Els Actors de l'Euroregió Pirineus-Mediterrània. Xarxes, percepcions i expectatives

Segons la nostra hipòtesi, la viabilitat de l'Euroregió Pirineus Mediterrània (EPM) dependrà sobretot de la seva capacitat de mobilitzar, d'implicar i de connectar els actors públics i privats dels diferents nivells territorials als quals s'adreça aquesta iniciativa. Per tant, el grau de coneixement i el posicionament dels diferents actors respecte al projecte resulten essencials a l'hora de definir una estratègia de cooperació d'aquestes característiques. L'objectiu d'aquest document és explicar els resultats de l'enquesta duta a terme amb un miler d'actors públics i privats de l'EPM. Aquesta tenia per objectiu obtenir dades relatives a aspectes clau del context en el que s'insereix el projecte, així com informació rellevant per definir estratègies d'actuació que poden ajudar a desenvolupar les capacitats organitzatives i comunicatives necessàries per gestionar una organització complexa com aquesta. L'estudi empíric es va estructurar al voltant de tres línies d'anàlisi: 1r) el present de l'Euroregió; 2n) El posicionament respecte l'EPM i 3r) El futur de l'EP

Rapid Differential Diagnosis between Extrapulmonary Tuberculosis and Focal Complications of Brucellosis Using a Multiplex Real-Time PCR Assay

BACKGROUND: Arduous to differ clinically, extrapulmonary tuberculosis and focal complications of brucellosis remain important causes of morbidity and mortality in many countries. We developed and applied a multiplex real-time PCR assay (M RT-PCR) for the simultaneous detection of Mycobacterium tuberculosis complex and Brucella spp. METHODOLOGY: Conventional microbiological techniques and M RT-PCR for M. tuberculosis complex and Brucella spp were performed on 45 clinical specimens from patients with focal complications of brucellosis or extrapulmonary tuberculosis and 26 control samples. Fragments of 207 bp and 164 bp from the conserved region of the genes coding for an immunogenic membrane protein of 31 kDa of B. abortus (BCSP31) and the intergenic region SenX3-RegX3 were used for the identification of Brucella and M. tuberculosis complex, respectively. CONCLUSIONS: The detection limit of the M RT-PCR was 2 genomes per reaction for both pathogens and the intra- and inter-assay coefficients of variation were 0.44% and 0.93% for Brucella and 0.58% and 1.12% for Mycobacterium. M RT-PCR correctly identified 42 of the 45 samples from patients with tuberculosis or brucellosis and was negative in all the controls. Thus, the overall sensitivity, specificity, PPV and NPV values of the M RT PCR assay were 93.3%, 100%, 100% and 89.7%, respectively, with an accuracy of 95.8% (95% CI, 91.1%-100%). Since M RT-PCR is highly reproducible and more rapid and sensitive than conventional microbiological tests, this technique could be a promising and practical approach for the differential diagnosis between extrapulmonary tuberculosis and focal complications of brucellosis

Real-time PCR optimization to identify Mycobacterium tuberculosis complex strains in clinical samples.

Resumen del artículo publicado en FEBS Journal,http://dx.doi.org/10.1111/febs.12919During recent years several molecular techniques have become available for Mycobacterium tuberculosis complex (MTC) detection,both for clinical samples and for isolates. One of the techniques more widely used is real time PCR in combination with nucleic acid amplification protocols. There are numerous studies based on PCR for the diagnosis of tuberculosis although the different protocols and primers used in the laboratory, together with the variability in the diagnostic performance of the methods tested, require that a comparative study be performed. Furthermore,the fact that the detection from clinical samples requires using highly sensitive targets suggests that this type of study should include multicopy targets to compare their efficiency with respect to the single copy. Our aim was to identify the members of the MTC using real-time PCR assays based on SYBR Green,among a large panel of isolated bacterial strains and clinical samples.We chose three targets (IS6110, senx3-regx3 and cfp32) and the optimal values for each PCR assay were empirically defined by testing in triplicate different concentrations of MgCl2 and primer sets and different annealing temperatures. These conditions were determined based on the specific amplification reactions that showed a lower Ct value, higher fluorescence and absence of non-specific PCR products. The analytical sensitivity was evaluated by ten-fold serial dilutions of DNA from MTC and the specificity was tested by 62 different microorganisms, including bacteria related with the MTC. The diagnostic yield was evaluated in 66 specimens from patients with suspected tuberculosis;30 had tuberculosis and 36 (control group) had different diseases.Under the conditions that resulted in optimization, standard curves showed that senx3-regx3 assay was the most efficient, followed by IS6110 and cfp32. However, the detection of bacterial DNA was faster with the repetitive element IS6110, with Ct values of up to 3 and 9 cycles of difference with respect to senx3-regx3 and cfp32. The analytical specificity, done only with the senx3-regx3 and IS6110 targets, was in the order of 100 and 93.5%, since IS6110 amplified various non-tuberculous micobacteria.For all the clinical samples studied, the sensitivity of both assays was identical (93.3%) but the specificity of senx3-regx3(100%) was higher than that of IS6110 (94.7%). In conclusion,real time PCR assay-SYBR Green based on the targets senx3-regx3 is highly reproducible and more sensitive and specific than the assays based on IS6110 or cfp32. The protocol developed in this study provides an appropriate and rapid tool to identify the strains of MTC in different clinical isolates and specimens.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Combining ability for yield among maize inbred lines obtained from novel sources of resistance to "Mal de Río Cuarto"

Los estudios de aptitud combinatoria proveen información sobre la capacidad de las líneas de maíz (Zea mays L.) para la formación de híbridos. Se realizó un experimento dialélico (Método 4 modelo I, Griffing, 1956) entre nueve líneas de maíz, derivadas de compuestos formados con nuevas fuentes de resistencia al mal de Río Cuarto, con el objeto de determinar la aptitud combinatoria general y específica (ACG y ACE) para rendimiento. Además, se evaluó vuelco y quebrado a fin de establecer el comportamiento de los híbridos para estas variables. Los ambientes de evaluación fueron las localidades Pergamino, Junín, Ferré (Buenos Aires), en la región maicera VI y las localidades Sampacho y Holmberg (Córdoba), en la región maicera IV. Considerando los efectos de ACG, la línea de mayor aporte al rendimiento fue 2600. Los híbridos que presentaron mejor ACE fueron 2600 × 2335 y 2568-2 × 2561. Los híbridos formados por LP561 presentaron bajos valores de vuelco y los formados por 2600 y LP561, bajos valores de quebrado. Los resultados de este trabajo indican que algunos de los híbridos formados por las nuevas líneas presentaron alto rendimiento y menores porcentajes de vuelco y quebrado en comparación con aquellos de híbridos formados por algunas de las líneas que habían sido usadas como fuentes de resistencia, y con aquellos mostrados por los testigos comerciales. Por lo tanto, estas líneas podrían ser usadas directamente en la formación de híbridos comerciales o en la creación de compuestos incrementando la variabilidad del germoplasma elite de maíz.Assessing combining ability of maize (Zea mays L.) inbred lines is important to predict hybrid performance. A diallel cross (Method 4 model I, Griffing, 1956) was performed among nine maize inbred lines, derived from composites formed with novel sources of resistance to Rio Cuarto disease, to assess general and specific combining ability (GCA & SCA) for yield. Others traits such as stalk and root lodging were measured. The selected environments were Pergamino, Junín and Ferré (Province of Buenos Aires), maize region # VI, and Sampacho and Holmberg (Province of Córdoba), maize region # IV. Inbred line 2600 showed the highest GCA for yield. Hybrids with the highest SCA for yield were 2600 × 2335 and 2568-2 × 2561. The hybrids formed with the inbred line LP561 exhibited low root lodging and the hybrids formed with 2600 and LP561 low stalk lodging. The results of these experiments indicate that some of the hybrids formed by the new inbred lines exhibited high yield and low percentages of root and stalk lodging compared to those of hybrids formed by some of the inbred lines that had been used as sources of resistance and to those of commercial hybrids. Therefore, these inbred lines might be directly used in hybrid formation or included in new breeding populations to increase the variability of elite maize germplasm.Facultad de Ciencias Agrarias y Forestale

Combining ability for yield among maize inbred lines obtained from novel sources of resistance to "Mal de Río Cuarto"

Los estudios de aptitud combinatoria proveen información sobre la capacidad de las líneas de maíz (Zea mays L.) para la formación de híbridos. Se realizó un experimento dialélico (Método 4 modelo I, Griffing, 1956) entre nueve líneas de maíz, derivadas de compuestos formados con nuevas fuentes de resistencia al mal de Río Cuarto, con el objeto de determinar la aptitud combinatoria general y específica (ACG y ACE) para rendimiento. Además, se evaluó vuelco y quebrado a fin de establecer el comportamiento de los híbridos para estas variables. Los ambientes de evaluación fueron las localidades Pergamino, Junín, Ferré (Buenos Aires), en la región maicera VI y las localidades Sampacho y Holmberg (Córdoba), en la región maicera IV. Considerando los efectos de ACG, la línea de mayor aporte al rendimiento fue 2600. Los híbridos que presentaron mejor ACE fueron 2600 × 2335 y 2568-2 × 2561. Los híbridos formados por LP561 presentaron bajos valores de vuelco y los formados por 2600 y LP561, bajos valores de quebrado. Los resultados de este trabajo indican que algunos de los híbridos formados por las nuevas líneas presentaron alto rendimiento y menores porcentajes de vuelco y quebrado en comparación con aquellos de híbridos formados por algunas de las líneas que habían sido usadas como fuentes de resistencia, y con aquellos mostrados por los testigos comerciales. Por lo tanto, estas líneas podrían ser usadas directamente en la formación de híbridos comerciales o en la creación de compuestos incrementando la variabilidad del germoplasma elite de maíz.Assessing combining ability of maize (Zea mays L.) inbred lines is important to predict hybrid performance. A diallel cross (Method 4 model I, Griffing, 1956) was performed among nine maize inbred lines, derived from composites formed with novel sources of resistance to Rio Cuarto disease, to assess general and specific combining ability (GCA & SCA) for yield. Others traits such as stalk and root lodging were measured. The selected environments were Pergamino, Junín and Ferré (Province of Buenos Aires), maize region # VI, and Sampacho and Holmberg (Province of Córdoba), maize region # IV. Inbred line 2600 showed the highest GCA for yield. Hybrids with the highest SCA for yield were 2600 × 2335 and 2568-2 × 2561. The hybrids formed with the inbred line LP561 exhibited low root lodging and the hybrids formed with 2600 and LP561 low stalk lodging. The results of these experiments indicate that some of the hybrids formed by the new inbred lines exhibited high yield and low percentages of root and stalk lodging compared to those of hybrids formed by some of the inbred lines that had been used as sources of resistance and to those of commercial hybrids. Therefore, these inbred lines might be directly used in hybrid formation or included in new breeding populations to increase the variability of elite maize germplasm.Facultad de Ciencias Agrarias y Forestale

Multifaceted role of cycling DOF factor 3 (CDF3) in the regulation of flowering time and abiotic stress responses in Arabidopsis

[EN] DNA-binding with one finger (DOF)-type transcription factors are involved in many fundamental processes in higher plants, from responses to light and phytohormones to flowering time and seed maturation, but their relation with abiotic stress tolerance is largely unknown. Here, we identify the roles of CDF3, an Arabidopsis DOF gene in abiotic stress responses and developmental processes like flowering time. CDF3 is highly induced by drought, extreme temperatures and abscisic acid treatment. The CDF3 T-DNA insertion mutant cdf3-1 is much more sensitive to drought and low temperature stress, whereas CDF3 overexpression enhances the tolerance of transgenic plants to drought, cold and osmotic stress and promotes late flowering. Transcriptome analysis revealed that CDF3 regulates a set of genes involved in cellular osmoprotection and oxidative stress, including the stress tolerance transcription factors CBFs, DREB2A and ZAT12, which involve both gigantea-dependent and independent pathways. Consistently, metabolite profiling disclosed that the total amount of some protective metabolites including -aminobutyric acid, proline, glutamine and sucrose were higher in CDF3-overexpressing plants. Taken together, these results indicate that CDF3 plays a multifaceted role acting on both flowering time and abiotic stress tolerance, in part by controlling the CBF/DREB2A-CRT/DRE and ZAT10/12 modules.We thank Dr Pablo Gonzalez-Melendi and Dr Jan Zouhar for technical handling of the confocal microscope and Dr Rafael Catala for the assistance with the low temperature stress assays. This work was supported by grants from Instituto Nacional de Investigacion y Tecnologia Agraria y Alimentaria (INIA; projects 2009-0004-C01, 2012-0008-C01), Spanish Ministry of Science and Innovation (projects BIO2010-1487, BFU2013-49665-EXP). A.R.C. and J.D.F. were supported by INIA pre-doctoral fellowshipsCorrales, AR.; Carrillo, L.; Lasierra, P.; Nebauer, SG.; Dominguez-Figueroa, J.; Renau-Morata, B.; Pollmann, S.... (2017). Multifaceted role of cycling DOF factor 3 (CDF3) in the regulation of flowering time and abiotic stress responses in Arabidopsis. Plant Cell & Environment. 40(5):748-764. https://doi.org/10.1111/pce.12894S748764405Achard, P., Gong, F., Cheminant, S., Alioua, M., Hedden, P., & Genschik, P. (2008). The Cold-Inducible CBF1 Factor–Dependent Signaling Pathway Modulates the Accumulation of the Growth-Repressing DELLA Proteins via Its Effect on Gibberellin Metabolism. The Plant Cell, 20(8), 2117-2129. doi:10.1105/tpc.108.058941Ahuja, I., de Vos, R. C. H., Bones, A. 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Cluster microclimate, canopy management and its influence on the berry (size and composition) quality

Sunlight and microclimate inside the clusters’ zone are key factors in berry development and must composition. Plant geometry and training system should be joined with a proper sunlight and temperature cluster microclimate and, also in the rest of the plant. Berry temperature can vary between 2 and 10 ºC or even more in inner clusters, depending on their exposure (Spayd et al., 2002). Sunlight, air ventilation within the canopy, temperature cluster and microclimate are affected by the exposure and radiation percentage received by grapes during its growth and maturation period (Deloire and Hunter 2005). In Mediterranean conditions (warm and dry climate), the use of porous systems may help plants establish a better leaf distribution inside this area (de la Fuente et al., 2015), providing more space and enhancing certain physiological processes, both in leaves (photosynthesis, ventilation, transpiration) and berries (growth and maturation). Grapes exposed to direct radiation are more sensitive to ripening and they can even suffer a dehydration process in the Mediterranean regions, where the temperature during the ripening after midday is frequently between 30-35 ºC or higher than 40 ºC (de la Fuente et al., 2015). A key point for well microclimate management inside the canopy is heat flux control, which is usually generated by three factors: surface area (SA) to PAR (direct or indirect) radiation; intensity or thermal value (related to the temperature) and time of exposure (de la Fuente, 2009; de la Fuente et al., 2013). Sprawl systems are non-positioned systems where vegetation is in multidirectional directions. Therefore, as sun position changes along the day, some leaves are first shaded and then others, so the sun leaf exposure decreases (Gutiérrez et al., 2021). The study was conducted in D.O. Uclés vineyards (lat. 39º50’8” N; long. 3º09’48.6” W; elevation 746 m above sea level) during the 2020 season, in cv. Tempranillo. The trial was designed with two training system: Vertical Shoot Positioned (VSP) and Sprawl (SP). Berry sampling was done every three days within the final 15 days before the estimated harvest date. A single sample comprised 100 berries collected from the clusters of the 10 selected vines in each block. Weight, size, must composition (reducing sugar, pH, acidity, volatile acidity, etc.) and skin composition (total and acylated monomeric pigments, TPI) were analysed. Regarding the berry composition, SP accumulated larger concentration of reducing sugars (+7.4%). No statistical differences were observed in the remaining oenological parameters measured in the berries between treatments. Nevertheless, inside total pigments and color parameters, TPI values reached significantly higher (+40%) in the SP vs VSP treatments. In the total concentration of pigments (including the acylated fraction) and pH, no differences were founded. Berry's weight and size showed some relevant differences between treatments. During the end maturity-harvest period, the berry weight (12-11%) and berry size (6-9%) were higher in SP treatment compared to VSP. These results suggest that the SP systems can induce an increment of reducing sugars, and TPI and also, can modulate the berry weight and size, helping to control overripening and berry dehydration processes. Therefore, sprawl systems (SP) represent an alternative to VSP systems in warm areas for achieving an increment of pigments, as well as for better control of the accumulation of reducing sugars, without compromising the harvest yield (higher berry weight and size)

La enseñanza del metabolismo: retos y oportunidades

En el marco del Proyecto de Innovación Educativa de la Universidad de Málaga PIE15-163, cuya descripción y resultados incluimos, decidimos que esta era una excelente oportunidad para reflexionar acerca de la enseñanza del metabolismo y de poner por escrito dichas reflexiones en un libro. Quisimos y pudimos contar con la colaboración de buena parte de los compañeros del Departamento de Biología Molecular y Bioquímica que apoyaron con su firma el proyecto PIE15-163 y extendimos nuestra invitaciones a otros compañeros de dentro y fuera de la Universidad de Málaga. Del Departamento de Biología Molecular y Bioquímica de la Universidad de Málaga hemos recibido aportaciones de los catedráticos Victoriano Valpuesta Fernández, Ana Rodríguez Quesada y Antonio Heredia Bayona, los profesores titulares María Josefa Pérez Rodríguez, José Luis Urdiales Ruiz e Ignacio Fajardo Paredes y la investigadora postdoctoral y profesora sustituta interina Beatriz Martínez Poveda. De otros departamentos de la Universidad de Málaga hemos contado con las aportaciones de la catedrática del Departamento de Especialidades Quirúrgicas, Bioquímica e Inmunología Pilar Morata Losa, del catedrático del Departamento de Lenguajes y Ciencias de la Computación José Francisco Aldana Montes y los componentes de su grupo de investigación Khaos Ismael Navas Delgado, María Jesús García Godoy, Esteban López Camacho y Maciej Rybinski, del catedrático Ángel Blanco López, del Área de Conocimiento de Didáctica de las Ciencias Experimentales y del Doctor en Ciencias Químicas y actual doctorando del Programa de Doctorado "Educación y Comunicación Social" Ángel Luis García Ponce. De fuera de la Universidad de Málaga, hemos contado con las aportaciones del catedrático de la Universidad de La Laguna Néstor V. Torres Darias, de la catedrática de la Universitat de les Illes Balears Pilar Roca Salom y de sus compañeros los profesores Jorge Sastre Serra y Jordi Oliver, de los catedráticos de la Universidad de Granada Rafael Salto González y María Dolores Girón González y su colaborador el Dr. José Dámaso Vílchez Rienda, del profesor titular de la Universidad de Alcalá Ángel Herráez, del investigador postdoctoral de la Universidad de Erlangen (Alemania) Guido Santos y del investigador postdoctoral de la empresa Brain Dynamics Carlos Rodríguez Caso.Hemos estructurado los contenidos del libro en diversas secciones. La primera presenta el Proyecto en cuyo marco se ha gestado la iniciativa que ha conducido a la edición del presente libro. La segunda sección la hemos titulado "¿Qué metabolismo?" e incluye diversas aportaciones personales que reflexionan acerca de qué metabolismo debe conocer un graduado en Bioquímica, en Biología, en Química, en Farmacia o en Medicina, así como una aportación acerca de qué bioquímica estructural y enzimología son útiles y necesarias para un estudiante que vaya a afrontar el estudio del metabolismo. La tercera sección, "Bases conceptuales", analiza las aportaciones del aprendizaje colaborativo, el contrato de aprendizaje y el aprendizaje basado en la resolución de casos prácticos a la mejora del proceso enseñanza-aprendizaje dentro del campo de la Bioquímica y Biología Molecular, más concretamente en el estudio del metabolismo. La cuarta sección se titula "Herramientas", es la más extensa e incluye las diversas aportaciones centradas en propuestas concretas de aplicación relevantes y útiles para la mejora de la docencia-aprendizaje del metabolismo. Sigue una sección dedicada a presentar de forma resumida los "Resultados" del proyecto PIE15-163. El libro concluye con una "coda final" en la que se reflexiona acerca del aprendizaje de la Química a la luz de la investigación didáctica.Patrocinado por el Proyecto de Innovación Educativa de la Universidad de Málaga PIE15-16