Geomorphological setting and main technological features of new Middle and Upper Pleistocene sites in the Lower Manzanares River Valley (Madrid, Spain)

Las intervenciones arqueológicas llevadas a cabo durante los años 1996 en Tafesa, 2005 en el yacimiento Hospital 12 de Octubre y 2006 en la desembocadura del arroyo Butarque (Villaverde-Barrio de Butarque) situados al sur de la ciudad de Madrid (España), han aportado nuevos conjuntos líticos contextualizados estratigráficamente en los depósitos fluviales pleistocenos correspondientes al tramo inferior del valle del río Manzanares. Los yacimientos arqueológicos analizados se sitúan geomorfológicamente en la denominada “Terraza Compleja del Manzanares” (TCMZ), la cual constituye un nivel fluvial engrosado (20-15 m de potencia) situado entre +22-16 m sobre el cauce actual del río, a lo largo de su margen derecha. Este nivel fluvial ha sido tradicionalmente considerado de edad Pleistoceno medio en base a la industria achelense y complejos faunísticos encontrados en sus niveles inferiores. Ciertamente, Tafesa es un yacimiento situado en la parte inferior-media de la terraza de +22 m con industria achelense y fauna de Pleistoceno medio. Por el contrario, los niveles superiores de esta misma terraza en los sectores del 12 de Octubre y Villaverde-Butarque se encuentran asociados a industrias del Paleolítico inferior y medio ya pertenecientes al Pleistoceno superior, como sugieren el conjunto de dataciones OSL y TL existentes para la zona. Los datos analizados en este trabajo indican que el desarrollo de este nivel de terraza engrosado comienza durante la parte final de Pleistoceno medio y abarca todo el Estadio Isotópico OIS 5, ya dentro del Pleistoceno superiorThe archaeological works developed during the years 1996 in the site of Tafesa, 2005 in the 12 de Octubre Metro Station site and 2006 in the confluence of the Butarque Stream (Villaverde-Barrio de Butarque site) located south of the Madrid City (Spain), have provided new lithic assemblages. These assemblages have been stratigraphically contextualized in the Pleistocene deposits of the Lower Manzanares river valley within the so-called “Manzanares Complex Terrace” (TCMZ). This fluvial terrace constitutes an anomalous thickened (20-15m) deposit at +22-16m above the present river thalweg mainly developed along the right (southern) valley margin. This fluvial level has been traditionally considered of middle Pleistocene age on the basis of the acheulian lithics and faunal assemblages typically located within its lower stratigraphic layers. Certainly, the Tafesa is a fluvial terrace site at +22 m with acheulian industry and middle Pleistocene faunal remains at its lower sedimentary sequence. However, the upper sedimentary levels of this same terrace in the 12 de Octubre y Villaverde-Butarque sites throw lithic assemblages of the lower and upper Paleolithic belonging to upper Pleistocene, as suggested by the available set of TL and OSL dates for the zone. The analyses developed in this study indicate that the development of this thickened fluvial terrace started during the end of the middle Pleistocene, but also comprise the whole Oxygen Isotopic Stage OIS 5 during the upper Pleistocene

Inactivation of the Sinorhizobium fredii HH103 rhcJ gene abolishes nodulation outer proteins (Nops) secretion and decreases the symbiotic capacity with soybean

It has been postulated that nodulation outer proteins (Nops) avoid effective nodulation of Sinorhizobium fredii USDA257 to nodulate with American soybeans. S. fredii HH103 naturally nodulates with both Asiatic (non-commercial) and American (commercial) soybeans. Inactivation of the S. fredii HH103 gene rhcJ, which belongs to the tts (type III secretion) cluster, abolished Nop secretion and decreased its symbiotic capacity with the two varieties of soybeans. S. fredii strains HH103 and USDA257, that only nodulates with Asian soybeans, showed different SDS-PAGE Nop profiles, indicating that these strains secrete different sets of Nops. In coinoculation experiments, the presence of strain USDA257 provoked a clear reduction of the nodulation ability of strain HH103 with the American soybean cultivar Williams. These results suggest that S. fredii Nops can act as either detrimental or beneficial symbiotic factors in a strain-cultivar-dependent manner. Differences in the flavonoid-mediated expression of rhcJ with respect to nodA were also detected. In addition, one of the Nops secreted by strain HH103 was identified as NopA. [Int Microbiol 2006; 9(2):125-133

Quaternary fossil horses within the Prados-Guatén Depression (Pantoja de La Sagra, Toledo)

Durante la primera reunión de campo del Grupo Madrileño de Cuaternario (GQM-AEQUA) se localizaron restos fragmentarios de dentición de caballos fósiles en los antiguos areneros de Pantoja de La Sagra (Toledo), actualmente en proceso de desmantelamiento y relleno. Ante la posibilidad de deterioro y pérdida los restos fueron recolectados y trasladados al Museo Nacional de Ciencias Naturales (CSIC, Madrid) donde se ha procedido a su análisis. Las piezas fósiles analizadas responden a un maxilar izquierdo con tres piezas dentales in situ (molares y premolares), y otras siete más aisladas. Todos los dientes aislados, junto con el fragmento de maxilar existente, corresponden a un adulto joven. Los restos fósiles se encontraban asociados a un nivel de arenas fluviales situado unos cuatro metros por debajo de la superficie de la Terraza de +15 m de la Depresión Prados-Guatén definida como un nivel perteneciente al tránsito Pleistoceno inferior-medio, del antiguo Sistema fluvial Manzanares-Guatén por Silva (1988). En concreto los niveles superiores de esta terraza han sido interpretados como resultado de la superposición de los últimos depósitos del antiguo sistema fluvial y los primeros asociados al relleno de la Depresión por tributarios de área fuente más local tras su abandono como consecuencia del proceso de captura del valle inferior del Manzanares por parte del Río Jarama al SW de la Ciudad de Madrid (Silva et al., 1988). Los caracteres morfológicos y morfométricos de las piezas dentarias permiten identificarlos como Equus ferus cf. mosbachensis cuya distribución bioestratigráfica abarca la parte final del Pleistoceno Medio (c.a. 500-200 ka B.P.). Junto a los restos fósiles aparecieron también escasos fragmentos líticos correspondientes a productos de lascado en sílex de difícil atribución tecnológica. Los restos fósiles analizados, indican que el depósito extensivo de arenas fluviales en el eje de la Depresión, culminó durante el final del Pleistoceno medio, y que la dinámica fluvial de la Depresión tras su proceso de abandono fue de hecho más activa de lo que se pensaba con la instalación de sistemas de arroyos relevantes alimentados por cabeceras locales antes del encajamiento definitivo actual de los arroyos Prados y Guatén.During the first field-meeting of the Madrid Quaternary Research Group (GQM-AEQUA) several fossil teeth remnants of horses were localised at the ancient sand-quarries of Pantoja de La Sagra (Toledo), which presently are abandoned and refilling in progress. The possibility of deterioration and loss of the localised fossils remnants induced by the quarry works, they were collected and taken away to the Museo Nacional de Ciencias Naturales (CSIC, Madrid) for their preservation and analysis. Fossil remains correspond to a left maxilla with two in situ molars, another one inset on its alveolar cavity, fragments of premolar cavities, as well as other seven more isolated teeth. These fossils were outcropping in a sandy level at four meters below the +15 m fluvial terrace surface of the axial sector of de Prados-Guatén Depression, which is considered the last fluvial level belonging to the ancient Manzanares-Guatén fluvial system during the Lower-Middle Pleistocene transit (Silva, 1988). In detail, the upper fluvial sediments of this particular terrace level were interpreted as the result of the overlapping between the last materials deposited by the ancient Manzanares-Guatén fluvial system and the first ones resulting from the readjustment of former tributaries after the abandonment of the Depression caused by fluvial capture of the Lower Manzanares Valley SW Madrid City. The morphological features of the oclusal surface of the horse teeth and morphometric comparative analyses indicate that they belong to the specie Equus ferus, and probably to the subspecie mosbachensis. However due to the bad definition of this group in Europe and the few individuals analysed the better classification is Equus ferus cf. mosbachensis. The bioestratigraphic distribution of this fossil horse group in Europe extends on the upper part of the Middle Pleistocene (c.a. 500-200 ka B.P.). Few lithic artefacts outcropped also associated to the fossil remains, constituted by laminar flakes of hard technological classification. Fossil remains analysed in this work joint to the unique previous quaternary fossil mammal described for the Prados-Guatén Depression constituted by Mammuthus meridionalis NESTI of the former quarry of Esquivias adjacent to the AVE railway line (Silva et al., 1988b; 1999). The chronostratigraphic attribution of the fossil horses (Upper Middle Pleistocene) described here indicate that fluvial sedimentary activity within the Depression was relevant after its abandonment. Ancient tributaries of the former Manzanares-Guatén fluvial system, feed by local-intrabasinal headwaters, reworked the previous sandy sediments triggering multiepisodic deposition during the upper part of the Middle Pleistocene, before the more recent eventual incision of present streams dissecting the Depression

Dynamics and Mechanical Stability of the Developing Dorsoventral Organizer of the Wing Imaginal Disc

Shaping the primordia during development relies on forces and mechanisms able to control cell segregation. In the imaginal discs of Drosophila the cellular populations that will give rise to the dorsal and ventral parts on the wing blade are segregated and do not intermingle. A cellular population that becomes specified by the boundary of the dorsal and ventral cellular domains, the so-called organizer, controls this process. In this paper we study the dynamics and stability of the dorsal-ventral organizer of the wing imaginal disc of Drosophila as cell proliferation advances. Our approach is based on a vertex model to perform in silico experiments that are fully dynamical and take into account the available experimental data such as: cell packing properties, orientation of the cellular divisions, response upon membrane ablation, and robustness to mechanical perturbations induced by fast growing clones. Our results shed light on the complex interplay between the cytoskeleton mechanics, the cell cycle, the cell growth, and the cellular interactions in order to shape the dorsal-ventral organizer as a robust source of positional information and a lineage controller. Specifically, we elucidate the necessary and sufficient ingredients that enforce its functionality: distinctive mechanical properties, including increased tension, longer cell cycle duration, and a cleavage criterion that satisfies the Hertwig rule. Our results provide novel insights into the developmental mechanisms that drive the dynamics of the DV organizer and set a definition of the so-called Notch fence model in quantitative terms

A school-based physical activity promotion intervention in children: rationale and study protocol for the PREVIENE Project

The lack of physical activity and increasing time spent in sedentary behaviours during childhood place importance on developing low cost, easy-toimplement school-based interventions to increase physical activity among children. The PREVIENE Project will evaluate the effectiveness of five innovative, simple, and feasible interventions (active commuting to/from school, active Physical Education lessons, active school recess, sleep health promotion, and an integrated program incorporating all 4 interventions) to improve physical activity, fitness, anthropometry, sleep health, academic achievement, and health-related quality of life in primary school children. The PREVIENE Project will provide the information about the effectiveness and implementation of different school-based interventions for physical activity promotion in primary school children.The PREVIENE Project was funded by the Spanish Ministry of Economy and Competitiveness (DEP2015-63988-R, MINECO-FEDER). MAG is supported by grants from the Spanish Ministry of Economy and Competitivenes

The Expression of Heat Shock Protein HSP60A Reveals a Dynamic Mitochondrial Pattern in Drosophila melanogaster Embryos

The evolutionarily conserved hsp60 (heat-shock protein 60) family of molecular chaperones ensures the correct folding of nuclear-encoded proteins after their translocation across the mitochondrial membrane during development as well as after heat-shock treatment. Although the overexpression of HSP60 proteins and their localization in the cytoplasm have been linked with many humans pathologies, the detailed pattern of their expression in different animal models and their subcellular localization during normal development and in stress conditions are little-known. In this report, we have used two-dimensional gel electrophoresis followed by MALDI-TOF to identify and purify heat shock protein HSP60A of Drosophila melanoagaster. We demonstrate that it is heat-shock inducible and describe two novel antisera, specifically designed to recognize the denatured and native polypeptide, respectively, in Drosophila. Immunoelectron microscopy and immunostaining of Drosophila cells with these antibodies reveals that HSP60A is always localized to the inner membrane of mitochondria. Expression of HSP60A is post-transcriptionally regulated in a highly dynamic pattern during embryogenesis, even under heat-shock conditions. In contrast, in very stressful situations, its expression is upregulated transcriptionally over the entire embryo. These findings suggest novel roles for HSP60 family proteins during normal Drosophila development.This work was supported by Grant PB87-0449 from the Comisión Asesora de Investigación Científica y Técnica and by an institutional grant from Fundación Ramón Areces. J.A. was the recipient of a predoctoral fellowship of the Universidad Autónoma de Madrid. L.A.B.-L. was supported at various stages of the work by grants from the Dirección General de Investigación Científica y Técnica Grants and also by a postdoctoral fellowship from EMBO.Peer reviewe

Characterization of the Drosophila melanogaster mitochondrial proteome

We have combined high-resolution two-dimensional (2-D) gel electrophoresis with mass spectrometry with the aim of identifying proteins represented in the 2-D gel database of Drosophila melanogaster mitochondria. First, we purified mitochondria from third instar Drosophila larvae and constructed a high-resolution 2-D gel database containing 231 silver-stained polypeptides. Next, we carried out preparative 2-D PAGE to isolate some of the polypeptides and characterize them by MALDI-TOF analysis. Using this strategy, we identified 66 mitochondrial spots in the database, and in each case confirmed their identity by MALDI-TOF/TOF analysis. In addition, we generated antibodies against two of the mitochondrial proteins as tools for characterizing the organelle. © 2005 American Chemical Society.Comision Asesora de Investigación Científica y Técnica and by an institutional grant from Fundación Ramón ArecesPeer Reviewe

Constitutive expression of heat shock protein p23 correlates with proneural territories in imaginal discs of Drosophila melanogaster

2-DE followed by MALDI-TOF was used to purify and identify a Drosophila protein (catalogued as SSP 6002) that showed marked differences in the level of expression in the different imaginal discs of third instar larvae. Fingerprinting showed that the spot of interest was the heat shock 23 polypeptide (hsp23). We characterized the kinetics of its induction by heat shock in wing imaginal discs and raised an antiserum against the denatured protein, which recognizes a single unphosphorylated spot on 2-D gels. The difference in its expression in discs was corroborated by analyzing its level in the imaginal discs of postbithorax mutants. We also investigated the developmental expression of hsp23 in imaginal discs with antiserum raised against the native protein. Its spatial and temporal pattern of expression is related to the proneural territories and maintained even under heat shock conditions. In addition, its pattern of expression is regulated by transcription factors and signaling pathways (notch and epidermal growth factor receptor) involved in proneural specification. © 2005 Wiley-VCH Verlag GmbH & Co. KGaA.Comision Asesora de Investigación Científica y Técnica and by an institutional grant from Fundación Ramón ArecesPeer Reviewe

Unraveling plant cellular targets for the Rhizobium-specific effectors NopL and NopP

Resumen del póster presentado al 12th European Nitrogen Fixation Conference, celebrado en Budapest (Hungria) del 25 al 28 de agosto de 2016.Pathogenic Gram-negative use a specialized apparatus called Type 3 secretion system (T3SS) to deliver effectors directly into the eukaryotic host cells. These effectors suppress plant defenses to promote disease but they can also be recognized by specific plant receptors that trigger a strong defense reaction to eliminate the pathogen. The T3SS has also been found in some symbiotic rhizobial strains and the effectors secreted are involved in host-range determination and symbiotic efficiency. The broad host-range bacterium S. fredii HH103 secretes two proteins through the T3SS, NopL and NopP, which are specific for rhizobia. In this work we studied the function of both effectors in the symbiosis with soybean, which is considered its natural host plant. NopL and NopP were phosphorylated by soybean root kinases and the phosphorylation cascade was Ca2+ - and calmodulin-dependent. While the signaling pathway that culminates in the phosphorylation of NopL included ser/thr and MAPKK kinases, in the case of NopP this pathway involved ser/thr and tyr kinases but not MAPKK kinases. Transient expression of both nopL and nopP fused to YFP in Nicotiana benthamiana leaves and further confocal imaging indicated that they localized to the nucleus of the host cell. The use of a yeast-based array to determine possible effectors functions indicated that NopP could be involved in nuclear localization and migration. Finally, co-immunoprecipitation analyses of N. benthamiana NopL- and NopP-interacting proteins and further mass spectrometry analyses identified several potential plant targets for these effectors. The most interesting interactions are currently being validated by Bimolecfular Fluorescence Complementation (BiFC).This work was supported by project P11-CVI-7050 of the Junta de Andalucía.Peer Reviewe