The study revealed distinct bacterial community structures among experimental replicates, highlighting the influence of raw material variations on root-associated bacterial communities, even at the batch level. However, bacterial community inoculation allowed modulation of the root-associated bacterial communities independently from the plant-growing medium composition. Bacterial diversity was identified as a key determinant of plant growth performance with green waste compost introducing Bacilli and Actinobacteria, and bacterial community inoculum S3 introducing Pseudomonas, which positively correlated with plant growth. These findings challenge the prevailing notion of hydroponic cultivation systems as sterile environments and highlight the significance of proper plant-growing media raw material selection and bacterial community inoculation in shaping root-associated microbiomes that provide stability through microbial diversity. This study supports the concept of creating bacterially enhanced plant-growing media to promote plant growth in controlled environment agriculture., (© 2024 The Authors. Microbial Biotechnology published by Applied Microbiology International and John Wiley & Sons Ltd.)

Published
2024
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18. Embryoid Body Cells from Human Embryonic Stem Cells Overexpressing Dopaminergic Transcription Factors Survive and Initiate Neurogenesis via Neural Rosettes in the Substantia Nigra.

Author

Ramos-Acevedo R, Morato-Torres CA, Padilla-Godínez FJ, Bernal-Conde LD, Palomero-Rivero M, Zafar F, Collazo-Navarrete O, Soto-Rojas LO, Schüle B, and Guerra-Crespo M

Abstract

Transplantation of immature dopaminergic neurons or neural precursors derived from embryonic stem cells (ESCs) into the substantia nigra pars compacta (SNpc) is a potential therapeutic approach for functional restitution of the nigrostriatal pathway in Parkinson's disease (PD). However, further studies are needed to understand the effects of the local microenvironment on the transplanted cells to improve survival and specific differentiation in situ. We have previously reported that the adult SNpc sustains a neurogenic microenvironment. Non-neuralized embryoid body cells (EBCs) from mouse ESCs (mESCs) overexpressing the dopaminergic transcription factor Lmx1a gave rise to many tyrosine hydroxylase (Th + ) cells in the intact and damaged adult SNpc, although only for a short-term period. Here, we extended our study by transplanting EBCs from genetically engineered naive human ESC (hESC), overexpressing the dopaminergic transcription factors LMX1A , FOXA2 , and OTX2 (hESC-LFO), in the SNpc. Unexpectedly, no graft survival was observed in wild-type hESC EBCs transplants, whereas hESC-LFO EBCs showed viability in the SNpc. Interestingly, neural rosettes, a developmental hallmark of neuroepithelial tissue, emerged at 7- and 15-days post-transplantation (dpt) from the hESC-LFO EBCs. Neural rosettes expressed specification dopaminergic markers (Lmx1a, Otx2), which gave rise to several Th + cells at 30 dpt. Our results suggest that the SNpc enables the robust initiation of neural differentiation of transplanted human EBCs prompted to differentiate toward the midbrain dopaminergic phenotype.

Published
2023
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19. Lumbar Facet Effusions and Other Degeneration Parameters and Its Association with Instability.

Author

Bendersky D, Asem M, and Navarrete O

Subjects
Humans, Exudates and Transudates, Retrospective Studies, Spondylolisthesis etiology, Intervertebral Disc Degeneration complications, Intervertebral Disc Degeneration diagnosis, Joint Instability diagnosis, Joint Instability etiology, Lumbar Vertebrae, Zygapophyseal Joint
Abstract

Background: Controversy exists in the literature about whether facet effusions and other degeneration parameters are associated with instability., Objective: To assess the association between facet effusions and other lumbar degeneration parameters and segmental instability., Material and Methods: In this study, 207 L4-L5 and L5-S1 levels in 104 patients were assessed. We divided the spinal levels into two groups: the small facet effusions (SFE) group in whom facet effusions were <1.5 mm or non-effusions were found, and the large facet effusions (LFE) group in whom they were ≥1.5 mm. The association between other degeneration parameters and instability was also assessed, such as disc degeneration, Modic changes (MC), spondylolisthesis, facet orientation and tropism, facet subchondral sclerosis, and facet cartilage degeneration. Furthermore, we subdivided the levels into subgroups to evaluate the association of LFE and instability within each one., Results: The main analysis comparing the presence of instability in SFE and LFE groups showed a non-statistically significant association between LFE and instability. The presence of MC type 1 and the existence of L4-L5 spondylolisthesis had a statistically significant association with instability. In the subset of 43 levels with L4-L5 degenerative spondylolisthesis, the presence of LFE and the existence of MC type 1 reached a significant association with instability., Conclusion: The presence of LFE and/or MC type 1 may act as red flags in patients with L4-L5 degenerative spondylolisthesis to suspect segmental instability., Competing Interests: None

Published
2022
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20. Contrast-enhanced mammography predicts pathological response after neoadjuvant chemotherapy in locally advanced breast cancer.

Author

Canteros D, Walbaum B, Córdova-Delgado M, Torrealba A, Reyes C, Navarro ME, Razmilic D, Camus M, Dominguez F, Navarrete O, Pinto MP, Pizarro G, Acevedo F, and Sánchez C

Abstract

Introduction: Recently, contrast-enhanced mammography (CEM) has emerged as a reliable alternative to breast magnetic resonance imaging (MRI) for the assessment of pathological response in breast cancer patients. Our study sought to determine the diagnostic accuracy of CEM to predict pathological complete response (pCR) in patients who received neoadjuvant chemotherapy (NACT)., Methods: We retrieved the medical records of patients who underwent NACT at our institution. Using post-surgery pCR, morphological evidence and CEM enhancement tumours were classified as follows: 1) radiologic complete response (rCR); 2) functional radiological complete response (frCR); and 3) non-complete response. Initially, we used multivariate analyses adjusted by clinical variables and frCR or rCR to determine which variables affected pathological response. Then, CEM diagnostic accuracy to discriminate pCR was assessed using receiver operating characteristic curves in univariate and multivariate models including either frCR or rCR., Results: A total of 48 patients were included in our study. Most patients (68.7%) were hormone receptor (HR)+ and 41.6% (20) of the patients achieved pCR. Using univariate logistic regression analyses we found that HR status, HER2 status, rCR and frCR had a significant impact on CEM diagnostic accuracy. Exploratory analyses found that CEM sensitivity was higher for HR- tumours. Multivariate logistic regression analyses found 60% sensitivity, 92.9% specificity and 79.2% accuracy in a model that included clinical variables and rCR., Conclusion: CEM is a reliable alternative to high-cost, time-consuming breast MRI that predicts pCR in patients undergoing NACT; CEM diagnostic accuracy was higher among patients who harboured HR- tumours., Competing Interests: None., (© the authors; licensee ecancermedicalscience.)

Published
2022
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21. Embryoid Body Formation from Mouse and Human Pluripotent Stem Cells for Transplantation to Study Brain Microenvironment and Cellular Differentiation.

Author

Guerra-Crespo M, Collazo-Navarrete O, Ramos-Acevedo R, Morato-Torres CA, and Schüle B

Subjects
Animals, Cell Differentiation genetics, Embryoid Bodies, Embryonic Stem Cells, Humans, Mesencephalon, Mice, Induced Pluripotent Stem Cells, Pluripotent Stem Cells
Abstract

Human embryonic stem cell (hESC) and human-induced pluripotent stem cell (hiPSC) technologies have a critical role in regenerative strategies for personalized medicine. Both share the ability to differentiate into almost any cell type of the human body. The study of their properties and clinical applications requires the development of robust and reproducible cell culture paradigms that direct cell differentiation toward a specific phenotype in vitro and in vivo. Our group evaluated the potential of mouse ESCs (mESCs), hESCs, and hiPSCs (collectively named pluripotent stem cells, PSCs) to analyze brain microenvironments through the use of embryoid body (EB)-derived cells from these cell sources. EB are cell aggregates in 3D culture conditions that recapitulate embryonic development. Our approach focuses on studying the midbrain dopaminergic phenotype and transplanting EB into the substantia nigra pars compacta (SNpc) in a Parkinson's disease rodent model. Here, we describe cell culture protocols for EB generation from PSCs that show significant in vivo differentiation toward dopaminergic neurons., (© 2021. Springer Science+Business Media, LLC.)

Published
2022
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22. Management and outcome of obstructive ureteral stones in the emergency department: Emphasis on urine tests and antibiotics usage.

Author

Jennings CA, Khan Z, Sidhu P, Navarrete O, Palladino A, Rutland E, Villa JAC, Gonzalez LJ, Kalantari H, and Hassen GW

Subjects
Adult, Aged, Bacteriuria diagnosis, Bacteriuria etiology, Bacteriuria urine, Female, Follow-Up Studies, Humans, Male, Middle Aged, New York City, Pyuria diagnosis, Pyuria etiology, Pyuria urine, Retrospective Studies, Tomography, X-Ray Computed, Ureteral Calculi complications, Ureteral Calculi diagnosis, Ureteral Calculi urine, Urinary Tract Infections diagnosis, Urinary Tract Infections etiology, Urinary Tract Infections urine, Anti-Bacterial Agents therapeutic use, Bacteriuria drug therapy, Emergency Service, Hospital, Practice Patterns, Physicians' statistics & numerical data, Pyuria drug therapy, Ureteral Calculi therapy, Urinary Tract Infections drug therapy
Abstract

Background: Kidney stone related complaints in the Emergency Department (ED) are common. Current guidelines recommend antibiotic therapy for infected obstructive stones and stone removal in a timely fashion, but there is no clear recommendation for prophylactic antibiotic use for bacteriuria or pyuria in the setting of obstructive ureteral stones., Objectives: The aim of this study is to evaluate the current management of patients with obstructive ureteral stones in a single ED with emphasis on urine tests and antibiotics use., Methods: The picture archiving and communication system (PACS) was used to filter the list of patients who received a computed tomography (CT) scan of the abdomen and pelvis that positively identified obstructive ureteral stones. Demographics and clinical data were also recorded and analyzed., Results: Of the patients discharged, 278 patients did not receive antibiotics in the ED or a prescription. Of these, 8 patients had positive culture, 4 patients followed up, and one developed and was treated for a urinary-tract infection. One hundred ninety two patients were not given antibiotics in the ED but received an antibiotics prescription, and 4 patients had positive cultures grow. Two followed up and had no infection-related complications. Fourteen patients were discharged without a prescription after receiving a single dose of antibiotics in the ED, with no positive urine cultures and 9 patients following up without complication., Conclusion: Antibiotics were given at the discretion of the provider without clear pattern. A high rate of infectious complication did not occur in the followed up patient group., (Copyright © 2018. Published by Elsevier Inc.)

Published
2019
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23. Transcriptomic analysis reveals new hippocampal gene networks induced by prolactin.

Author

Cabrera-Reyes EA, Vanoye-Carlo A, Rodríguez-Dorantes M, Vázquez-Martínez ER, Rivero-Segura NA, Collazo-Navarrete O, and Cerbón M

Subjects
Animals, Female, Gene Expression Profiling methods, Lactation drug effects, Microglia drug effects, Neurogenesis drug effects, Neurogenesis genetics, Neurons drug effects, Neuroprotection drug effects, Neuroprotection genetics, Neuroprotective Agents pharmacology, Rats, Rats, Wistar, Gene Regulatory Networks drug effects, Gene Regulatory Networks genetics, Hippocampus drug effects, Prolactin pharmacology, Transcriptome drug effects, Transcriptome genetics
Abstract

Prolactin (Prl) is a pleiotropic hormone with multiple functions in several tissues and organs, including the brain. In the hippocampus, Prl has been implicated in several functions, including neuroprotection against excitotoxicity in lactating rats and in Prl-treated ovariectomized animals. However, the molecular mechanisms involved in Prl actions in the hippocampus have not been completely elucidated. The aim of this study was to analyse the hippocampal transcriptome of female Prl-treated ovariectomized rats. Transcriptomic analysis by RNASeq revealed 162 differentially expressed genes throughout 24 h of Prl treatment. Gene Ontology analysis of those genes showed that 37.65% were involved in brain processes that are regulated by the hippocampus, such as learning, memory and behaviour, as well as new processes that we did not foresee, such as glial differentiation, axogenesis, synaptic transmission, postsynaptic potential, and neuronal and glial migration. Immunodetection analysis demonstrated that Prl significantly modified microglial morphology, reduced the expression of Cd11b/c protein, and altered the content and location of the neuronal proteins Tau, Map2 and Syp, which are involved in axogenic and synaptic functions. This novel delineation of Prl activity in the hippocampus highlights its importance as a neuroactive hormone, opens a new avenue for understanding its actions and supports its participation in neuronal plasticity of this brain area.

Published
2019
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24. The Substantia Nigra Is Permissive and Gains Inductive Signals When Lesioned for Dopaminergic Differentiation of Embryonic Stem Cells.

Author

Collazo-Navarrete O, Hernández-García D, Guerrero-Flores G, Drucker-Colín R, Guerra-Crespo M, and Covarrubias L

Subjects
Animals, Cells, Cultured, Corpus Striatum cytology, Corpus Striatum metabolism, Dopaminergic Neurons cytology, Dopaminergic Neurons metabolism, Doublecortin Protein, Embryonic Stem Cells metabolism, Mice, Neural Stem Cells cytology, Neural Stem Cells metabolism, Neurogenesis physiology, Parkinson Disease metabolism, Substantia Nigra metabolism, Transcription Factors metabolism, Cell Differentiation physiology, Dopamine metabolism, Embryonic Stem Cells cytology, Substantia Nigra cytology
Abstract

Transplantation of dopaminergic (DA) cells into the striatum can rescue from dopamine deficiency in a Parkinson's disease condition, but this is not a suitable procedure for regaining the full control of motor activity. The minimal condition toward recovering the nigrostriatal pathway is the proper innervation of transplanted DA neurons or their precursors from the substancia nigra pars compacta (SNpc) to their target areas. However, functional integration of transplanted cells would require first that the host SNpc is suitable for their survival and/or differentiation. We recently reported that the intact adult SNpc holds a strong neurogenic environment, but primed embryonic stem cells (ie, embryoid body cells, EBCs) could not derive into DA neurons. In this study, we transplanted into the intact or lesioned SNpc, EBCs derived from embryonic stem cells that were prompt to differentiate into DA neurons by the forced expression of Lmx1a in neural precursor cells (R1B5/NesE-Lmx1a). We observed that, 6 days posttransplantation (dpt), R1B5 or R1B5/NesE-Lmx1a EBCs gave rise to Nes + and Dcx + cells within the host SNpc, but a large number of Th + cells derived only from EBCs exogenously expressing Lmx1a . In contrast, when transplantation was carried out into the 6-hydroxidopamine-lesioned SNpc, the emergence of Th + cells from EBCs was independent of exogenous Lmx1a expression, although these cells were not found by 15 dpt. These results suggest that the adult SNpc is not only a permissive niche for initiation of DA differentiation of non-neuralized cells but also releases factors upon damage that promote the acquisition of DA characteristics by transplanted EBCs.

Published
2019
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25. Functional determination of the differentiation potential of ventral mesencephalic neural precursor cells during dopaminergic neurogenesis.

Author

Guerrero-Flores G, Bastidas-Ponce A, Collazo-Navarrete O, Guerra-Crespo M, and Covarrubias L

Subjects
Animals, Astrocytes cytology, Cell Lineage, Cell Proliferation, Cell Survival, Dopaminergic Neurons metabolism, Embryo, Mammalian cytology, Female, Hedgehog Proteins metabolism, LIM-Homeodomain Proteins metabolism, Male, Mice, Transgenic, Models, Biological, Neural Stem Cells metabolism, Rats, Wistar, Stem Cell Niche, Transcription Factors metabolism, Cell Differentiation, Dopaminergic Neurons cytology, Mesencephalon cytology, Neural Stem Cells cytology, Neurogenesis
Abstract

The ventral mesencephalic neural precursor cells (vmNPCs) that give rise to dopaminergic (DA) neurons have been identified by the expression of distinct genes (e.g., Lmx1a, Foxa2, Msx1/2). However, the commitment of these NPCs to the mesencephalic DA neuronal fate has not been functionally determined. Evaluation of the plasticity of vmNPCs suggests that their commitment occurs after E10.5. Here we show that E9.5 vmNPCs implanted in an ectopic area of E10.5 mesencephalic explants, retained their specification marker Lmx1a and efficiently differentiated into neurons but did not express the gene encoding tyrosine hydroxylase (Th), the limiting enzyme for dopamine synthesis. A proportion of E10.5-E11.5 implanted vmNPCs behaved as committed, deriving into Th + neurons in ectopic sites. Interestingly, implanted cells from E12.5 embryos were unable to give rise to a significant number of Th + neurons. Concomitantly, differentiation assays in culture and in mesencephalic explants treated with Fgf2+LIF detected vmNPCs with astrogenic potential since E11.5. Despite this, a full suspension of E12.5 vmNPCs give rise to DA neurons in a similar proportion as those of E10.5 when they were transplanted into adult brain, but astrocytes were only detected with the former population. These data suggest that the subventricular postmitotic progenitors present in E12.5 ventral mesencephalon are unable to implant in embryonic explants and are the source of DA neurons in the transplanted adult brain. Based on our findings we propose that during DA differentiation committed vmNPCs emerge at E10.5 and they exhaust their neurogenic capacity with the rise of NPCs with astrogenic potential., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Published
2017
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26. Can the tandem measurement of age adjusted D-dimer and tissue plasminagen activator improve the clinical utility of a conventional D-dimer in the pulmonary embolism diagnosis?

Author

Flores J, García-Avello A, Ruíz A, Alonso E, Álvarez C, Navarrete O, and Arribas I

Subjects
Age Factors, Female, Hematologic Tests methods, Humans, Male, Middle Aged, Retrospective Studies, Fibrin Fibrinogen Degradation Products analysis, Pulmonary Embolism blood, Pulmonary Embolism diagnosis, Tissue Plasminogen Activator blood
Abstract

Background: The aim of this study was to evaluate if a sequential measurement of age adjust D-dimer (ADD) and tissue plasminogen activator (tPA) could increase the clinical utility in patients with suspected pulmonary embolism (PE) compared to a conventional D-dimer., Methods: We measured a conventional D-dimer (CDD), an ADD alone and a sequential combination ADD and tPA (ADD/tPA combination) in a prospective sample of 127 outpatients with PE suspected. Diagnosis of PE was based on a strict protocol. Plasma sample to measure levels of tPA and D-dimer was obtained at enrollment, and CDD, ADD and tPA were assessed at the end of study. For CDD the cut-off value was 500 ng/mL and for ADD the cut-off value was defined as (patient's age x10) ng/mL in patients aged >50. We compared the sensitivity, specificity and clinical utility obtained for CDD, ADD alone, and ADD/tPA combination., Results: PE was confirmed in 41 patients (32%). The sensitivity, specificity and clinical utility for CDD were 95%, 36% and 28%, respectively. The ADD/tPA combination and ADD alone demonstrated an increased in specificity of +29% and +12% respectively, and increased in clinical utility of +20% and +8%, respectively, compared to CDD, and this was obtained without loss of sensitivity., Conclusion: The ADD/tPA combination substantially increased the clinical utility in the PE diagnosis compared with conventional D-dimer, without reducing the security. The ADD/tPA combination could decrease the need for pulmonary vascular imaging for the PE diagnosis in nearly the half. These promising results should be validated prospectively.

Published
2016

27. Mouse embryonic stem cell-derived cells reveal niches that support neuronal differentiation in the adult rat brain.

Author

Maya-Espinosa G, Collazo-Navarrete O, Millán-Aldaco D, Palomero-Rivero M, Guerrero-Flores G, Drucker-Colín R, Covarrubias L, and Guerra-Crespo M

Subjects
Animals, Brain Ischemia metabolism, Brain Ischemia pathology, Brain Ischemia therapy, Cell Line, Corpus Striatum pathology, Embryonic Stem Cells pathology, GABAergic Neurons pathology, Heterografts, Male, Mice, Rats, Rats, Wistar, Stroke metabolism, Stroke pathology, Stroke therapy, Cell Differentiation, Corpus Striatum metabolism, Embryonic Stem Cells metabolism, GABAergic Neurons metabolism, Stem Cell Niche, Stem Cell Transplantation
Abstract

A neurogenic niche can be identified by the proliferation and differentiation of its naturally residing neural stem cells. However, it remains unclear whether "silent" neurogenic niches or regions suitable for neural differentiation, other than the areas of active neurogenesis, exist in the adult brain. Embryoid body (EB) cells derived from embryonic stem cells (ESCs) are endowed with a high potential to respond to specification and neuralization signals of the embryo. Hence, to identify microenvironments in the postnatal and adult rat brain with the capacity to support neuronal differentiation, we transplanted dissociated EB cells to conventional neurogenic and non-neurogenic regions. Our results show a neuronal differentiation pattern of EB cells that was dependent on the host region. Efficient neuronal differentiation of EB cells occurred within an adjacent region to the rostral migratory stream. EB cell differentiation was initially patchy and progressed toward an even distribution along the graft by 15-21 days post-transplantation, giving rise mostly to GABAergic neurons. EB cells in the striatum displayed a lower level of neuronal differentiation and derived into a significant number of astrocytes. Remarkably, when EB cells were transplanted to the striatum of adult rats after a local ischemic stroke, increased number of neuroblasts and neurons were observed. Unexpectedly, we determined that the adult substantia nigra pars compacta, considered a non-neurogenic area, harbors a robust neurogenic environment. Therefore, neurally uncommitted cells derived from ESCs can detect regions that support neuronal differentiation within the adult brain, a fundamental step for the development of stem cell-based replacement therapies., (© 2014 AlphaMed Press.)

Published
2015
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