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103. Comparative pangenomic analysis of Campylobacter fetus isolated from Spanish bulls and other mammalian species

Author

Pena‑Fernández, Nerea, Ocejo, Medelin, Van der Graaf‑van Bloois, Linda, Lavín, José Luis, Kortabarria, Nekane, Collantes Fernández, Esther, Hurtado, Ana, Aduriz, Gorka, Pena‑Fernández, Nerea, Ocejo, Medelin, Van der Graaf‑van Bloois, Linda, Lavín, José Luis, Kortabarria, Nekane, Collantes Fernández, Esther, Hurtado, Ana, and Aduriz, Gorka

Abstract

Author contributions: G.A. and A.H. conceived the study and participated in its design. N.P.F. and N.K. performed laboratory analysis. N.P.F., M.O., J.L.L. and L.G. participated in the bioinformatics analysis. N.P.F. wrote the manuscript, with interpretation of results and discussion inputs from GA, AH, E.C., M.O., N.K., J.L.L. and L.G. All authors read and approved the fnal manuscript., Campylobacter fetus comprises two closely related mammal-associated subspecies: Campylobacter fetus subsp. fetus (Cff) and Campylobacter fetus subsp. venerealis (Cfv). The latter causes bovine genital campylobacteriosis, a sexually-transmitted disease endemic in Spain that results in significant economic losses in the cattle industry. Here, 33 C. fetus Spanish isolates were whole-genome sequenced and compared with 62 publicly available C. fetus genomes from other countries. Genome-based taxonomic identification revealed high concordance with in silico PCR, confirming Spanish isolates as Cff (n = 4), Cfv (n = 9) and Cfv biovar intermedius (Cfvi, n = 20). MLST analysis assigned the Spanish isolates to 6 STs, including three novel: ST-76 and ST-77 for Cfv and ST-78 for Cff. Core genome SNP phylogenetic analysis of the 95 genomes identified multiple clusters, revealing associations at subspecies and biovar level between genomes with the same ST and separating the Cfvi genomes from Spain and other countries. A genome-wide association study identified pqqL as a Cfv-specific gene and a potential candidate for more accurate identification methods. Functionality analysis revealed variations in the accessory genome of C. fetus subspecies and biovars that deserve further studies. These results provide valuable information about the regional variants of C. fetus present in Spain and the genetic diversity and predicted functionality of the different subspecies., Ministerio de Ciencia e Innovación (España), Depto. de Sanidad Animal, Fac. de Veterinaria, TRUE, pub

Published
2024

104. Adaptación climática en Maputo. Viviendas resilientes

Author

Moreno Fernández, Esther, Domínguez Mateos, Esther, Moreno Fernández, Esther, and Domínguez Mateos, Esther

Abstract

Mozambique, donde tiene lugar este trabajo es, en la actualidad, uno de los países más pobres y con menos recursos en el mundo. Más de la mitad de su población vive por debajo del umbral de la pobreza y su economía depende fuertemente de la ayuda exterior. Debido a esto, la atención de la población se centra en solucionar problemas de primera necesidad como la alimentación y la vivienda. Por otro lado, existe a nivel global, una gran preocupación para lograr disminuir los efectos provocados por el cambio climático. Esta cuestión es bastante discutida en los países más privilegiados, debido al rechazo por parte de la población a renunciar a determinadas comodidades y privilegios que poseen pero, ¿quién se preocupa de cómo afectará a los países con menos recursos y carentes de los derechos básicos? Es necesario y urgente implementar acciones para lograr minimizar las consecuencias del calentamiento global sobre los países en vías de desarrollo, debido a que empeorará la situación de vulnerabilidad en la que se encuentran actualmente. El estudio realizado en este Trabajo de Fin de Grado pretende contribuir a mejorar la precaria situación en la que se encuentra la vivienda en Maputo, capital de Mozambique. Tras un estudio de la evolución de las viviendas que han existido en la zona, se ha elaborado una guía de autoconstrucción de casas accesibles fabricadas con materiales locales y que, a su vez estén acondicionadas para resistir los efectos del cambio climático (Figura 1). De esta manera, se cubre el derecho básico de acceso a una vivienda y se facilita la habitabilidad y el mantenimiento de estas edificaciones.

Published
2024

105. Resultados y características del trasplante autólogo de progenitores en Linfoma No Hodgkin B difuso de células grandes en recidiva o refractariedad

Author

Nieto Cantalapiedra, María del Pilar, Fernández Fernández, Esther, Universidad de Valladolid. Facultad de Medicina, Nieto Cantalapiedra, María del Pilar, Fernández Fernández, Esther, and Universidad de Valladolid. Facultad de Medicina

Abstract

El linfoma difuso de células B grandes es el Linfoma No Hodgkin más frecuente y agresivo en adultos. Con la terapia de primera línea con R-CHOP se consigue una supervivencia a largo plazo de 60-70%. Un 30-40% de los pacientes con LDCBG recaen o son refractarios al tratamiento de primera línea. Se les valora para trasplante autólogo de progenitores hematopoyéticos (TAPH) y en caso de ser aptos se administra quimioterapia de rescate y en aquellos casos quimiosensibles se realiza quimioterapia mieloablativa seguida del TAPH. Aproximadamente el 50% de los pacientes responden a la quimioterapia de rescate y por lo tanto pueden someterse al TAPH. El TAPH presenta una tasa de curación global del 25-30%. Para aquellos pacientes no candidatos al TAPH o en recaída o refractariedad tras el TAPH, hasta hace poco han sido tratados con intenciones paliativas. La aparición de nuevas terapias como la CAR-T, según los estudios publicados, parece ser una alternativa eficaz y segura para estos pacientes. En este estudio retrospectivo, se han seleccionado 33 pacientes con LBDCG R/R tratados mediante TAPH entre los años 2007 y 2022 en el HURH y en el HCUV, para estudiar la supervivencia global y la supervivencia libre de progresión tras el TAPH mediante curvas de supervivencia de Kaplan-Meier y hacer una comparación entre los pacientes con recaída precoz (POD12) vs tardía (no POD12). Los resultados obtenidos muestran unos resultados favorables, con supervivencias globales y libres de progresión a largo plazo superiores al 70%. Por otra parte, los pacientes POD12 presentan peores resultados frente a los pacientes no POD12, aunque de manera no significativa. En definitiva, los resultados señalan que el TAPH sigue siendo una opción eficaz y segura frente a las nuevas terapias., Grado en Medicina

Published
2024

106. Comparative pangenomic analysis of Campylobacter fetus isolated from Spanish bulls and other mammalian species

Author

Klinische infectiologie en microb. lab., Infectious Diseases and Immunology - KLIF, Pena-Fernández, Nerea, Ocejo, Medelin, van der Graaf-van Bloois, Linda, Lavín, Jose Luís, Kortabarria, Nekane, Collantes-Fernández, Esther, Hurtado, Ana, Aduriz, Gorka, Klinische infectiologie en microb. lab., Infectious Diseases and Immunology - KLIF, Pena-Fernández, Nerea, Ocejo, Medelin, van der Graaf-van Bloois, Linda, Lavín, Jose Luís, Kortabarria, Nekane, Collantes-Fernández, Esther, Hurtado, Ana, and Aduriz, Gorka

Published
2024

107. Pancreatic adenocarcinoma treated with surgical resection, toceranib phosphate and firocoxib in a dog: a case report

Author

Fuertes Recuero, Manuel, Martínez De Merlo, Elena, Lizasoain Sanz, Guillermo, Arroba Alonso, Amanda, Sánchez López, Alejandro, Suárez Redondo, María, Ortiz Díez, Gustavo, Vázquez Fernández, Esther, Fuertes Recuero, Manuel, Martínez De Merlo, Elena, Lizasoain Sanz, Guillermo, Arroba Alonso, Amanda, Sánchez López, Alejandro, Suárez Redondo, María, Ortiz Díez, Gustavo, and Vázquez Fernández, Esther

Abstract

Author contributions: All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by MFR, GLS, AAA, ASL, EMM, MSR, EVF and GOD. The first draft of the manuscript was written by MFR and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript., Exocrine pancreatic carcinomas are rarely reported in dogs. A ductal pancreatic adenocarcinoma in a 10-year-old intact beagle is described in this report. The diagnosis was made based on clinical signs, imaging (abdominal ultrasound and CT scan) and histopathology. Treatment consisted of partial right lobe pancreatectomy followed by adjuvant therapy with toceranib phosphate (Palladia®) and firocoxib (Previcox®) for six months. The treatment was well tolerated, and the survival time was 445 days. To our knowledge, this is the longest survival reported in the literature for a dog diagnosed with exocrine pancreatic adenocarcinoma. The results described here may contribute to provide a better understanding about this neoplasia and potential treatment options., Consejo Superior de Investigaciones Científicas, Depto. de Sanidad Animal, Centro de Vigilancia Sanitaria Veterinaria (VISAVET), Fac. de Veterinaria, TRUE, pub

Published
2024

108. In vivo and in vitro models show unexpected degrees of virulence among Toxoplasma gondii type II and III isolates from sheep

Author

Fernández Escobar, Mercedes, Ortega Mora, Luis Miguel, Collantes Fernández, Esther, Calero Bernal, Rafael, Regidor-Cerrillo, Javier, Vallejo, Raquel, Benavides, Julio, Fernández Escobar, Mercedes, Ortega Mora, Luis Miguel, Collantes Fernández, Esther, Calero Bernal, Rafael, Regidor-Cerrillo, Javier, Vallejo, Raquel, and Benavides, Julio

Abstract

Author's Contributions MF-E, RC-B, JR-C, EC-F, and LO-M conceived and designed the laboratory tests. MF-E, RC-B, RV, and JB performed experiments. MF-E, RC-B, RV, JR-C, and EC-F analysed the data. LO-M, EC-F, and JB contributed reagents/materials/analysis tools. MF-E, RC-B, JR-C, JB, EC-F, and LO-M drafted the manuscript. All authors read and approved the final manuscript., Toxoplasma gondii is an important zoonotic agent with high genetic diversity, complex epidemiology, and variable clinical outcomes in animals and humans. In veterinary medicine, this apicomplexan parasite is considered one of the main infectious agents responsible for reproductive failure in small ruminants worldwide. The aim of this study was to phenotypically characterize 10 Spanish T. gondii isolates recently obtained from sheep in a normalized mouse model and in an ovine trophoblast cell line (AH-1) as infection target cells. The panel of isolates met selection criteria regarding such parameters as genetic diversity [types II (ToxoDB #1 and #3) and III (#2)], geographical location, and sample of origin (aborted foetal brain tissues or adult sheep myocardium). Evaluations of in vivo mortality, morbidity, parasite burden and histopathology were performed. Important variations between isolates were observed, although all isolates were classified as “nonvirulent” (< 30% cumulative mortality). The isolates TgShSp16 (#3) and TgShSp24 (#2) presented higher degrees of virulence. Significant differences were found in terms of in vitro invasion rates and tachyzoite yield at 72 h post-inoculation (hpi) between TgShSp1 and TgShSp24 isolates, which exhibited the lowest and highest rates, respectively. The study of the CS3, ROP18 and ROP5 loci allelic profiles revealed only type III alleles in ToxoDB #2 isolates and type II alleles in the #1 and #3 isolates included. We concluded that there are relevant intra- and inter-genotype virulence differences in Spanish T. gondii isolates, which could not be inferred by genetic characterization using currently described molecular markers., Ministerio de Ciencia e Innovación (AGL2016-75935-C2-R), Comunidad de Madrid (PLATESA2-CMP2018/BAA-4370), Depto. de Sanidad Animal, Fac. de Veterinaria, TRUE, pub

Published
2024

109. Prevalence of Bovine Genital Campylobacteriosis, Associated Risk Factors and Spatial Distribution in Spanish Beef Cattle Based on Veterinary Laboratory Database Records

Author

Nerea Pena-Fernández, David Cano-Terriza, Ignacio García-Bocanegra, Horcajo Iglesias, María Del Pilar, Vázquez Arbáizar, María Patricia, Darío Cleofé-Resta, Pérez Arroyo, Bárbara, Ortega Mora, Luis Miguel, Collantes Fernández, Esther, Nerea Pena-Fernández, David Cano-Terriza, Ignacio García-Bocanegra, Horcajo Iglesias, María Del Pilar, Vázquez Arbáizar, María Patricia, Darío Cleofé-Resta, Pérez Arroyo, Bárbara, Ortega Mora, Luis Miguel, and Collantes Fernández, Esther

Abstract

Bovine genital campylobacteriosis (BGC) is a sexually transmitted disease that causes early reproductive failure in natural breeding cattle that are managed extensively. The aim of this study was to assess the BGC prevalence in Spain from 2011 to 2019 using data collected cross-sectionally from the diagnostic reports issued by the SALUVET veterinary diagnostic laboratory from a total of 5,182 breeding bulls from 1,950 herds managed under "dehesa" systems (large herds within fenced pastures and all-year breeding season) or mountain systems (smaller herds with seasonal breeding management and grazing in communal mountain pastures). Infection was detected by PCR in 7.7 and 12.2% of the bulls and herds tested, respectively. The "dehesa" herd management system (OR = 2.078, P = < 0.001, 95% CI = 1.55-1.77), bovine trichomonosis status of the herd (OR = 1.606, P = 0.004, 95% CI = 1.15-2.22), and bulls ≥3 years old (OR = 1.392, P = 0.04, 95% CI = 1.01-1.92) were identified as risk factors associated with Campylobacter fetus venerealis infection. We also studied the high-risk areas for circulation of the infection in extensive beef cattle herds in Spain, showing four significant clusters in "dehesa" areas in the south-western provinces of the country and a fifth cluster located in a mountain area in northern Spain. The results obtained in the present study indicate that BGC is endemic and widely distributed in Spanish beef herds. Specifically, "dehesa" herds are at greater risk for introduction of Cfv based on relatively high local prevalence of the infection and the use of specific management practices., Depto. de Sanidad Animal, Fac. de Veterinaria, TRUE, pub

Published
2024

110. Prevalence of intestinal parasite infections in stray and farm dogs from Spain

Author

Javier Regidor-Cerrillo, Arranz-Solís, David, Moreno Gonzalo, Javier, Pedraza-Díaz, Susana, Gómez Bautista, Mercedes, Ortega Mora, Luis Miguel, Collantes Fernández, Esther, Javier Regidor-Cerrillo, Arranz-Solís, David, Moreno Gonzalo, Javier, Pedraza-Díaz, Susana, Gómez Bautista, Mercedes, Ortega Mora, Luis Miguel, and Collantes Fernández, Esther

Abstract

Dogs play a potential role as reservoirs for zoonotic parasites, being especially problematic uncontrolled dog populations such as stray and farm dogs with access to populated areas. In order to investigate the prevalence of canine intestinal parasites in at-risk dog populations, we tested a total of 233 faecal samples shed by stray and dairy farm dogs from northern Spain. Telemann method was used to detect the presence of eggs and (oo)cysts of common dog intestinal parasites and Cryptosporidium was detected by PCR. One hundred and forty eight out of 233 samples (63.5%) were positive for at least one intestinal parasite, being Ancylostomidae (35.6%; 83/233) and Trichuris (35.2%; 82/233) the parasites most frequently identified. Cryptosporidium DNA was not detected in any of the faecal samples analysed. The overall prevalence was significantly higher in stray dogs than in farm dogs (72.5% vs 58.8%). Specifically, stray dogs had a significantly higher prevalence of Ancylostomatidae, Toxocara, Toxascaris and Taenidae. These dog populations are an important source of environmental contamination with intestinal parasite forms, which could be of significance to animal and human health., Universidad Complutense de Madrid (PR1/06-14467-B), Depto. de Sanidad Animal, Fac. de Veterinaria, TRUE, pub

Published
2024

115. TaqMan-quantitative PCR assays applied in Neospora caninum knock-outs generated through CRISPR-Cas9 allow to determine the copy numbers of integrated dihydrofolate reductasethymidylate synthase drug selectable markers.

Author

Rico-San Román, Laura, Alexander Hänggeli, Kai Pascal, Hemphill, Andrew, Horcajo, Pilar, Collantes-Fernández, Esther, Miguel Ortega-Mora, Luis, and Boubaker, Ghalia

Subjects
NEOSPORA caninum, NEUROMUSCULAR diseases, CATTLE diseases, DOG diseases, DRUG target, GENOME editing, CRISPRS
Abstract

As for many other organisms, CRISPR-Cas9 mediated genetic modification has gained increasing importance for the identification of vaccine candidates and drug targets in Neospora caninum, an apicomplexan parasite causing abortion in cattle and neuromuscular disease in dogs. A widely used approach for generating knock-out (KO) strains devoid of virulence factors is the integration of a drug selectable marker such as mutated dihydrofolate reductase-thymidylate synthase (mdhfr-ts) into the target gene, thus preventing the synthesis of respective protein and mediating resistance to pyrimethamine. However, CRISPR-Cas9 mutagenesis is not free of off-target effects, which can lead to integration of multiple mdhfr-ts copies into other sites of the genome. To determine the number of integrated mdhfr-ts in N. caninum, a duplex quantitative TaqMan PCR was developed. For this purpose, primers were designed that amplifies a 106 bp fragment from wild-type (WT) parasites corresponding to the single copy wtdhfrs-ts gene, as well as the mutated mdhfrs-ts present in KO parasites that confers resistance and were used simultaneously with primers amplifying the diagnostic NC5 gene. Thus, the dhfr-ts to NC5 ratio should be approximately 1 in WT parasites, while in KO parasites with a single integrated mdhrf-ts gene this ratio is doubled, and in case of multiple integration events even higher. This approach was applied to the Neospora KO strains NcDGRA7 and NcDROP40. For NcDGRA7, the number of tachyzoites determined by dhfr-ts quantification was twice the number of tachyzoites determined by NC5 quantification, thus indicating that only one mdhfr-ts copy was integrated. The results obtained with the NcDROP40 strain, however, showed that the number of dhfr-ts copies per genome was substantially higher, indicating that at least three copies of the selectable mdhfrts marker were integrated into the genomic DNA during gene editing by CRISPRCas9. This duplex TaqMan-qPCR provides a reliable and easy-to-use tool for assessing CRISPR-Cas9 mediated mutagenesis in WT N. caninum strains. [ABSTRACT FROM AUTHOR]

Published
2024
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125. Trichomonas

Author

Collántes-Fernández, Esther, Fort, Marcelo C., Ortega-Mora, Luis M., Schares, Gereon, Florin-Christensen, Monica, editor, and Schnittger, Leonhard, editor

Published
2018
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126. Factors associated with poor anticoagulation control with vitamin K antagonists among outpatients attended in Internal Medicine and Neurology. The ALADIN study

Author

Pose, Antonio, Díaz, José Antonio, Rodríguez, Manuel, Pena, Marta, Arias, Susana, Larrosa, Davinia, González, Álvaro, Rodríguez, Elisa, González, Montserrat, Fernández, Dolores, Barbagelata, Cristina, Raña, Natividad, Freire, Santiago, Cerqueiro, J. Manuel, Guerrero, Héctor, Ramos, Laura, Álvarez, Leticia, de Lis, José M., Caro, Carmen, Seijo, Manuel, Mederer, Susana, de Zarraga, Miguel Alberto, Ferreiro, José, Terrero, José M., Arias, Mónica, Pérez, Roberto, Sánchez, Joaquín, Maciñeiras, José, Fernández, Julián, Jaén, Fernando, Esteva, David, Zamora, Mónica, Navarrete, Nuria, García, Javier, Mérida, Luis, Corrales, Miguel Ángel, Quirós, Raúl, Cantero, Jesús, Barrero, Francisco Javier, Villegas, Inmaculada, Castro, José, Foronda, Jesús, Carrillo, Dionisio, Vega, Jesús, Trujillo, José Antonio, Montero, Manuel, Jurado, Ana, Sánchez, Carlos, Agüera-Morales, Eduardo, Sánchez, María, Durán, Purificación, Fernández de la Puerta, Rafael, Pérez de la Blanca, María, Martínez, María Paz, Fernández, Óscar, Tamayo, José Antonio, Bustamante, Rafael, Serrano, Pedro Jesús, Arjona, Antonio, Fernández, Javier, Payan, Manuel, Gómez, Ricardo, Peña, Daniel, Cabrerizo, Enrique, Salgado, Fernando, Ivanova Georgieva, Radka, Gil-Núñez, Antonio, Bello, Elena, Díaz, Fernando, Medina, Antonio, Castellano, Ana, Miranda, Yesica, Fabre, Óscar, García Polo, Iluminada, Ibáñez, Patricia, Sainz, Clara, Sierra, Fernando, Aragón, Esther, Díaz, Jaime, Aguilar, Fernando, Ortega, María Ángeles, Egido, José Antonio, Pontes, José Carlos, García, Miguel Ángel, Cabrera, Francisco, Batalla, Blanca, Culla, Alex, Molina, Carlos, Flores, Alan, Seró, Laia, Muchada, Marian, Meler, Pilar, Boned-Riera, Sandra, Cánovas, David, Estela, Jordi, Font, Juli, Purroy, Francisco, Benabdelhak, Ikram, Sanahuja, Jordi, Roquer, Jaume, Rodríguez, Ana, Ois, Ángel, Cuadrado, Elisa, Jiménez, Jordi, Nogués, Xavier, Kuprinski, Jerzy, German, Antoni, Irigoyen, Daniel, Cara, Juan José, Font, Maria Àngels, Huertas, Sonia, Martínez-Domeño, Alejandro, Arroyo, Juan Antonio, Delgado-Mederos, Raquel, Gómez-Choco, Manuel Jesús, Mengual, Juan José, García, Sonia M., Castellanos, M. del Mar, van Eedenburg, Cecile, Cañas, Imma, Espinosa, Jordi, Montull, Santiago, Quesada, Helena, Ustrell, Xavier, Homedes, Christian, Navalpotro, Irene, Casanova, Jordi, Lago, Aida Pilar, Morata, Carmen, Gorriz, David, Moreno, Iván, Tembl, José, Ponz, Alejandro, Fonseca, M. José, Chamarro, Raquel, Gil, Rosario, Oliver, Vicente, Pampliega, Ana, Artero, Arturo, Puchades, Francesc, Landete, Lamberto, Vilar, Carlos, Jiménez, Carmen, Vives, Bárbara, Moragues, M. Dolores, Díaz, Rosa, Tur, Silvia, Escribano, José Bernardo, Lucas, César, Martínez, Francisco, Pons, José Miguel, Romero, Amparo, García, David, Pérez, José, Villaverde, Ramón, Martínez, Salvadora, Rodríguez, Aida, Tejero, Carlos, Pérez, Cristina, Mostacero, Enrique, Fernández, Covadonga, Luna, Alain, Pérez, Tomás, González, Félix, de Arce, Ana, Martínez, Maite, Díez, Noemí, Gállego, Jaime, Zandio, Beatriz, Herrera, María, Aymerich, Nuria, Muñoz, Roberto, Marta, Javier, Artal, Jorge, Errea, José M., Timiraos, Juan José, Moreno, M. Pilar, Freijo, Mar, García, Juan Manuel, Gil, M. Carmen, Revilla, M. Ángeles, Palacio, Enrique, Vázquez, José Luis, Bestué, María, Latorre, Ana, Calvo, Eva, Ballester, Laura, Serrano, Marta, Juega, Jesús M., López, M. Ángeles, Irimia, Pablo, Imaz, Laura, Fuentes, Blanca, Sanz, Borja Enrique, Beltrán, Luis, Ruiz, Gerardo, Martínez, Patricia, Sánchez, Demetrio, Barroso, Emilio, Molina, Igor, Budiño, Marco Antonio, Masjuan, Jaime, de Felipe, Alicia, Matute, Consuelo, Tejada, Javier, Morán, Alberto, Fernández, Esther, Riveira, M. del Carmen, Carnedo, Joaquín, Manquillo, Antonio, González, Raquel, Fernández, José Carlos, Guillan, Marta, Yebra, Miguel, Trejo, José M., Saiz, Jesús, Martínez-Acitores, Juan Carlos, Bravo, Yolanda, Arenillas, Juan Francisco, Calleja, Ana, Cortijo, Elisa, Reyes, Javier, López, Luis, Muñoz, Pedro Luis, Fidalgo, M. Ángeles, Hernández, Jacinto, Gómez, José Carlos, Morán, José Carlos, Gonzalo, Sonia, Marrero, Jorge, Satué, José Ángel, Belinchón, Juan Carlos, Moniche, Francisco, Calderón, Enrique, Escudero, Irene, de la Torre, Javier, Casado, Ignacio, Antón, Joaquín, Portilla, Juan Carlos, Luengo, Juan, Rosal, Joan, Calzado, Elena, Anglada, Juan Carlos, Girón, Juan, Ramírez, José M., Pijierro, Agustín, Roa, Ana, Romero, Jorge, Aguayo, Mariano, Borrachero, Cristina, Sanz, Gema, Gómez, M. José, Rico, Miguel Ángel, Cayon, Antonio, Carmona, Eduardo, Cerro, Román, López, Rafael, Aguirre, Alfonso, Lozano, Francisco, Rivera, José M., Contreras Muruaga, M.M., Reig, G., Vivancos, J., González, A., Cardona, P., Ramírez-Moreno, J.M., Martí-Fábregas, J., and Suárez Fernández, C.

Published
2018
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127. Factores asociados al mal control de la anticoagulación con antivitamina K en pacientes con fibrilación auricular no valvular atendidos en consultas de Medicina Interna y Neurología. Estudio ALADIN

Author

Pose, Antonio, Díaz, José Antonio, Rodríguez, Manuel, Pena, Marta, Arias, Susana, Larrosa, Davinia, González, Álvaro, Rodríguez, Elisa, González, Montserrat, Fernández, Dolores, Barbagelata, Cristina, Raña, Natividad, Freire, Santiago, Cerqueiro, J. Manuel, Guerrero, Héctor, Ramos, Laura, Álvarez, Leticia, de Lis, José M., Caro, Carmen, Seijo, Manuel, Mederer, Susana, de Zarraga, Miguel Alberto, Ferreiro, José, Terrero, José M., Arias, Mónica, Pérez, Roberto, Sánchez, Joaquín, Maciñeiras, José, Fernández, Julián, Jaén, Fernando, Esteva, David, Zamora, Mónica, Navarrete, Nuria, García, Javier, Mérida, Luis, Corrales, Miguel Ángel, Quirós, Raúl, Cantero, Jesús, Barrero, Francisco Javier, Villegas, Inmaculada, Castro, José, Foronda, Jesús, Carrillo, Dionisio, Vega, Jesús, Trujillo, José Antonio, Montero, Manuel, Jurado, Ana, Sánchez, Carlos, Agüera-Morales, Eduardo, Sánchez, María, Durán, Purificación, Fernández de la Puerta, Rafael, Pérez de la Blanca, María, Martínez, María Paz, Fernández, Óscar, Tamayo, José Antonio, Bustamante, Rafael, Serrano, Pedro Jesús, Arjona, Antonio, Fernández, Javier, Payan, Manuel, Gómez, Ricardo, Peña, Daniel, Cabrerizo, Enrique, Salgado, Fernando, Ivanova Georgieva, Radka, Gil-Núñez, Antonio, Bello, Elena, Díaz, Fernando, Medina, Antonio, Castellano, Ana, Miranda, Yesica, Fabre, Óscar, García Polo, Iluminada, Ibáñez, Patricia, Sainz, Clara, Sierra, Fernando, Aragón, Esther, Díaz, Jaime, Aguilar, Fernando, Ortega, María Ángeles, Egido, José Antonio, Pontes, José Carlos, García, Miguel Ángel, Cabrera, Francisco, Batalla, Blanca, Culla, Alex, Molina, Carlos, Flores, Alan, Seró, Laia, Muchada, Marian, Meler, Pilar, Boned-Riera, Sandra, Cánovas, David, Estela, Jordi, Font, Juli, Purroy, Francisco, Benabdelhak, Ikram, Sanahuja, Jordi, Roquer, Jaume, Rodríguez, Ana, Ois, Ángel, Cuadrado, Elisa, Jiménez, Jordi, Nogués, Xavier, Kuprinski, Jerzy, German, Antoni, Irigoyen, Daniel, Cara, Juan José, Font, Maria Àngels, Huertas, Sonia, Martínez-Domeño, Alejandro, Arroyo, Juan Antonio, Delgado-Mederos, Raquel, Gómez-Choco, Manuel Jesús, Mengual, Juan José, García, Sonia M., Castellanos, M. del Mar, van Eedenburg, Cecile, Cañas, Imma, Espinosa, Jordi, Montull, Santiago, Quesada, Helena, Ustrell, Xavier, Homedes, Christian, Navalpotro, Irene, Casanova, Jordi, Lago, Aida Pilar, Morata, Carmen, Gorriz, David, Moreno, Iván, Tembl, José, Ponz, Alejandro, Fonseca, M. José, Chamarro, Raquel, Gil, Rosario, Oliver, Vicente, Pampliega, Ana, Artero, Arturo, Puchades, Francesc, Landete, Lamberto, Vilar, Carlos, Jiménez, Carmen, Vives, Bárbara, Moragues, M. Dolores, Díaz, Rosa, Tur, Silvia, Escribano, José Bernardo, Lucas, César, Martínez, Francisco, Pons, José Miguel, Romero, Amparo, García, David, Pérez, José, Villaverde, Ramón, Martínez, Salvadora, Rodríguez, Aida, Tejero, Carlos, Pérez, Cristina, Mostacero, Enrique, Fernández, Covadonga, Luna, Alain, Pérez, Tomás, González, Félix, de Arce, Ana, Martínez, Maite, Díez, Noemí, Gállego, Jaime, Zandio, Beatriz, Herrera, María, Aymerich, Nuria, Muñoz, Roberto, Marta, Javier, Artal, Jorge, Errea, José M., Timiraos, Juan José, Moreno, M. Pilar, Freijo, Mar, García, Juan Manuel, Gil, M. Carmen, Revilla, M. Ángeles, Palacio, Enrique, Vázquez, José Luis, Bestué, María, Latorre, Ana, Calvo, Eva, Ballester, Laura, Serrano, Marta, Juega, Jesús M., López, M. Ángeles, Irimia, Pablo, Imaz, Laura, Fuentes, Blanca, Sanz, Borja Enrique, Beltrán, Luis, Ruiz, Gerardo, Martínez, Patricia, Sánchez, Demetrio, Barroso, Emilio, Molina, Igor, Budiño, Marco Antonio, Masjuan, Jaime, de Felipe, Alicia, Matute, Consuelo, Tejada, Javier, Morán, Alberto, Fernández, Esther, Riveira, M. del Carmen, Carnedo, Joaquín, Manquillo, Antonio, González, Raquel, Fernández, José Carlos, Guillan, Marta, Yebra, Miguel, Trejo, José M., Saiz, Jesús, Martínez-Acitores, Juan Carlos, Bravo, Yolanda, Arenillas, Juan Francisco, Calleja, Ana, Cortijo, Elisa, Reyes, Javier, López, Luis, Muñoz, Pedro Luis, Fidalgo, M. Ángeles, Hernández, Jacinto, Gómez, José Carlos, Morán, José Carlos, Gonzalo, Sonia, Marrero, Jorge, Satué, José Ángel, Belinchón, Juan Carlos, Moniche, Francisco, Calderón, Enrique, Escudero, Irene, de la Torre, Javier, Casado, Ignacio, Antón, Joaquín, Portilla, Juan Carlos, Luengo, Juan, Rosal, Joan, Calzado, Elena, Anglada, Juan Carlos, Girón, Juan, Ramírez, José M., Pijierro, Agustín, Roa, Ana, Romero, Jorge, Aguayo, Mariano, Borrachero, Cristina, Sanz, Gema, Gómez, M. José, Rico, Miguel Ángel, Cayon, Antonio, Carmona, Eduardo, Cerro, Román, López, Rafael, Aguirre, Alfonso, Lozano, Francisco, Rivera., José M., Contreras Muruaga, M.M., Reig, G., Vivancos, J., González, A., Cardona, P., Ramírez-Moreno, J.M., Martí-Fábregas, J., and Suárez Fernández, C.

Published
2018
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128. Cholesterol metabolism in Mycobacterium smegmatis

Author

García‐Fernández, Esther, Frank, Daniel J, Galán, Beatriz, Kells, Petrea M, Podust, Larissa M, García, José L, and de Montellano, Paul R Ortiz

Subjects
Biochemistry and Cell Biology, Biological Sciences, Alcoholism, Alcohol Use and Health, Rare Diseases, Emerging Infectious Diseases, Genetics, Substance Misuse, Infectious Diseases, Tuberculosis, Aetiology, 2.2 Factors relating to the physical environment, Infection, Good Health and Well Being, Antifungal Agents, Azoles, Binding Sites, Cholestenones, Cholesterol, Crystallography, X-Ray, Cytochrome P-450 Enzyme System, Gene Deletion, Gene Expression Regulation, Bacterial, Hydroxylation, Models, Molecular, Mycobacterium smegmatis, Mycobacterium tuberculosis, Oxidation-Reduction, Protein Structure, Tertiary, Evolutionary Biology, Microbiology, Ecology
Abstract

Degradation of the cholesterol side-chain in Mycobacterium tuberculosis is initiated by two cytochromes P450, CYP125A1 and CYP142A1, that sequentially oxidize C26 to the alcohol, aldehyde and acid metabolites. Here we report characterization of the homologous enzymes CYP125A3 and CYP142A2 from Mycobacterium smegmatis mc(2) 155. Heterologously expressed, purified CYP125A3 and CYP142A2 bound cholesterol, 4-cholesten-3-one, and antifungal azole drugs. CYP125A3 or CYP142A2 reconstituted with spinach ferredoxin and ferredoxin reductase efficiently hydroxylated 4-cholesten-3-one to the C-26 alcohol and subsequently to the acid. The X-ray structures of both substrate-free CYP125A3 and CYP142A2 and of cholest-4-en-3-one-bound CYP142A2 reveal significant differences in the substrate binding sites compared with the homologous M. tuberculosis proteins. Deletion only of cyp125A3 causes a reduction of both the alcohol and acid metabolites and a strong induction of cyp142 at the mRNA and protein levels, indicating that CYP142A2 serves as a functionally redundant back up enzyme for CYP125A3. In contrast to M. tuberculosis, the M. smegmatis Δcyp125Δcyp142 double mutant retains its ability to grow on cholesterol albeit with a diminished capacity, indicating an additional level of redundancy within its genome.

Published
2013

129. A new in vivo screening paradigm to accelerate antimalarial drug discovery.

Author

Jiménez-Díaz, María, Viera, Sara, Ibáñez, Javier, Mulet, Teresa, Magán-Marchal, Noemí, Garuti, Helen, Gómez, Vanessa, Cortés-Gil, Lorena, Martínez, Antonio, Ferrer, Santiago, Fraile, María, Calderón, Félix, Fernández, Esther, Shultz, Leonard, Leroy, Didier, García-Bustos, José, Gamo, Francisco, Angulo-Barturen, Iñigo, and Wilson, David

Subjects
Animals, Antimalarials, Drug Evaluation, Preclinical, Feasibility Studies, Female, Humans, Malaria, Mice, Parasitemia, Plasmodium berghei, Time Factors
Abstract

The emergence of resistance to available antimalarials requires the urgent development of new medicines. The recent disclosure of several thousand compounds active in vitro against the erythrocyte stage of Plasmodium falciparum has been a major breakthrough, though converting these hits into new medicines challenges current strategies. A new in vivo screening concept was evaluated as a strategy to increase the speed and efficiency of drug discovery projects in malaria. The new in vivo screening concept was developed based on human disease parameters, i.e. parasitemia in the peripheral blood of patients on hospital admission and parasite reduction ratio (PRR), which were allometrically down-scaled into P. berghei-infected mice. Mice with an initial parasitemia (P0) of 1.5% were treated orally for two consecutive days and parasitemia measured 24 h after the second dose. The assay was optimized for detection of compounds able to stop parasite replication (PRR = 1) or induce parasite clearance (PRR >1) with statistical power >99% using only two mice per experimental group. In the P. berghei in vivo screening assay, the PRR of a set of eleven antimalarials with different mechanisms of action correlated with human-equivalent data. Subsequently, 590 compounds from the Tres Cantos Antimalarial Set with activity in vitro against P. falciparum were tested at 50 mg/kg (orally) in an assay format that allowed the evaluation of hundreds of compounds per month. The rate of compounds with detectable efficacy was 11.2% and about one third of active compounds showed in vivo efficacy comparable with the most potent antimalarials used clinically. High-throughput, high-content in vivo screening could rapidly select new compounds, dramatically speeding up the discovery of new antimalarial medicines. A global multilateral collaborative project aimed at screening the significant chemical diversity within the antimalarial in vitro hits described in the literature is a feasible task.

Published
2013

134. AVALIAÇÃO DA ATIVIDADE ANTIMICOBACTERIANA IN VITRO DE ANÁLOGOS DA PODOFILOTOXINA CONTRA CEPAS DE MICOBACTÉRIAS NÃO TUBERCULOSAS

Author

Laet, João Paulo de Lucena, primary, Rodrigues, Danielle Martiniano da Silva, additional, Tejo, Renata Inglez de Souza, additional, da Silva, Kessia Kelly Batista, additional, de Lima, Milena Brandão, additional, Teixeira, Jéssica Lopes, additional, do Nascimento, Nathyeli Oliveira, additional, Rodrigues, Giovanna Gabriela Pedroza, additional, dos Santos Nascimento, Josefa Nayara, additional, Abrantes, Bárbara Wanessa Delgado, additional, Fernández, Esther Del Olmo, additional, Schindler, Haiana Charifker, additional, and Montenegro, Lilian Maria Lapa, additional

Published
2023
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135. AVALIAÇÃO IN VITRO DO POTENCIAL TERAPÊUTICO DE DERIVADOS DE PODOFILOTOXINA CONTRA CEPAS DE MYCOBACTERIUM TUBERCULOSIS: PERSPECTIVAS PARA O DESENVOLVIMENTO DE NOVOS AGENTES ANTIMICROBIANOS

Author

Rodrigues, Danielle Martiniano da Silva, primary, Laet, João Paulo de Lucena, additional, Tejo, Renata Inglez de Souza, additional, da Silva, Kessia Kelly Batista, additional, de Lima, Milena Brandão, additional, Teixeira, Jéssica Lopes, additional, Nascimento, Nathyeli Oliveira do, additional, Rodrigues, Giovanna Gabriela Pedroza, additional, dos Santos Nascimento, Josefa Nayara, additional, Abrantes, Bárbara Wanessa Delgado, additional, Fernández, Esther Del Olmo, additional, Montenegro, Lilian Maria Lapa, additional, and Schindler, Haiana Charifker, additional

Published
2023
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