Búsqueda de dianas en Podosphaera Xanthii para el desarrollo de nuevas fitoterapias antifúngicas

La resistencia a fungicidas en uno de los principales problemas de la agricultura y esto es especialmente patente en el caso de los oídios. En España y en las principales áreas productoras del mundo, el oídio de las cucurbitáceas es una amenaza muy grave, y Podosphaera xanthii es considerado como el principal agente causante de la enfermedad. Hasta la fecha, la aplicación de fungicidas y el uso de variedades resistentes son las principales herramientas para el control de la enfermedad. En cualquier caso, el oídio sigue imponiendo serias limitaciones en la producción agrícola, siendo necesario el desarrollo de nuevas estrategias de control. En este estudio se pretende proporcionar información sobre las bases moleculares de P. xanthii que pueda ser de utilidad para el desarrollo de nuevas herramientas de fitoprotección. Para lograr este objetivo, estamos centrando nuestra atención en un conjunto de proteínas fúngicas carentes de función, determinado en un estudio anterior. Ante la falta de homología con proteínas funcionalmente anotadas, para conocer la posible función de dichas proteínas, en primer lugar, llevamos a cabo un análisis in silico detallado de las proteínas que incluye modelado 3D, predicción de posibles ligandos e identificación de dominios funcionales. En segundo lugar, para la identificación de proteínas clave para la patogénesis de P. xanthii, silenciamos proteínas del hongo con función predicha bioinformáticamente mediante silenciamiento génico inducido por hospedador (HIGS) empleando Agrobacterium tumefaciens como vector para la expresión transitoria de construcciones de silenciamiento en células de melón, y desde ellas, al hongo diana. Finalmente, las proteínas con un fenotipo claro de silenciamiento serán seleccionadas para la caracterización de su actividad biológica. En este congreso se mostrarán los resultados más relevantes obtenidos hasta la fecha, relativos a la asimilación de azufre.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Este trabajo ha sido financiado por una ayuda del Programa Estatal de I+D+i Orientada a los Retos de la Sociedad (AGL2016-76216-C2-1-R), cofinanciada con fondos FEDER (UE). Los autores agredecen además ayudas del Plan Propio de Investigación de la Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Proteínas secretadas por el haustorio y su importancia en el desarrollo de Podosphaera xanthii

El cultivo de las cucurbitáceas en España se ve afectado, entre otros problemas, por Podosphaera xanthii, un hongo biotrofo obligado y principal agente causal del oídio de las cucurbitáceas. Este hongo, que requiere células vivas de la planta para completar su ciclo de vida asexual, desarrolla unas estructuras especiales de parasitismo denominadas haustorios. Los haustorios prosperan dentro de las células epidérmicas y son responsables de la relación directa entre el patógeno y el hospedador, participando en la absorción de nutrientes de la planta y en la liberación de efectores en las células del huésped. Dada la estrecha relación entre los haustorios y las células vegetales, desarrollamos un transcriptoma haustorial de novo de P. xanthii y definimos su secretoma con la intención de encontrar proteínas secretadas que sean esenciales para la patogénesis. De esta manera pudimos identificar diferentes proteínas, la mayoría de ellas sin función anotada, expresadas específicamente en el haustorio. Gracias al modelado proteico obtuvimos información relevante sobre la posible función de alguna de estas proteínas secretadas sin anotación funcional y seleccionamos varias para su validación funcional mediante silenciamiento génico en el hospedador mediado por Agrobacterium (ATM-HIGS). Además, la expresión heteróloga de las proteínas seleccionadas y los subsecuentes ensayos enzimáticos nos ha permitido corroborar su actividad bioquímica predicha. Así, pudimos comprobar cómo proteínas secretadas específicamente por el haustorio son muy importantes para el desarrollo fúngico. Este abordaje multidisciplinar nos ha permitido identificar y dilucidar la función de proteínas esenciales para P. xanthii que, dado su papel clave, se convierten en dianas esperanzadoras para el desarrollo de nuevo fitosanitarios contra el oídio de las cucurbitáceas y otros hongos fitopatógenos.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Este trabajo ha sido financiado por el Ministerio de Economía y Competitividad (AGL2013-41939-R; AGL2016-76216-C2-1-R) cofinanciado con fondos FEDER (UE). Álvaro Polonio es beneficiario de un contrato predoctoral (BES-2014-068602) para la formación de doctores del Ministerio de Economía y Competitividad. Los autores también agradecen ayudas del Plan Propio de Investigación de la Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Evaluation of biological control agents for managing cucurbit powdery mildew on greenhouse-grown melon

An evaluation was made of the ability of two mycoparasite-based products AQ10® (Ampelomyces quisqualis) and Mycotal® (Lecanicillium lecanii), as well as three strains of Bacillus subtilis, to manage powdery mildew disease, caused by Podosphaera fusca on melon seedlings maintained under different regimes of relative humidity and on plants grown under greenhouse conditions in Spain. In every case fungal and bacterial biocontrol agents (BCAs) performed better under conditions of high relative humidity (90–95% RH). In greenhouse experiments, the effectiveness of the mycoparasites to manage powdery mildew was absolutely dependent on mineral oil. The strains of B. subtilis provided disease control similar to that achieved with the mycoparasites or the fungicide azoxystrobin. Microscopic analysis showed the ability of these bacterial strains to efficiently colonize leaf surfaces and revealed the occurrence of antagonistic interactions between biological agents and P. fusca structures. These results confirmed the usefulness of these BCAs for managing powdery mildew on greenhouse-grown cucurbits either as single products or as a component of integrated control programmes.Estación Experimental ‘La Mayora’ (CSIC), Algarrobo-Costa, 29750 Málaga, Spain Grupo de Microbiología y Patología Vegetal-Unidad Asociada a CSIC, Departamento de Microbiología, Facultad de Ciencias, Universidad de Málaga, 29071 MálagaPeer reviewe

Increasing Therapy Related Myeloid Neoplasms in Multiple Myeloma

© 2018 Stichting European Society for Clinical Investigation Journal Foundation. This document is made available under the CC-BY-NC 4.0 license http://creativecommons.org/licenses/by-nc /4.0/ This document is the submitted version of a published work that appeared in final form in European Journal of Clinical Investigation.Background: Despite the longer survival achieved in multiple myeloma (MM) patients due to new therapy strategies, a concern is emerging regarding an increased risk of secondary primary malignancies (SPMs) and how to characterize those patients at risk. We performed a retrospective study covering a 28‐year follow‐ up period (1991‐2018) in a tertiary single institution. Material and Methods: Data of 403 MM patients were recorded and compared with the epidemiologic register of the population area covered by our centre, calculating the standardize incidence ratio (SIR) for the different types of SPMs diagnosed in the MM cohort. Fine and Gray regression models were used to identify risk factors for SPMs. Results: Out of the 403 MM patients, 23 (5.7%) developed SPMs: 13 therapyrelated myeloid (TRM) malignancies (10 of them (77%) myelodysplastic syndrome (MDS), 1 acute lymphoid leukaemia and 9 solid neoplasms. In the MM cohort, the relative risk of MDS was significantly higher than in the general population. Survival of patients with TRM malignancies was poor with a median of 4 months from the diagnosis, and most of them showed complex karyotype. Within the MM subset, multivariable analysis showed a higher risk of TRM malignancies in patients that previously received prolonged treatment with lenalidomide (>18 months). Conclusions: Though the improvement in MM outcome during the last decades is an unprecedented achievement, it has been accompanied by the rise in TRM malignancies with complex cytogenetic profile and poor prognosis that are in the need of an improved biologic and therapeutic approach

Growth performance, in vitro antioxidant properties and chemical composition of the halophyte Limonium algarvense Erben are strongly influenced by the irrigation salinity

Limonium algarvense Erben (sea lavender) is a halophyte species with potential to provide natural ingredients with in vitro antioxidant, anti-inflammatory, neuroprotective and antidiabetic properties. This study reports for the first time the 1) cultivation of sea lavender in greenhouse conditions under irrigation with freshwater (approx. 0 mM NaCl) and saline aquaculture wastewater (300 and 600 mM NaCl), and 2) the influence of the irrigation salinity on the plant performance (e.g growth, number of produced leaves and flowers), in vitro antioxidant properties [radical scavenging activity (DPPH and ABTS), ferric reducing antioxidant power (FRAP), metal chelating properties on copper (CCA) and iron (ICA)], toxicity (in vitro on three mammalian cell lines) and chemical composition (determined by LC-ESI-HRMS/MS). The freshwater-irrigated plants had better growth performance than those irrigated with saltwater. Extracts from wild plants, had the highest antioxidant activity, but those from cultivated ones kept high in vitro antioxidant properties and interesting chemical profile. The flowers' extracts of plants irrigated with 300 mM NaCl had the highest antioxidant activities against DPPH, whereas those from freshwater-irrigated plants were more active on ABTS, CCA and FRAP. Most of the extracts showed nil toxicity. The flowers' extracts displayed the highest diversity of compounds, mainly quercetin, apigenin, luteolin, naringenin and their glycoside derivatives. Moreover, their abundance varied with the irrigation salinity. These data indicate that sea lavender plants can be successfully cultivated in greenhouse conditions under fresh- and saltwater irrigation, maintaining interesting biological and chemical properties.Funding Agency Portuguese Foundation for Science and Technology Portuguese National Budget CCMAR/Multi/04326/2019 GreenVet project ALG-01-0145-FEDER-028876 XtrerneAquaCrops FA-05-2017-028 Lisboa-01-0145-FEDER-022125-RNEM-IST ID/QUI/00100/201 Portuguese Foundation for Science and Technology SFRH/BD/116604/2016 CEECIND/00425/2017info:eu-repo/semantics/publishedVersio

A crowdsourcing database for the copy-number variation of the spanish population

Background: Despite being a very common type of genetic variation, the distribution of copy-number variations (CNVs) in the population is still poorly understood. The knowledge of the genetic variability, especially at the level of the local population, is a critical factor for distinguishing pathogenic from non-pathogenic variation in the discovery of new disease variants. Results: Here, we present the SPAnish Copy Number Alterations Collaborative Server (SPACNACS), which currently contains copy number variation profiles obtained from more than 400 genomes and exomes of unrelated Spanish individuals. By means of a collaborative crowdsourcing effort whole genome and whole exome sequencing data, produced by local genomic projects and for other purposes, is continuously collected. Once checked both, the Spanish ancestry and the lack of kinship with other individuals in the SPACNACS, the CNVs are inferred for these sequences and they are used to populate the database. A web interface allows querying the database with different filters that include ICD10 upper categories. This allows discarding samples from the disease under study and obtaining pseudo-control CNV profiles from the local population. We also show here additional studies on the local impact of CNVs in some phenotypes and on pharmacogenomic variants. SPACNACS can be accessed at: http://csvs.clinbioinfosspa.es/spacnacs/. Conclusion: SPACNACS facilitates disease gene discovery by providing detailed information of the local variability of the population and exemplifies how to reuse genomic data produced for other purposes to build a local reference database.This work is supported by Grants PID2020-117979RB-I00 from the Spanish Ministry of Science and Innovation; by the Institute of Health Carlos III (project IMPaCT-Data, exp. IMP/00019, IMP/00009 and PI20/01305), co-funded by the European Union, European Regional Development Fund (ERDF, “A way to make Europe”)

Does co-infection with vector-borne pathogens play a role in clinical canine leishmaniosis?

The severity of canine leishmaniosis (CanL) due to Leishmania infantum might be affected by other vector-borne organisms that mimic its clinical signs and clinicopathological abnormalities. The aim of this study was to determine co-infections with other vector-borne pathogens based on serological and molecular techniques in dogs with clinical leishmaniosis living in Spain and to associate them with clinical signs and clinicopathological abnormalities as well as disease severity. Sixty-one dogs with clinical leishmaniosis and 16 apparently healthy dogs were tested for Rickettsia conorii, Ehrlichia canis, Anaplasma phagocytophilum and Bartonella henselae antigens by the immunofluorescence antibody test (IFAT) and for E. canis, Anaplasma spp., Hepatozoon spp., Babesia spp. and filarioid DNA by polymerase chain reaction (PCR). Among the dogs examined by IFAT, the seroprevalences were: 69% for R. conorii, 57% for E. canis, 44% for A. phagocytophilum and 37% for B. henselae ; while the prevalences found by PCR were: 8% for Ehrlichia / Anaplasma, 3% for Anaplasma platys and 1% for H. canis. No other pathogen DNA was detected. Statistical association was found between dogs with clinical leishmaniosis and seroreactivity to R. conorii antigen (Fisher's exact test: P = 0.025, OR = 4.1, 95% CI = 1-17) and A. phagocytophilum antigen (Fisher's exact test: P = 0.002, OR = 14.3, 95% CI = 2-626) and being positive to more than one serological or molecular tests (co-infections) (Mann-Whitney test: U = 243, Z = -2.6, n = 14, n = 61, P = 0.01) when compared with healthy dogs. Interestingly, a statistical association was found between the presence of R. conorii, E. canis, A. phagocytophilum and B. henselae antibodies in sick dogs and some clinicopathological abnormalities such as albumin and albumin/globulin ratio decrease and increase in serum globulins. Furthermore, seroreactivity with A. phagocytophilum antigens was statistically associated with CanL clinical stages III and IV. This study demonstrates that dogs with clinical leishmaniosis from Catalonia (Spain) have a higher rate of co-infections with other vector-borne pathogens when compared with healthy controls. Furthermore, positivity to some vector-borne pathogens was associated with more marked clinicopathological abnormalities as well as disease severity with CanL