Efecto De La Fibra De Polipropileno En El Comportamiento De La Mezcla Asfáltica En Caliente, Trujillo 2018

Esta investigación consistió en un análisis experimental de una mezcla asfáltica en caliente modificada con fibra de polipropileno, buscando mejorar su comportamiento ante los efectos del tránsito. Se evaluó el efecto de la fibra de polipropileno en el comportamiento de la mezcla asfáltica en caliente y para ello se realizaron ensayos de cántabro (pérdida por desgaste), estabilidad y flujo. Los materiales utilizados para la mezcla fueron, cemento asfáltico PEN 60/70 de la planta “Asfaltos Company Vial S.A.C” y agregados obtenidos de la cantera San Martin, provincia de Ascope, La Libertad a los cuales se les realizó una serie de ensayos de laboratorio. Los porcentajes de granulometría para la mezcla asfáltica fue 48% de agregado grueso, 50% de agregado fino, 2% de cal hidráulica y cemento asfáltico PEN 60/70 (4.5, 5.0, 5.5, 6.0 %), el diseño de la mezcla se realizó a través de la metodología Marshall en la ciudad de Trujillo. Los resultados que se obtuvieron de la evaluación de los especímenes de acuerdo al MTC E 504, fue un 5.3 % de cemento asfáltico óptimo y una estabilidad de 1215 kg con un flujo de 3.35 mm, por otro lado, en mezcla modificada con 0.5, 1.5, 2.5 % de fibra de polipropileno, se obtuvieron estabilidades de 1154, 1271, 1141 kg y flujo de 2.95, 3.19, 3.46 mm por cada porcentaje de fibra, siendo la estabilidad adecuada obtenida de la curva de 1270 kg y un flujo de 3.16 mm con un porcentaje óptimo de fibra de 1.4 %. Los resultados obtenidos del ensayo MTC E 515, la pérdida por desgaste para los grupos experimentales con 0.5, 1.5, 2.5 % de fibra de polipropileno resulto 3.17, 3.04, 3.36 % respectivamente y 4.0 % para el grupo control. En esta investigación se concluyó que la mezcla modificada con 1.5% de fibra de polipropileno presenta menor pérdida por desgaste

Propiedades psicométricas del Coronavirus Anxiety Scale (CAS) en adolescentes, Chiclayo 2022

El presente estudio estableció como objetivo general: determinar las propiedades psicométricas del Coronavirus Anxiety Scale (CAS) en adolescentes de la ciudad de Chiclayo. La investigación fue aplicada de tipo instrumental y con diseño psicométrico; la muestra fue seleccionada a través de un muestreo no probabilístico por conveniencia, la misma que estuvo conformada por 384 adolescentes entre hombres y mujeres pertenecientes a una institución educativa de Chiclayo. Los principales resultados demostraron que se logró evidenciar adecuadas propiedades psicométricas de la Coronavirus Anxiety Scale (CAS). Con respecto a la validez de contenido se optó por el método juicio de expertos y el coeficiente V de Aiken el cual arrojó valores aceptables (>0.90). Además, se evidenció que los 5 ítems mostraron adecuados índices de asimetría y curtosis (+/-1.5); además adecuados índices de homogeneidad (>30); y cargas factoriales (>0.30). En lo que concierne a la validez de constructo, se demostró que el instrumento unidimensional compuesto por 5 reactivos presenta adecuados índices de bondad de ajuste. Por último, a través del coeficiente Alpha y Omega, se obtuvo un valor de 0,86, lo que demuestra una excelente confiabilidad del Coronavirus Anxiety Scale (CAS)

The role of a class III gibberellin 2-oxidase in tomato internode elongation

[EN] A network of environmental inputs and internal signaling controls plant growth, development and organ elongation. In particular, the growth-promoting hormone gibberellin (GA) has been shown to play a significant role in organ elongation. The use of tomato as a model organism to study elongation presents an opportunity to study the genetic control of internode-specific elongation in a eudicot species with a sympodial growth habit and substantial internodes that can and do respond to external stimuli. To investigate internode elongation, a mutant with an elongated hypocotyl and internodes but wild-type petioles was identified through a forward genetic screen. In addition to stem-specific elongation, this mutant, named tomato internode elongated -1 (tie-1) is more sensitive to the GA biosynthetic inhibitor paclobutrazol and has altered levels of intermediate and bioactive GAs compared with wild-type plants. The mutation responsible for the internode elongation phenotype was mapped to GA2oxidase 7, a class III GA 2-oxidase in the GA biosynthetic pathway, through a bulked segregant analysis and bioinformatic pipeline, and confirmed by transgenic complementation. Furthermore, bacterially expressed recombinant TIE protein was shown to have bona fide GA 2-oxidase activity. These results define a critical role for this gene in internode elongation and are significant because they further the understanding of the role of GA biosynthetic genes in organ-specific elongation.This work used the Vincent J. Coates Genomics Sequencing Laboratory at UC Berkeley, supported by NIH S10 Instrumentation Grants S10RR029668 and S10RR027303. We thank the Tomato Genetics Resource Center for providing seed of the M82 and Heinz cultivars. The material was developed by and/or obtained from the UC Davis/C M Rick Tomato Genetics Resource Center and maintained by the Department of Plant Sciences, University of California, Davis, CA 95616, USA. We thank Anthony Bolger, Alisdair Fernie and Bjorn Usadel for providing us with access to pre-publication genomic reads of the S. lycopersicum cultivar M82, and Cristina Urbez and Noel Blanco-Tourinan (IBMCP, Spain) for technical help with in vitro production of TIE1. This work was supported in part by the Elsie Taylor Stocking Memorial Fellowship awarded to ASL in 2013, by NSF grant IOS-0820854, by USDA National Institute of Food and Agriculture project CA-D-PLB-2465-H, by internal UC Davis funds, and by Spanish Ministry of Economy and Competitiveness grant BFU2016-80621-P.Lavelle, A.; Gath, N.; Devisetty, U.; Carrera Bergua, E.; Lopez Diaz, I.; Blazquez Rodriguez, MA.; Maloof, J. (2018). The role of a class III gibberellin 2-oxidase in tomato internode elongation. The Plant Journal. https://doi.org/10.1111/tpj.14145SAndrés, F., Porri, A., Torti, S., Mateos, J., Romera-Branchat, M., García-Martínez, J. L., … Coupland, G. (2014). SHORT VEGETATIVE PHASE reduces gibberellin biosynthesis at theArabidopsisshoot apex to regulate the floral transition. Proceedings of the National Academy of Sciences, 111(26), E2760-E2769. doi:10.1073/pnas.1409567111Bolger, A., Scossa, F., Bolger, M. E., Lanz, C., Maumus, F., Tohge, T., … Fernie, A. R. (2014). The genome of the stress-tolerant wild tomato species Solanum pennellii. Nature Genetics, 46(9), 1034-1038. doi:10.1038/ng.3046Bowen, M. E., Henke, K., Siegfried, K. R., Warman, M. L., & Harris, M. P. (2011). Efficient Mapping and Cloning of Mutations in Zebrafish by Low-Coverage Whole-Genome Sequencing. Genetics, 190(3), 1017-1024. doi:10.1534/genetics.111.136069Burset, M. (2000). Analysis of canonical and non-canonical splice sites in mammalian genomes. Nucleic Acids Research, 28(21), 4364-4375. doi:10.1093/nar/28.21.4364Chen, W., Yao, J., Chu, L., Yuan, Z., Li, Y., & Zhang, Y. (2015). Genetic mapping of the nulliplex-branch gene (gb_nb1) in cotton using next-generation sequencing. Theoretical and Applied Genetics, 128(3), 539-547. doi:10.1007/s00122-014-2452-2Cingolani, P., Platts, A., Wang, L. L., Coon, M., Nguyen, T., Wang, L., … Ruden, D. M. (2012). A program for annotating and predicting the effects of single nucleotide polymorphisms, SnpEff. Fly, 6(2), 80-92. doi:10.4161/fly.19695Cuperus, J. T., Montgomery, T. A., Fahlgren, N., Burke, R. T., Townsend, T., Sullivan, C. M., & Carrington, J. C. (2009). Identification of MIR390a precursor processing-defective mutants in Arabidopsis by direct genome sequencing. Proceedings of the National Academy of Sciences, 107(1), 466-471. doi:10.1073/pnas.0913203107Curtis, M. D., & Grossniklaus, U. (2003). A Gateway Cloning Vector Set for High-Throughput Functional Analysis of Genes in Planta. Plant Physiology, 133(2), 462-469. doi:10.1104/pp.103.027979Devisetty, U. K., Covington, M. F., Tat, A. V., Lekkala, S., & Maloof, J. N. (2014). Polymorphism Identification and Improved Genome Annotation ofBrassica rapaThrough Deep RNA Sequencing. G3: Genes|Genomes|Genetics, 4(11), 2065-2078. doi:10.1534/g3.114.012526Eckardt, N. A. (2007). GA Perception and Signal Transduction: Molecular Interactions of the GA Receptor GID1 with GA and the DELLA Protein SLR1 in Rice. The Plant Cell, 19(7), 2095-2097. doi:10.1105/tpc.107.054916Ernst, H. A., Lo Leggio, L., Willemoës, M., Leonard, G., Blum, P., & Larsen, S. (2006). Structure of the Sulfolobus solfataricus α-Glucosidase: Implications for Domain Conservation and Substrate Recognition in GH31. Journal of Molecular Biology, 358(4), 1106-1124. doi:10.1016/j.jmb.2006.02.056Fillatti, J. J., Kiser, J., Rose, R., & Comai, L. (1987). Efficient Transfer of a Glyphosate Tolerance Gene into Tomato Using a Binary Agrobacterium Tumefaciens Vector. Nature Biotechnology, 5(7), 726-730. doi:10.1038/nbt0787-726Garrison , E. Marth , G. 2012 Haplotype-based variant detection from short-read sequencingHedden, P., & Graebe, J. E. (1985). Inhibition of gibberellin biosynthesis by paclobutrazol in cell-free homogenates ofCucurbita maxima endosperm andMalus pumila embryos. Journal of Plant Growth Regulation, 4(1-4), 111-122. doi:10.1007/bf02266949Kimura, S., & Sinha, N. (2008). Tomato (Solanum lycopersicum): A Model Fruit-Bearing Crop. Cold Spring Harbor Protocols, 2008(12), pdb.emo105-pdb.emo105. doi:10.1101/pdb.emo105Koenig, D., Jimenez-Gomez, J. M., Kimura, S., Fulop, D., Chitwood, D. H., Headland, L. R., … Maloof, J. N. (2013). Comparative transcriptomics reveals patterns of selection in domesticated and wild tomato. Proceedings of the National Academy of Sciences, 110(28), E2655-E2662. doi:10.1073/pnas.1309606110Li, H., & Durbin, R. (2009). Fast and accurate short read alignment with Burrows-Wheeler transform. Bioinformatics, 25(14), 1754-1760. doi:10.1093/bioinformatics/btp324Li, H., Handsaker, B., Wysoker, A., Fennell, T., Ruan, J., … Homer, N. (2009). The Sequence Alignment/Map format and SAMtools. Bioinformatics, 25(16), 2078-2079. doi:10.1093/bioinformatics/btp352Li, J., Sima, W., Ouyang, B., Wang, T., Ziaf, K., Luo, Z., … Ye, Z. (2012). Tomato SlDREB gene restricts leaf expansion and internode elongation by downregulating key genes for gibberellin biosynthesis. Journal of Experimental Botany, 63(18), 6407-6420. doi:10.1093/jxb/ers295Lorrain, S., & Fankhauser, C. (2012). Plant Development: Should I Stop or Should I Grow? Current Biology, 22(16), R645-R647. doi:10.1016/j.cub.2012.06.054Menda, N., Semel, Y., Peled, D., Eshed, Y., & Zamir, D. (2004). In silicoscreening of a saturated mutation library of tomato. The Plant Journal, 38(5), 861-872. doi:10.1111/j.1365-313x.2004.02088.xMichelmore, R. W., Paran, I., & Kesseli, R. V. (1991). Identification of markers linked to disease-resistance genes by bulked segregant analysis: a rapid method to detect markers in specific genomic regions by using segregating populations. Proceedings of the National Academy of Sciences, 88(21), 9828-9832. doi:10.1073/pnas.88.21.9828Pimenta Lange, M. J., Liebrandt, A., Arnold, L., Chmielewska, S.-M., Felsberger, A., Freier, E., … Lange, T. (2013). Functional characterization of gibberellin oxidases from cucumber, Cucumis sativus L. Phytochemistry, 90, 62-69. doi:10.1016/j.phytochem.2013.02.006Raskin, I., & Kende, H. (1984). Role of Gibberellin in the Growth Response of Submerged Deep Water Rice. Plant Physiology, 76(4), 947-950. doi:10.1104/pp.76.4.947Reinecke, D. M., Wickramarathna, A. D., Ozga, J. A., Kurepin, L. V., Jin, A. L., Good, A. G., & Pharis, R. P. (2013). Gibberellin 3-oxidase Gene Expression Patterns Influence Gibberellin Biosynthesis, Growth, and Development in Pea. PLANT PHYSIOLOGY, 163(2), 929-945. doi:10.1104/pp.113.225987Robinson, M. D., & Oshlack, A. (2010). A scaling normalization method for differential expression analysis of RNA-seq data. Genome Biology, 11(3), R25. doi:10.1186/gb-2010-11-3-r25Robinson, M. D., McCarthy, D. J., & Smyth, G. K. (2009). edgeR: a Bioconductor package for differential expression analysis of digital gene expression data. Bioinformatics, 26(1), 139-140. doi:10.1093/bioinformatics/btp616Robinson, J. T., Thorvaldsdóttir, H., Winckler, W., Guttman, M., Lander, E. S., Getz, G., & Mesirov, J. P. (2011). Integrative genomics viewer. Nature Biotechnology, 29(1), 24-26. doi:10.1038/nbt.1754Schneeberger, K., Ossowski, S., Lanz, C., Juul, T., Petersen, A. H., Nielsen, K. L., … Andersen, S. U. (2009). SHOREmap: simultaneous mapping and mutation identification by deep sequencing. Nature Methods, 6(8), 550-551. doi:10.1038/nmeth0809-550Schneider, C. A., Rasband, W. S., & Eliceiri, K. W. (2012). NIH Image to ImageJ: 25 years of image analysis. Nature Methods, 9(7), 671-675. doi:10.1038/nmeth.2089Schomburg, F. M., Bizzell, C. M., Lee, D. J., Zeevaart, J. A. D., & Amasino, R. M. (2002). Overexpression of a Novel Class of Gibberellin 2-Oxidases Decreases Gibberellin Levels and Creates Dwarf Plants. The Plant Cell, 15(1), 151-163. doi:10.1105/tpc.005975Seo, M., Jikumaru, Y., & Kamiya, Y. (2011). Profiling of Hormones and Related Metabolites in Seed Dormancy and Germination Studies. Methods in Molecular Biology, 99-111. doi:10.1007/978-1-61779-231-1_7Sun, T. (2011). The Molecular Mechanism and Evolution of the GA–GID1–DELLA Signaling Module in Plants. Current Biology, 21(9), R338-R345. doi:10.1016/j.cub.2011.02.036Sun, T., & Gubler, F. (2004). MOLECULAR MECHANISM OF GIBBERELLIN SIGNALING IN PLANTS. Annual Review of Plant Biology, 55(1), 197-223. doi:10.1146/annurev.arplant.55.031903.141753Thorvaldsdottir, H., Robinson, J. T., & Mesirov, J. P. (2012). Integrative Genomics Viewer (IGV): high-performance genomics data visualization and exploration. Briefings in Bioinformatics, 14(2), 178-192. doi:10.1093/bib/bbs017(2012). The tomato genome sequence provides insights into fleshy fruit evolution. Nature, 485(7400), 635-641. doi:10.1038/nature11119Trapnell, C., Pachter, L., & Salzberg, S. L. (2009). TopHat: discovering splice junctions with RNA-Seq. Bioinformatics, 25(9), 1105-1111. doi:10.1093/bioinformatics/btp120Trapnell, C., Roberts, A., Goff, L., Pertea, G., Kim, D., Kelley, D. R., … Pachter, L. (2012). Differential gene and transcript expression analysis of RNA-seq experiments with TopHat and Cufflinks. Nature Protocols, 7(3), 562-578. doi:10.1038/nprot.2012.016Tsai, H., Howell, T., Nitcher, R., Missirian, V., Watson, B., Ngo, K. J., … Comai, L. (2011). Discovery of Rare Mutations in Populations: TILLING by Sequencing. Plant Physiology, 156(3), 1257-1268. doi:10.1104/pp.110.169748Ueguchi-Tanaka, M., Nakajima, M., Katoh, E., Ohmiya, H., Asano, K., Saji, S., … Matsuoka, M. (2007). Molecular Interactions of a Soluble Gibberellin Receptor, GID1, with a Rice DELLA Protein, SLR1, and Gibberellin. The Plant Cell, 19(7), 2140-2155. doi:10.1105/tpc.106.043729Wickham, H. (2016). ggplot2. Use R! doi:10.1007/978-3-319-24277-4Winter, D., Vinegar, B., Nahal, H., Ammar, R., Wilson, G. V., & Provart, N. J. (2007). An «Electronic Fluorescent Pictograph» Browser for Exploring and Analyzing Large-Scale Biological Data Sets. PLoS ONE, 2(8), e718. doi:10.1371/journal.pone.0000718Xu, H., Liu, Q., Yao, T., & Fu, X. (2014). Shedding light on integrative GA signaling. Current Opinion in Plant Biology, 21, 89-95. doi:10.1016/j.pbi.2014.06.010Yamaguchi, S. (2008). Gibberellin Metabolism and its Regulation. Annual Review of Plant Biology, 59(1), 225-251. doi:10.1146/annurev.arplant.59.032607.09280

Efeito radioprotetor do piruvato de etila, sozinho ou como um coadjuvante de amifostina

Entre los radioprotectores con uso clínico se destaca la amifostina (WR- 2721), eficaz pero con efectos secundarios que impiden su uso repetitivo. Es interés de los autores desarrollar radioprotectores menos tóxicos, por sí mismos o como coadyuvantes de amifostina. Ratas machos o hembras se expusieron a una dosis de rayos X de 2 Gy. Se ensayó el piruvato de etilo, solo o conjuntamente con amifostina. Cuarenta y ocho horas después de la exposición a la radiación, se realizó el recuento de eritrocitos, de leucocitos y la fórmula leucocitaria. Los efectos genotóxicos se evaluaron en leucocitos de sangre mediante el ensayo Cometa. Se realizaron también estudios de supervivencia a 60 días post-irradiación. En los animales irradiados disminuyeron los eritrocitos, y el recuento de leucocitos se redujo drásticamente respecto al control, presentando además una fórmula alterada. El tratamiento con piruvato de etilo resultó en una protección de los eritrocitos en ambos sexos. El daño genético disminuyó significativamente por el tratamiento con piruvato de etilo solo o combinado con amifostina, y en hembras se observó una mayor supervivencia solo con el tratamiento combinado. El piruvato de etilo mostró una acción radioprotectora significativa, que podría mejorarse aumentando la dosis o el tiempo de tratamiento, ya que tiene muy baja toxicidad.Among the currently available radioprotectors, only amifostine (WR-2721) has shown in clinical trials to reduce radiation-induced toxicity. This compound is an efficient radioprotector but it exhibits some undesirable side effects which prevent its repetitive use. Efforts are directed to develop radioprotective agents with lower toxicity, with their own protective potential or suitable as coadyuvants of amifostine. The present study describes the results obtained by repetitive oral administration of ethyl pyruvate. Male or female rats were exposed to an X-ray dose of 2 Gy. Forty-eight hours after exposure to radiation, erythrocyte count, leukocyte and differential count were performed. Genotoxic effects were assessed in blood leukocytes by the Comet assay. Survival studies were also performed at 60 days post-irradiation. Eritrocyte and leukocyte were reduced in animals exposed to radiation compared to the control, also presenting an altered formula. Treatment with ethyl pyruvate resulted in a protection on erythrocytes of both sexes. Genetic damage was significantly decreased by ethyl pyruvate alone or combined with amifostine, and in females, higher survival was observed only with combined administration. Ethyl pyruvate showed a significant radioprotective action, which could be improved by increasing the dose or time of treatment because it has low toxicity.Entre os radioprotetores com uso clínico destaca-se a amifostina (WR-2721) eficaz mas com efeitos secundários que impedem seu uso repetitivo. O interesse dos autores é desenvolver radioprotetores menos tóxicos, por si mesmos ou como coadjuvantes de amistofina. Ratos machos ou fêmeas foram expostos a doses de raios X de 2Gy. Ensaiou-se o piruvato de etila, só ou junto com amifostina. Quarenta e oito horas após a exposição à radiação foi realizada a contagem de eritrócitos, de leucócitos e da fórmula leucocitária. Efeitos genotóxicos foram avaliados em leucócitos do sangue pelo Ensaio Cometa. Estudos de sobrevivência foram também realizados a 60 dias pós-irradiação. Nos animais irradiados diminuíram os eritrócitos, e a contagem de leucócitos se reduziu drasticamente em comparação com o controle, apresentando também uma fórmula alterada. O tratamento com piruvato de etila resultou numa proteção dos eritrócitos em ambos os sexos. O dano genético diminuiu significativamente pelo tratamento com piruvato de etila sozinho ou combinado com amifostina, e nas fêmeas se observou maior sobrevivência só com o tratamento combinado. O piruvato de etila mostrou uma ação radioprotetora significativa, que poderia ser melhorada pelo aumento da dose ou do tempo de tratamento, visto que tem baixa toxicidade.Fil: Maciel, Maria Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; ArgentinaFil: Quintans, Leandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; ArgentinaFil: Diaz Gomez, Maria Isabel. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; ArgentinaFil: Costantini, Martin Hernan. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; ArgentinaFil: Formosa Lemoine, Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; ArgentinaFil: Montalto, Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; ArgentinaFil: Lopez, Gabriel Diego. Ministerio de Defensa. Instituto de Investigaciones Científicas y Técnicas para la Defensa; ArgentinaFil: Castro, Jose Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; ArgentinaFil: Castro, Gerardo Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Unidad de Investigación y Desarrollo Estratégico para la Defensa. Ministerio de Defensa. Unidad de Investigación y Desarrollo Estratégico para la Defensa; Argentina. Universidad Nacional de San Martín. Instituto de Investigación e Ingeniería Ambiental; Argentin

Human biomonitoring. Basics: educational course

Ana Isabel Cañas Portilla, Argelia Castaño, Susana Pedraza Diaz y Marta Esteban López del Centro Nacional de Sanidad Ambiental (ISCIII) han contribuido en el desarrollo de este curso.Human biomonitoring (HBM) is an instrument for measuring the internal dose of exogenous substances/chemicals that enter a body during a certain period of exposure from a range of sources. It contributes to reducing uncertainties in the assessment of health risks from chemicals and provides information for decision-making on the prevention of negative impacts of chemicals on human health and the environment. Promoting the use of HBM is a recognized priority of chemical safety globally and in the WHO European Region. Given the complexity of HBM, relevant capacities should be built at the national level to explore its benefits. This educational course on HBM, presented in the form of slides with accompanying notes and references, compiles scientific information on HBM as well as practical examples. It was developed to support the training of public-health and health-care professionals; students of medical, biological and other allied sciences; and professionals and decision-makers in the health, environment and other relevant sectorsThis course was developed with financial support from the German Federal Ministry for the Environment, Nature Conservation, Nuclear Safety and Consumer Protection and the German Federal Ministry of Health.S

Regulation of ovule initiation by gibberellins and brassinosteroids in tomato and Arabidopsis: two plant species, two molecular mechanisms

This is the peer reviewed version of the following article: Barro¿Trastoy, D., Carrera, E., Baños, J., Palau-Rodríguez, J., Ruiz-Rivero, O., Tornero, P., Alonso, J.M., López-Díaz, I., Gómez, M.D. and Pérez-Amador, M.A. (2020), Regulation of ovule initiation by gibberellins and brassinosteroids in tomato and Arabidopsis: two plant species, two molecular mechanisms. Plant J, 102: 1026-1041, which has been published in final form at https://doi.org/10.1111/tpj.14684. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.[EN] Ovule primordia formation is a complex developmental process with a strong impact on the production of seeds. In Arabidopsis this process is controlled by a gene network, including components of the signalling pathways of auxin, brassinosteroids (BRs) and cytokinins. Recently, we have shown that gibberellins (GAs) also play an important role in ovule primordia initiation, inhibiting ovule formation in both Arabidopsis and tomato. Here we reveal that BRs also participate in the control of ovule initiation in tomato, by promoting an increase on ovule primordia formation. Moreover, molecular and genetic analyses of the co-regulation by GAs and BRs of the control of ovule initiation indicate that two different mechanisms occur in tomato and Arabidopsis. In tomato, GAs act downstream of BRs. BRs regulate ovule number through the downregulation of GA biosynthesis, which provokes stabilization of DELLA proteins that will finally promote ovule primordia initiation. In contrast, in Arabidopsis both GAs and BRs regulate ovule number independently of the activity levels of the other hormone. Taken together, our data strongly suggest that different molecular mechanisms could operate in different plant species to regulate identical developmental processes even, as for ovule primordia initiation, if the same set of hormones trigger similar responses, adding a new level of complexity.We wish to thank B. Janssen (Horticulture and Food Research Institute, New Zealand) for the pBJ60 shuttle vector, C. Ferrandiz and M. Colombo (IBMCP, CSIC-UPV, Valencia, Spain) for their help in the generation of 35S:ANT lines and L.E.P. Peres (Universidade de Sao Paulo, Brazil) for the tomato mutant lines. Our thanks also go to C. Fuster for technical assistance. This work was supported by grants from the Spanish Ministry of Economy and Competitiveness-FEDER (BIO2017-83138R) to MAPA and from NSF (DBI-0820755, MCB-1158181, and IOS-1444561) to JMA.Barro-Trastoy, D.; Carrera, E.; Baños, J.; Palau-Rodríguez, J.; Ruiz-Rivero, O.; Tornero Feliciano, P.; Alonso, JM.... (2020). Regulation of ovule initiation by gibberellins and brassinosteroids in tomato and Arabidopsis: two plant species, two molecular mechanisms. The Plant Journal. 102(5):1026-1041. https://doi.org/10.1111/tpj.14684S102610411025Azhakanandam, S., Nole-Wilson, S., Bao, F., & Franks, R. G. (2008). SEUSSandAINTEGUMENTAMediate Patterning and Ovule Initiation during Gynoecium Medial Domain Development    . Plant Physiology, 146(3), 1165-1181. doi:10.1104/pp.107.114751Bai, M.-Y., Shang, J.-X., Oh, E., Fan, M., Bai, Y., Zentella, R., … Wang, Z.-Y. (2012). Brassinosteroid, gibberellin and phytochrome impinge on a common transcription module in Arabidopsis. Nature Cell Biology, 14(8), 810-817. doi:10.1038/ncb2546Baker, S. C., Robinson-Beers, K., Villanueva, J. M., Gaiser, J. C., & Gasser, C. S. (1997). Interactions Among Genes Regulating Ovule Development in Arabidopsis thaliana. Genetics, 145(4), 1109-1124. doi:10.1093/genetics/145.4.1109Bartrina, I., Otto, E., Strnad, M., Werner, T., & Schmülling, T. (2011). Cytokinin Regulates the Activity of Reproductive Meristems, Flower Organ Size, Ovule Formation, and Thus Seed Yield in Arabidopsis thaliana      . The Plant Cell, 23(1), 69-80. doi:10.1105/tpc.110.079079Belkhadir, Y., & Jaillais, Y. (2015). The molecular circuitry of brassinosteroid signaling. New Phytologist, 206(2), 522-540. doi:10.1111/nph.13269Bencivenga, S., Simonini, S., Benková, E., & Colombo, L. (2012). The Transcription Factors BEL1 and SPL Are Required for Cytokinin and Auxin Signaling During Ovule Development in Arabidopsis. The Plant Cell, 24(7), 2886-2897. doi:10.1105/tpc.112.100164Brumos, J., Zhao, C., Gong, Y., Soriano, D., Patel, A. P., Perez-Amador, M. A., … Alonso, J. M. (2019). An Improved Recombineering Toolset for Plants. The Plant Cell, 32(1), 100-122. doi:10.1105/tpc.19.00431Carrera, E., Ruiz-Rivero, O., Peres, L. E. P., Atares, A., & Garcia-Martinez, J. L. (2012). Characterization of the procera Tomato Mutant Shows Novel Functions of the SlDELLA Protein in the Control of Flower Morphology, Cell Division and Expansion, and the Auxin-Signaling Pathway during Fruit-Set and Development    . Plant Physiology, 160(3), 1581-1596. doi:10.1104/pp.112.204552Carvalho, R. F., Campos, M. L., Pino, L. E., Crestana, S. L., Zsögön, A., Lima, J. E., … Peres, L. E. (2011). Convergence of developmental mutants into a single tomato model system: «Micro-Tom» as an effective toolkit for plant development research. Plant Methods, 7(1). doi:10.1186/1746-4811-7-18Chory, J., Nagpal, P., & Peto, C. A. (1991). Phenotypic and Genetic Analysis of det2, a New Mutant That Affects Light-Regulated Seedling Development in Arabidopsis. The Plant Cell, 3(5), 445. doi:10.2307/3869351Clough, S. J., & Bent, A. F. (1998). Floral dip: a simplified method forAgrobacterium-mediated transformation ofArabidopsis thaliana. The Plant Journal, 16(6), 735-743. doi:10.1046/j.1365-313x.1998.00343.xClouse, S. D. (2011). Brassinosteroid Signal Transduction: From Receptor Kinase Activation to Transcriptional Networks Regulating Plant Development. The Plant Cell, 23(4), 1219-1230. doi:10.1105/tpc.111.084475Cucinotta, M., Colombo, L., & Roig-Villanova, I. (2014). Ovule development, a new model for lateral organ formation. Frontiers in Plant Science, 5. doi:10.3389/fpls.2014.00117Cucinotta, M., Manrique, S., Guazzotti, A., Quadrelli, N. E., Mendes, M. A., Benkova, E., & Colombo, L. (2016). Cytokinin response factors integrate auxin and cytokinin pathways for female reproductive organ development. Development. doi:10.1242/dev.143545Davière, J.-M., & Achard, P. (2016). A Pivotal Role of DELLAs in Regulating Multiple Hormone Signals. Molecular Plant, 9(1), 10-20. doi:10.1016/j.molp.2015.09.011De Vleesschauwer, D., Van Buyten, E., Satoh, K., Balidion, J., Mauleon, R., Choi, I.-R., … Höfte, M. (2012). Brassinosteroids Antagonize Gibberellin- and Salicylate-Mediated Root Immunity in Rice      . Plant Physiology, 158(4), 1833-1846. doi:10.1104/pp.112.193672Dorcey, E., Urbez, C., Blázquez, M. A., Carbonell, J., & Perez-Amador, M. A. (2009). Fertilization-dependent auxin response in ovules triggers fruit development through the modulation of gibberellin metabolism in Arabidopsis. The Plant Journal, 58(2), 318-332. doi:10.1111/j.1365-313x.2008.03781.xFujioka, S., Li, J., Choi, Y. H., Seto, H., Takatsuto, S., Noguchi, T., … Sakurai, A. (1997). The Arabidopsis deetiolated2 mutant is blocked early in brassinosteroid biosynthesis. The Plant Cell, 9(11), 1951-1962. doi:10.1105/tpc.9.11.1951Galbiati, F., Sinha Roy, D., Simonini, S., Cucinotta, M., Ceccato, L., Cuesta, C., … Colombo, L. (2013). An integrative model of the control of ovule primordia formation. The Plant Journal, 76(3), 446-455. doi:10.1111/tpj.12309Gallego-Bartolome, J., Minguet, E. G., Grau-Enguix, F., Abbas, M., Locascio, A., Thomas, S. G., … Blazquez, M. A. (2012). Molecular mechanism for the interaction between gibberellin and brassinosteroid signaling pathways in Arabidopsis. Proceedings of the National Academy of Sciences, 109(33), 13446-13451. doi:10.1073/pnas.1119992109García-Hurtado, N., Carrera, E., Ruiz-Rivero, O., López-Gresa, M. P., Hedden, P., Gong, F., & García-Martínez, J. L. (2012). The characterization of transgenic tomato overexpressing gibberellin 20-oxidase reveals induction of parthenocarpic fruit growth, higher yield, and alteration of the gibberellin biosynthetic pathway. Journal of Experimental Botany, 63(16), 5803-5813. doi:10.1093/jxb/ers229Gleave, A. P. (1992). A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome. Plant Molecular Biology, 20(6), 1203-1207. doi:10.1007/bf00028910Gomez, M. D., Ventimilla, D., Sacristan, R., & Perez-Amador, M. A. (2016). Gibberellins Regulate Ovule Integument Development by Interfering with the Transcription Factor ATS. Plant Physiology, 172(4), 2403-2415. doi:10.1104/pp.16.01231He, J.-X., Gendron, J. M., Sun, Y., Gampala, S. S. L., Gendron, N., Sun, C. Q., & Wang, Z.-Y. (2005). BZR1 Is a Transcriptional Repressor with Dual Roles in Brassinosteroid Homeostasis and Growth Responses. Science, 307(5715), 1634-1638. doi:10.1126/science.1107580Huang, H.-Y., Jiang, W.-B., Hu, Y.-W., Wu, P., Zhu, J.-Y., Liang, W.-Q., … Lin, W.-H. (2013). BR Signal Influences Arabidopsis Ovule and Seed Number through Regulating Related Genes Expression by BZR1. Molecular Plant, 6(2), 456-469. doi:10.1093/mp/sss070Kurepin, L. V., Joo, S.-H., Kim, S.-K., Pharis, R. P., & Back, T. G. (2011). Interaction of Brassinosteroids with Light Quality and Plant Hormones in Regulating Shoot Growth of Young Sunflower and Arabidopsis Seedlings. Journal of Plant Growth Regulation, 31(2), 156-164. doi:10.1007/s00344-011-9227-7Li, Q.-F., Wang, C., Jiang, L., Li, S., Sun, S. S. M., & He, J.-X. (2012). An Interaction Between BZR1 and DELLAs Mediates Direct Signaling Crosstalk Between Brassinosteroids and Gibberellins in Arabidopsis. Science Signaling, 5(244). doi:10.1126/scisignal.2002908Li, X.-J., Chen, X.-J., Guo, X., Yin, L.-L., Ahammed, G. J., Xu, C.-J., … Yu, J.-Q. (2015). DWARFoverexpression induces alteration in phytohormone homeostasis, development, architecture and carotenoid accumulation in tomato. Plant Biotechnology Journal, 14(3), 1021-1033. doi:10.1111/pbi.12474Liu, Z., Franks, R. G., & Klink, V. P. (2000). Regulation of Gynoecium Marginal Tissue Formation by LEUNIG and AINTEGUMENTA. The Plant Cell, 12(10), 1879-1891. doi:10.1105/tpc.12.10.1879Marti, E. (2006). Genetic and physiological characterization of tomato cv. Micro-Tom. Journal of Experimental Botany, 57(9), 2037-2047. doi:10.1093/jxb/erj154Mizukami, Y., & Fischer, R. L. (2000). Plant organ size control: AINTEGUMENTA regulates growth and cell numbers during organogenesis. Proceedings of the National Academy of Sciences, 97(2), 942-947. doi:10.1073/pnas.97.2.942Montoya, T., Nomura, T., Yokota, T., Farrar, K., Harrison, K., Jones, J. G. D., … Bishop, G. J. (2005). Patterns of Dwarf expression and brassinosteroid accumulation in tomato reveal the importance of brassinosteroid synthesis during fruit development. The Plant Journal, 42(2), 262-269. doi:10.1111/j.1365-313x.2005.02376.xMüller, C. J., Larsson, E., Spíchal, L., & Sundberg, E. (2017). Cytokinin-Auxin Crosstalk in the Gynoecial Primordium Ensures Correct Domain Patterning. Plant Physiology, 175(3), 1144-1157. doi:10.1104/pp.17.00805Murashige, T., & Skoog, F. (1962). A Revised Medium for Rapid Growth and Bio Assays with Tobacco Tissue Cultures. Physiologia Plantarum, 15(3), 473-497. doi:10.1111/j.1399-3054.1962.tb08052.xOlimpieri, I., Siligato, F., Caccia, R., Soressi, G. P., Mazzucato, A., Mariotti, L., & Ceccarelli, N. (2007). Tomato fruit set driven by pollination or by the parthenocarpic fruit allele are mediated by transcriptionally regulated gibberellin biosynthesis. Planta, 226(4), 877-888. doi:10.1007/s00425-007-0533-zPaz-Ares, J., & The REGIA Consortium. (2002). REGIA, An EU Project on Functional Genomics of Transcription Factors fromArabidopsis thaliana. Comparative and Functional Genomics, 3(2), 102-108. doi:10.1002/cfg.146Peng, J., Carol, P., Richards, D. E., King, K. E., Cowling, R. J., Murphy, G. P., & Harberd, N. P. (1997). The Arabidopsis GAI gene defines a signaling pathway that negatively regulates gibberellin responses . Genes & Development, 11(23), 3194-3205. doi:10.1101/gad.11.23.3194Reyes-Olalde, J. I., Zuñiga-Mayo, V. M., Chávez Montes, R. A., Marsch-Martínez, N., & de Folter, S. (2013). Inside the gynoecium: at the carpel margin. Trends in Plant Science, 18(11), 644-655. doi:10.1016/j.tplants.2013.08.002Sabelli, P. A., & Larkins, B. A. (2009). The Development of Endosperm in Grasses. Plant Physiology, 149(1), 14-26. doi:10.1104/pp.108.129437Schneitz, K., Baker, S. C., Gasser, C. S., & Redweik, A. (1998). Pattern formation and growth during floral organogenesis: HUELLENLOS and AINTEGUMENTA are required for the formation of the proximal region of the ovule primordium in Arabidopsis thaliana. Development, 125(14), 2555-2563. doi:10.1242/dev.125.14.2555Schneitz, K., Hulskamp, M., & Pruitt, R. E. (1995). Wild-type ovule development in Arabidopsis thaliana: a light microscope study of cleared whole-mount tissue. The Plant Journal, 7(5), 731-749. doi:10.1046/j.1365-313x.1995.07050731.xSeo, M., Jikumaru, Y., & Kamiya, Y. (2011). Profiling of Hormones and Related Metabolites in Seed Dormancy and Germination Studies. Methods in Molecular Biology, 99-111. doi:10.1007/978-1-61779-231-1_7Serrani, J. C., Sanjuán, R., Ruiz-Rivero, O., Fos, M., & García-Martínez, J. L. (2007). Gibberellin Regulation of Fruit Set and Growth in Tomato. Plant Physiology, 145(1), 246-257. doi:10.1104/pp.107.098335Serrani, J. C., Carrera, E., Ruiz-Rivero, O., Gallego-Giraldo, L., Peres, Lá. E. P., & García-Martínez, J. L. (2010). Inhibition of Auxin Transport from the Ovary or from the Apical Shoot Induces Parthenocarpic Fruit-Set in Tomato Mediated by Gibberellins    . Plant Physiology, 153(2), 851-862. doi:10.1104/pp.110.155424Sun, T. (2010). Gibberellin-GID1-DELLA: A Pivotal Regulatory Module for Plant Growth and Development. Plant Physiology, 154(2), 567-570. doi:10.1104/pp.110.161554Sun, T. (2011). The Molecular Mechanism and Evolution of the GA–GID1–DELLA Signaling Module in Plants. Current Biology, 21(9), R338-R345. doi:10.1016/j.cub.2011.02.036Tanaka, K., Nakamura, Y., Asami, T., Yoshida, S., Matsuo, T., & Okamoto, S. (2003). Physiological Roles of Brassinosteroids in Early Growth of Arabidopsis: Brassinosteroids Have a Synergistic Relationship with Gibberellin as well as Auxin in Light-Grown Hypocotyl Elongation. Journal of Plant Growth Regulation, 22(3), 259-271. doi:10.1007/s00344-003-0119-3Tang, Y., Liu, H., Guo, S., Wang, B., Li, Z., Chong, K., & Xu, Y. (2017). OsmiR396d Affects Gibberellin and Brassinosteroid Signaling to Regulate Plant Architecture in Rice. Plant Physiology, 176(1), 946-959. doi:10.1104/pp.17.00964Tong, H., Xiao, Y., Liu, D., Gao, S., Liu, L., Yin, Y., … Chu, C. (2014). Brassinosteroid Regulates Cell Elongation by Modulating Gibberellin Metabolism in Rice    . The Plant Cell, 26(11), 4376-4393. doi:10.1105/tpc.114.132092Truernit, E., Bauby, H., Dubreucq, B., Grandjean, O., Runions, J., Barthélémy, J., & Palauqui, J.-C. (2008). High-Resolution Whole-Mount Imaging of Three-Dimensional Tissue Organization and Gene Expression Enables the Study of Phloem Development and Structure inArabidopsis . The Plant Cell, 20(6), 1494-1503. doi:10.1105/tpc.107.056069Tursun, B., Cochella, L., Carrera, I., & Hobert, O. (2009). A Toolkit and Robust Pipeline for the Generation of Fosmid-Based Reporter Genes in C. elegans. PLoS ONE, 4(3), e4625. doi:10.1371/journal.pone.0004625Unterholzner, S. J., Rozhon, W., Papacek, M., Ciomas, J., Lange, T., Kugler, K. G., … Poppenberger, B. (2015). Brassinosteroids Are Master Regulators of Gibberellin Biosynthesis in Arabidopsis. The Plant Cell, 27(8), 2261-2272. doi:10.1105/tpc.15.00433Wang, Z.-Y., Nakano, T., Gendron, J., He, J., Chen, M., Vafeados, D., … Chory, J. (2002). Nuclear-Localized BZR1 Mediates Brassinosteroid-Induced Growth and Feedback Suppression of Brassinosteroid Biosynthesis. Developmental Cell, 2(4), 505-513. doi:10.1016/s1534-5807(02)00153-3Xiao, H., Radovich, C., Welty, N., Hsu, J., Li, D., Meulia, T., & van der Knaap, E. (2009). Integration of tomato reproductive developmental landmarks and expression profiles, and the effect of SUN on fruit shape. BMC Plant Biology, 9(1). doi:10.1186/1471-2229-9-49Xiao, Y., Liu, D., Zhang, G., Tong, H., & Chu, C. (2017). Brassinosteroids Regulate OFP1, a DLT Interacting Protein, to Modulate Plant Architecture and Grain Morphology in Rice. Frontiers in Plant Science, 8. doi:10.3389/fpls.2017.0169

Tomato floral induction and flower development are orchestrated by the interplay between gibberellin and two unrelated microRNA-controlled modules

[EN] Age-regulated microRNA156 (miR156) and targets similarly control the competence to flower in diverse species. By contrast, the diterpene hormone gibberellin (GA) and the microRNA319-regulated TEOSINTE BRANCHED/CYCLOIDEA/PCF (TCP) transcription factors promote flowering in the facultative long-day Arabidopsis thaliana, but suppress it in the day-neutral tomato (Solanum lycopersicum). We combined genetic and molecular studies and described a new interplay between GA and two unrelated miRNA-associated pathways that modulates tomato transition to flowering. Tomato PROCERA/DELLA activity is required to promote flowering along with the miR156-targeted SQUAMOSA PROMOTER BINDING-LIKE (SPL/SBP) transcription factors by activating SINGLE FLOWER TRUSS (SFT) in the leaves and the MADS-Boxgene APETALA1(AP1)/MC at the shoot apex. Conversely, miR319-targeted LANCEOLATE represses floral transition by increasing GA concentrations and inactivating SFT in the leaves and AP1/MC at the shoot apex. Importantly, the combination of high GA concentrations/responses with the loss of SPL/SPB function impaired canonical meristem maturation and flower initiation in tomato. Our results reveal a cooperative regulation of tomato floral induction and flower development, integrating age cues (miR156 module) with GA responses and miR319-controlled pathways. Importantly, this study contributes to elucidate the mechanisms underlying the effects of GA in controlling flowering time in a day-neutral species.We thank Dr C. Schommer for kindly providing tcp4-soj8/+ seeds, and Carlos Rojas for Arabidopsis flowering time analyses. This work was supported by FAPESP (grant no. 15/17892-7 and fellowships nos 15/23826-7 and 13/16949-0). The authors declare no conflict of interest.Silva, G.; Silva, E.; Correa, J.; Vicente, M.; Jiang, N.; Notini, M.; Junior, A.... (2018). Tomato floral induction and flower development are orchestrated by the interplay between gibberellin and two unrelated microRNA-controlled modules. New Phytologist. 221(3):1328-1344. https://doi.org/10.1111/nph.15492S132813442213Andrés, F., & Coupland, G. (2012). The genetic basis of flowering responses to seasonal cues. Nature Reviews Genetics, 13(9), 627-639. doi:10.1038/nrg3291Bassel, G. W., Mullen, R. T., & Bewley, J. D. (2008). procerais a putative DELLA mutant in tomato (Solanum lycopersicum): effects on the seed and vegetative plant. Journal of Experimental Botany, 59(3), 585-593. doi:10.1093/jxb/erm354Ben‐Naim, O., Eshed, R., Parnis, A., Teper‐Bamnolker, P., Shalit, A., Coupland, G., … Lifschitz, E. (2006). The CCAAT binding factor can mediate interactions between CONSTANS‐like proteins and DNA. The Plant Journal, 46(3), 462-476. doi:10.1111/j.1365-313x.2006.02706.xBoss, P. K., & Thomas, M. R. (2002). Association of dwarfism and floral induction with a grape ‘green revolution’ mutation. Nature, 416(6883), 847-850. doi:10.1038/416847aBurko, Y., Shleizer-Burko, S., Yanai, O., Shwartz, I., Zelnik, I. D., Jacob-Hirsch, J., … Ori, N. (2013). A Role for APETALA1/FRUITFULL Transcription Factors in Tomato Leaf Development. The Plant Cell, 25(6), 2070-2083. doi:10.1105/tpc.113.113035Cardon, G., Höhmann, S., Klein, J., Nettesheim, K., Saedler, H., & Huijser, P. (1999). Molecular characterisation of the Arabidopsis SBP-box genes. Gene, 237(1), 91-104. doi:10.1016/s0378-1119(99)00308-xCarrera, E., Ruiz-Rivero, O., Peres, L. E. P., Atares, A., & Garcia-Martinez, J. L. (2012). Characterization of the procera Tomato Mutant Shows Novel Functions of the SlDELLA Protein in the Control of Flower Morphology, Cell Division and Expansion, and the Auxin-Signaling Pathway during Fruit-Set and Development. Plant Physiology, 160(3), 1581-1596. doi:10.1104/pp.112.204552Carvalho, R. F., Campos, M. L., Pino, L. E., Crestana, S. L., Zsögön, A., Lima, J. E., … Peres, L. E. (2011). Convergence of developmental mutants into a single tomato model system: «Micro-Tom» as an effective toolkit for plant development research. Plant Methods, 7(1), 18. doi:10.1186/1746-4811-7-18Cubas, P., Lauter, N., Doebley, J., & Coen, E. (1999). The TCP domain: a motif found in proteins regulating plant growth and development. The Plant Journal, 18(2), 215-222. doi:10.1046/j.1365-313x.1999.00444.xDavière, J.-M., Wild, M., Regnault, T., Baumberger, N., Eisler, H., Genschik, P., & Achard, P. (2014). Class I TCP-DELLA Interactions in Inflorescence Shoot Apex Determine Plant Height. Current Biology, 24(16), 1923-1928. doi:10.1016/j.cub.2014.07.012Silva, G. F. F. e, Silva, E. M., da Silva Azevedo, M., Guivin, M. A. C., Ramiro, D. A., Figueiredo, C. R., … Nogueira, F. T. S. (2014). microRNA156-targeted SPL/SBP box transcription factors regulate tomato ovary and fruit development. The Plant Journal, 78(4), 604-618. doi:10.1111/tpj.12493Gallego-Bartolome, J., Minguet, E. G., Marin, J. A., Prat, S., Blazquez, M. A., & Alabadi, D. (2010). Transcriptional Diversification and Functional Conservation between DELLA Proteins in Arabidopsis. Molecular Biology and Evolution, 27(6), 1247-1256. doi:10.1093/molbev/msq012Gallego-Giraldo, L., García-Martínez, J. L., Moritz, T., & López-Díaz, I. (2007). Flowering in Tobacco Needs Gibberellins but is not Promoted by the Levels of Active GA1 and GA4 in the Apical Shoot. Plant and Cell Physiology, 48(4), 615-625. doi:10.1093/pcp/pcm034Galvao, V. C., Horrer, D., Kuttner, F., & Schmid, M. (2012). Spatial control of flowering by DELLA proteins in Arabidopsis thaliana. Development, 139(21), 4072-4082. doi:10.1242/dev.080879García-Hurtado, N., Carrera, E., Ruiz-Rivero, O., López-Gresa, M. P., Hedden, P., Gong, F., & García-Martínez, J. L. (2012). The characterization of transgenic tomato overexpressing gibberellin 20-oxidase reveals induction of parthenocarpic fruit growth, higher yield, and alteration of the gibberellin biosynthetic pathway. Journal of Experimental Botany, 63(16), 5803-5813. doi:10.1093/jxb/ers229Gargul, J. M., Mibus, H., & Serek, M. (2013). Constitutive overexpression of Nicotiana GA 2 ox leads to compact phenotypes and delayed flowering in Kalanchoë blossfeldiana and Petunia hybrida. Plant Cell, Tissue and Organ Culture (PCTOC), 115(3), 407-418. doi:10.1007/s11240-013-0372-5Goldberg-Moeller, R., Shalom, L., Shlizerman, L., Samuels, S., Zur, N., Ophir, R., … Sadka, A. (2013). Effects of gibberellin treatment during flowering induction period on global gene expression and the transcription of flowering-control genes in Citrus buds. Plant Science, 198, 46-57. doi:10.1016/j.plantsci.2012.09.012Hauvermale, A. L., Ariizumi, T., & Steber, C. M. (2012). Gibberellin Signaling: A Theme and Variations on DELLA Repression. Plant Physiology, 160(1), 83-92. doi:10.1104/pp.112.200956Hyun, Y., Richter, R., Vincent, C., Martinez-Gallegos, R., Porri, A., & Coupland, G. (2016). Multi-layered Regulation of SPL15 and Cooperation with SOC1 Integrate Endogenous Flowering Pathways at the Arabidopsis Shoot Meristem. Developmental Cell, 37(3), 254-266. doi:10.1016/j.devcel.2016.04.001Itoh, H., Ueguchi-Tanaka, M., Sato, Y., Ashikari, M., & Matsuoka, M. (2002). The Gibberellin Signaling Pathway Is Regulated by the Appearance and Disappearance of SLENDER RICE1 in Nuclei. The Plant Cell, 14(1), 57-70. doi:10.1105/tpc.010319Jung, J.-H., Ju, Y., Seo, P. J., Lee, J.-H., & Park, C.-M. (2011). The SOC1-SPL module integrates photoperiod and gibberellic acid signals to control flowering time in Arabidopsis. The Plant Journal, 69(4), 577-588. doi:10.1111/j.1365-313x.2011.04813.xKing, R. W., & Ben-Tal, Y. (2001). A Florigenic Effect of Sucrose in Fuchsia hybrida Is Blocked by Gibberellin-Induced Assimilate Competition. Plant Physiology, 125(1), 488-496. doi:10.1104/pp.125.1.488Kubota, A., Ito, S., Shim, J. S., Johnson, R. S., Song, Y. H., Breton, G., … Imaizumi, T. (2017). TCP4-dependent induction of CONSTANS transcription requires GIGANTEA in photoperiodic flowering in Arabidopsis. PLOS Genetics, 13(6), e1006856. doi:10.1371/journal.pgen.1006856Kudla, J., & Bock, R. (2016). Lighting the Way to Protein-Protein Interactions: Recommendations on Best Practices for Bimolecular Fluorescence Complementation Analyses. The Plant Cell, 28(5), 1002-1008. doi:10.1105/tpc.16.00043Lifschitz, E., Eviatar, T., Rozman, A., Shalit, A., Goldshmidt, A., Amsellem, Z., … Eshed, Y. (2006). The tomato FT ortholog triggers systemic signals that regulate growth and flowering and substitute for diverse environmental stimuli. Proceedings of the National Academy of Sciences, 103(16), 6398-6403. doi:10.1073/pnas.0601620103Liu, J., Cheng, X., Liu, P., Li, D., Chen, T., Gu, X., & Sun, J. (2017). MicroRNA319-regulated TCPs interact with FBHs and PFT1 to activate CO transcription and control flowering time in Arabidopsis. PLOS Genetics, 13(5), e1006833. doi:10.1371/journal.pgen.1006833Livak, K. J., & Schmittgen, T. D. (2001). Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT Method. Methods, 25(4), 402-408. doi:10.1006/meth.2001.1262Livne, S., Lor, V. S., Nir, I., Eliaz, N., Aharoni, A., Olszewski, N. E., … Weiss, D. (2015). Uncovering DELLA-Independent Gibberellin Responses by Characterizing New Tomato procera Mutants. The Plant Cell, 27(6), 1579-1594. doi:10.1105/tpc.114.132795Lombardi-Crestana, S., da Silva Azevedo, M., e Silva, G. F. F., Pino, L. E., Appezzato-da-Glória, B., Figueira, A., … Peres, L. E. P. (2012). The Tomato (Solanum Lycopersicum cv. Micro-Tom) Natural Genetic Variation Rg1 and the DELLA Mutant Procera Control the Competence Necessary to Form Adventitious Roots and Shoots. Journal of Experimental Botany, 63(15), 5689-5703. doi:10.1093/jxb/ers221Lozano, R., Gimenez, E., Cara, B., Capel, J., & Angosto, T. (2009). Genetic analysis of reproductive development in tomato. The International Journal of Developmental Biology, 53(8-9-10), 1635-1648. doi:10.1387/ijdb.072440rlMartin, K., Kopperud, K., Chakrabarty, R., Banerjee, R., Brooks, R., & Goodin, M. M. (2009). Transient expression inNicotiana benthamianafluorescent marker lines provides enhanced definition of protein localization, movement and interactionsin planta. The Plant Journal, 59(1), 150-162. doi:10.1111/j.1365-313x.2009.03850.xMartínez-Bello, L., Moritz, T., & López-Díaz, I. (2015). Silencing C19-GA 2-oxidases induces parthenocarpic development and inhibits lateral branching in tomato plants. Journal of Experimental Botany, 66(19), 5897-5910. doi:10.1093/jxb/erv300Meissner, R., Chague, V., Zhu, Q., Emmanuel, E., Elkind, Y., & Levy, A. A. (2000). A high throughput system for transposon tagging and promoter trapping in tomato. The Plant Journal, 22(3), 265-274. doi:10.1046/j.1365-313x.2000.00735.xMolinero-Rosales, N., Jamilena, M., Zurita, S., Gomez, P., Capel, J., & Lozano, R. (1999). FALSIFLORA, the tomato orthologue of FLORICAULA and LEAFY, controls flowering time and floral meristem identity. The Plant Journal, 20(6), 685-693. doi:10.1046/j.1365-313x.1999.00641.xMorea, E. G. O., da Silva, E. M., e Silva, G. F. F., Valente, G. T., Barrera Rojas, C. H., Vincentz, M., & Nogueira, F. T. S. (2016). Functional and evolutionary analyses of the miR156 and miR529 families in land plants. BMC Plant Biology, 16(1). doi:10.1186/s12870-016-0716-5Mounet, F., Moing, A., Garcia, V., Petit, J., Maucourt, M., Deborde, C., … Lemaire-Chamley, M. (2009). Gene and Metabolite Regulatory Network Analysis of Early Developing Fruit Tissues Highlights New Candidate Genes for the Control of Tomato Fruit Composition and Development. Plant Physiology, 149(3), 1505-1528. doi:10.1104/pp.108.133967Nir, I., Shohat, H., Panizel, I., Olszewski, N., Aharoni, A., & Weiss, D. (2017). The Tomato DELLA Protein PROCERA Acts in Guard Cells to Promote Stomatal Closure. The Plant Cell, 29(12), 3186-3197. doi:10.1105/tpc.17.00542Ohad, N., Shichrur, K., & Yalovsky, S. (2007). The Analysis of Protein-Protein Interactions in Plants by Bimolecular Fluorescence Complementation. Plant Physiology, 145(4), 1090-1099. doi:10.1104/pp.107.107284Ori, N., Cohen, A. R., Etzioni, A., Brand, A., Yanai, O., Shleizer, S., … Eshed, Y. (2007). Regulation of LANCEOLATE by miR319 is required for compound-leaf development in tomato. Nature Genetics, 39(6), 787-791. doi:10.1038/ng2036Pal, S., Zhao, J., Khan, A., Yadav, N. S., Batushansky, A., Barak, S., … Rachmilevitch, S. (2016). Paclobutrazol induces tolerance in tomato to deficit irrigation through diversified effects on plant morphology, physiology and metabolism. Scientific Reports, 6(1). doi:10.1038/srep39321Palatnik, J. F., Wollmann, H., Schommer, C., Schwab, R., Boisbouvier, J., Rodriguez, R., … Weigel, D. (2007). Sequence and Expression Differences Underlie Functional Specialization of Arabidopsis MicroRNAs miR159 and miR319. Developmental Cell, 13(1), 115-125. doi:10.1016/j.devcel.2007.04.012Parapunova, V., Busscher, M., Busscher-Lange, J., Lammers, M., Karlova, R., Bovy, A. G., … de Maagd, R. A. (2014). Identification, cloning and characterization of the tomato TCP transcription factor family. BMC Plant Biology, 14(1), 157. doi:10.1186/1471-2229-14-157Park, S. J., Jiang, K., Schatz, M. C., & Lippman, Z. B. (2011). Rate of meristem maturation determines inflorescence architecture in tomato. Proceedings of the National Academy of Sciences, 109(2), 639-644. doi:10.1073/pnas.1114963109Pharis, R. P., & King, R. W. (1985). Gibberellins and Reproductive Development in Seed Plants. Annual Review of Plant Physiology, 36(1), 517-568. doi:10.1146/annurev.pp.36.060185.002505Porri, A., Torti, S., Romera-Branchat, M., & Coupland, G. (2012). Spatially distinct regulatory roles for gibberellins in the promotion of flowering of Arabidopsis under long photoperiods. Development, 139(12), 2198-2209. doi:10.1242/dev.077164Preston, J. C., & Hileman, L. C. (2013). Functional Evolution in the Plant SQUAMOSA-PROMOTER BINDING PROTEIN-LIKE (SPL) Gene Family. Frontiers in Plant Science, 4. doi:10.3389/fpls.2013.00080Reinecke, D. M., Wickramarathna, A. D., Ozga, J. A., Kurepin, L. V., Jin, A. L., Good, A. G., & Pharis, R. P. (2013). Gibberellin 3-oxidase Gene Expression Patterns Influence Gibberellin Biosynthesis, Growth, and Development in Pea. PLANT PHYSIOLOGY, 163(2), 929-945. doi:10.1104/pp.113.225987Rubio-Somoza, I., & Weigel, D. (2011). MicroRNA networks and developmental plasticity in plants. Trends in Plant Science, 16(5), 258-264. doi:10.1016/j.tplants.2011.03.001Rubio-Somoza, I., Zhou, C.-M., Confraria, A., Martinho, C., von Born, P., Baena-Gonzalez, E., … Weigel, D. (2014). Temporal Control of Leaf Complexity by miRNA-Regulated Licensing of Protein Complexes. Current Biology, 24(22), 2714-2719. doi:10.1016/j.cub.2014.09.058Salinas, M., Xing, S., Höhmann, S., Berndtgen, R., & Huijser, P. (2011). Genomic organization, phylogenetic comparison and differential expression of the SBP-box family of transcription factors in tomato. Planta, 235(6), 1171-1184. doi:10.1007/s00425-011-1565-ySarvepalli, K., & Nath, U. (2011). Hyper-activation of the TCP4 transcription factor in Arabidopsis thaliana accelerates multiple aspects of plant maturation. The Plant Journal, 67(4), 595-607. doi:10.1111/j.1365-313x.2011.04616.xSerrano-Mislata, A., Bencivenga, S., Bush, M., Schiessl, K., Boden, S., & Sablowski, R. (2017). DELLA genes restrict inflorescence meristem function independently of plant height. Nature Plants, 3(9), 749-754. doi:10.1038/s41477-017-0003-yShikata, M., & Ezura, H. (2016). Micro-Tom Tomato as an Alternative Plant Model System: Mutant Collection and Efficient Transformation. Methods in Molecular Biology, 47-55. doi:10.1007/978-1-4939-3115-6_5Stettler, R. F. (1964). DOSAGE EFFECTS OF THE LANCEOLATE GENE IN TOMATO. American Journal of Botany, 51(3), 253-264. doi:10.1002/j.1537-2197.1964.tb06628.xVarkonyi-Gasic, E., Wu, R., Wood, M., Walton, E. F., & Hellens, R. P. (2007). Protocol: a highly sensitive RT-PCR method for detection and quantification of microRNAs. Plant Methods, 3(1), 12. doi:10.1186/1746-4811-3-12Vendemiatti, E., Zsögön, A., Silva, G. F. F. e, de Jesus, F. A., Cutri, L., Figueiredo, C. R. F., … Peres, L. E. P. (2017). Loss of type-IV glandular trichomes is a heterochronic trait in tomato and can be reverted by promoting juvenility. Plant Science, 259, 35-47. doi:10.1016/j.plantsci.2017.03.006Vicente, M. H., Zsögön, A., de Sá, A. F. L., Ribeiro, R. V., & Peres, L. E. P. (2015). Semi-determinate growth habit adjusts the vegetative-to-reproductive balance and increases productivity and water-use efficiency in tomato ( Solanum lycopersicum ). Journal of Plant Physiology, 177, 11-19. doi:10.1016/j.jplph.2015.01.003Wang, J.-W., Czech, B., & Weigel, D. (2009). miR156-Regulated SPL Transcription Factors Define an Endogenous Flowering Pathway in Arabidopsis thaliana. Cell, 138(4), 738-749. doi:10.1016/j.cell.2009.06.014Wilkie, J. D., Sedgley, M., & Olesen, T. (2008). Regulation of floral initiation in horticultural trees. Journal of Experimental Botany, 59(12), 3215-3228. doi:10.1093/jxb/ern188Yamaguchi, A., Wu, M.-F., Yang, L., Wu, G., Poethig, R. S., & Wagner, D. (2009). The MicroRNA-Regulated SBP-Box Transcription Factor SPL3 Is a Direct Upstream Activator of LEAFY, FRUITFULL, and APETALA1. Developmental Cell, 17(2), 268-278. doi:10.1016/j.devcel.2009.06.007Yamaguchi, N., Winter, C. M., Wu, M.-F., Kanno, Y., Yamaguchi, A., Seo, M., & Wagner, D. (2014). Gibberellin Acts Positively Then Negatively to Control Onset of Flower Formation in Arabidopsis. Science, 344(6184), 638-641. doi:10.1126/science.1250498Yamaguchi, S. (2008). Gibberellin Metabolism and its Regulation. Annual Review of Plant Biology, 59(1), 225-251. doi:10.1146/annurev.arplant.59.032607.092804Yamamoto, A., Nakamura, T., Adu-Gyamfi, J. J., & Saigusa, M. (2002). RELATIONSHIP BETWEEN CHLOROPHYLL CONTENT IN LEAVES OF SORGHUM AND PIGEONPEA DETERMINED BY EXTRACTION METHOD AND BY CHLOROPHYLL METER (SPAD-502). Journal of Plant Nutrition, 25(10), 2295-2301. doi:10.1081/pln-120014076Yanai, O., Shani, E., Russ, D., & Ori, N. (2011). Gibberellin partly mediates LANCEOLATE activity in tomato. The Plant Journal, 68(4), 571-582. doi:10.1111/j.1365-313x.2011.04716.xYu, S., Galvão, V. C., Zhang, Y.-C., Horrer, D., Zhang, T.-Q., Hao, Y.-H., … Wang, J.-W. (2012). Gibberellin Regulates the Arabidopsis Floral Transition through miR156-Targeted SQUAMOSA PROMOTER BINDING–LIKE Transcription Factors. The Plant Cell, 24(8), 3320-3332. doi:10.1105/tpc.112.101014Yuste-Lisbona, F. J., Quinet, M., Fernández-Lozano, A., Pineda, B., Moreno, V., Angosto, T., & Lozano, R. (2016). Characterization of vegetative inflorescence (mc-vin) mutant provides new insight into the role of MACROCALYX in regulating inflorescence development of tomato. Scientific Reports, 6(1). doi:10.1038/srep18796Zhang, S., Zhang, D., Fan, S., Du, L., Shen, Y., Xing, L., … Han, M. (2016). Effect of exogenous GA 3 and its inhibitor paclobutrazol on floral formation, endogenous hormones, and flowering-associated genes in ‘Fuji’ apple ( Malus domestica Borkh.). Plant Physiology and Biochemistry, 107, 178-186. doi:10.1016/j.plaphy.2016.06.005Zhang, X., Zou, Z., Zhang, J., Zhang, Y., Han, Q., Hu, T., … Ye, Z. (2010). Over-expression of sly-miR156a in tomato results in multiple vegetative and reproductive trait alterations and partial phenocopy of the sft mutant. FEBS Letters, 585(2), 435-439. doi:10.1016/j.febslet.2010.12.03

Downregulation of miR-31 in Diabetic Nephropathy and its Relationship with Inflammation

Background/Aims: There is a lack of reliable biological markers for the early diagnosis of diabetic nephropathy (DN) during type 2 diabetes. In this pilot study we aim to assess whether miR-31 levels are modulated by the presence of DN and whether the expression of this miRNA is related to leukocyte-endothelial interactions and inflammation. Methods: Thirty-one T2D patients were enrolled in this pilot study; 18 with no diabetic complications and 13 with diabetic nephropathy. 24 non-diabetic subjects and 13 T2D patients with retinopathy (absent of other complications) were included to test the specificity of miR-31. Following anthropometric and biochemical evaluation, serum miR-31 levels were assessed by Real Time-PCR. Leukocyte-endothelial interactions were evaluated by a parallel flow chamber in vitro model. Serum TNFα, IL-6 and ICAM-1 levels were determined by XMAP-technology in a flow cytometry-based Luminex 200 instrument. Results: Serum miR-31 levels were similar between control and T2D subjects. However, T2D patients with DN displayed reduced levels of miR-31 with respect to patients without complications. This decrease in miR-31 was more pronounced in patients with macroalbuminuria than in those with microalbuminuria and was specific for DN, since patients with retinopathy displayed unaltered miR-31 levels. The presence of DN involved a lower leukocyte rolling velocity and an increased rolling flux and adhesion. miR-31 levels were positively correlated with leukocyte rolling velocity and negatively associated to leukocyte adhesion, TNFα, IL-6 and ICAM-1 levels. Conclusion: Serum miR-31 may be a biomarker for DN in T2D patients. The regulation of this miRNA seems to be related to the recruitment of leukocytes to vascular walls induced by pro-inflammatory and adhesion molecules