Examinando por Autor "Bravo, S."
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Ítem Contrasting responses of cluster roots formation induced by phosphorus and nitrogen supply in Embothrium coccineum populations from different geographical origin(Springer Science and Business Media Deutschland GmbH, 2020-08) Bertin-Benavides, A.; Bascuñán-Godoy, L.; Henríquez-Castillo C.; Delgado, M.; Burgos, C.F.; Mardones, C.; Ávila-Valdés, A.; Valdebenito, F.; Bravo, S.; Rubilar, R.; Hasbún, R.; Zúñiga-Feest, A.Aims: Embothrium coccineum is a pioneer tree that produces cluster roots (CR) induced by phosphorus (P) or nitrogen (N) deficiency, but the role which both N and P play in CR formation for different E. coccineum populations is still unknown. We hypothesized that in E. coccineum, N influences the CR formation response, primarily induced by P limitation. However, this response depends on local adaptations to their specific conditions of geographic origin. Methods: Seedlings from three contrasting edaphoclimatic conditions (Northern, Central and Southern populations) were grown in hydroponic culture under high and low N supply and at five different P supply. Morphophysiological responses, including CR number, growth, biomass, P and N plant concentration, were studied. Results: Our results showed that at high N supply, CR formation was stimulated with increased P supply, but just until it reaches a maximum of 5 μM, above this concentration CR formation decreases. However, at low N supply, a constitutive response was observed regardless of P supply. Moreover, each of these responses depends on the populations. Conclusions: Our study suggests that N drives CR formation induced by P supply and that a differential response exists among E. coccineum populations, that could be related to their origin edaphoclimatic conditions. © 2020, Springer Nature Switzerland AG.Ítem Enhanced CRAd activity using enhancer motifs driven by a nucleosome positioning sequence(Cell Press, 2013) Bravo, S.; Núñez, F.; Cruzat, F.; Cafferata, E.; De Ferrari, G.; Montecino, M.; Podhajcer, O.Cancer development involves changes driven by the epigenetic machinery, including nucleosome positioning. Recently, the concept that adenoviral replication may be driven by tumor specific promoters (TSPs) gained support, and several conditionally replicative adenoviruses (CRAd) exhibited therapeutic efficacy in clinical trials. Here, we show for the first time that placing a nucleosome positioning sequence (NPS) upstream of a TSP combined with Wnt-responsive motifs (pART enhancer) enhanced the TSP transcriptional activity and increased the lytic activity of a CRAd. pART enhanced the transcriptional activity of the gastrointestinal cancer (GIC)-specific REG1A promoter (REG1A-pr); moreover, pART also increased the in vitro lytic activity of a CRAd whose replication was driven by REG1A-Pr. The pART enhancer effect in vitro and in vivo was strictly dependent on the presence of the NPS. Indeed, deletion of the NPS was strongly deleterious for the in vivo antitumor efficacy of the CRAd on orthotopically established pancreatic xenografts. pART also enhanced the specific activity of other heterologous promoters; moreover, the NPS was also able to enhance the responsiveness of hypoxia- and NFκB-response elements. We conclude that NPS could be useful for gene therapy approaches in cancer as well as other diseases.Ítem Reversible in vivo cellular changes occur during desiccation and recovery: Desiccation tolerance of the resurrection filmy fern Hymenophyllum dentatum Cav.(Facultad de Ciencias Naturales y Oceanográficas, Universidad de Concepción, 2016-12) Bravo, S.; Parra, M.J.; Castillo, R.; Sepúlveda, F.; Turner, A.; Bertín, A.; Osorio, G.; Tereszczuk, J.; Bruna, C.; Hasbún, R.The present work explores in vivo physiological, morphological and chemical features during full hydration, desiccation and rehydration of the filmy fern Hymenophyllum dentatum with two main objectives: 1) to get further insight about the mechanisms underlying its desiccation tolerance, and 2) to understand how this plant manages mechanical stress induced by water loss and recovery. With these purposes, physiological (relative water content and Fv/Fm chlorophyll fluorescence parameter), morphological (Confocal Laser Scanning Microscopy and 3D reconstruction) and chemical (FTIR microspectroscopy) data were obtained and compared between fully hydrated, desiccated and rehydrated tissues of H. dentatum. Remarkable changes in cell architecture and chemical composition were observed in vivo in desiccated leaves. Cells were smaller, showed a collapsed general appearance, and were delimitated by apparently folded cell walls. Marked changes in chloroplasts location and decrease in the number of active chloroplasts were also evidenced. Chemical experiments showed that changes in the secondary structure of proteins and in the polysaccharide composition of the cell wall occur in desiccated cells. All changes were rapidly reversed upon rehydration. This study shows that H. dentatum presents an extreme case of desiccation tolerance, able to withdraw severe, rapid and consecutive dehydration/rehydration induced stress by the function of constitutive systems of protection and reparation, in which cell wall folding plays a relevant role as a protective system against mechanical and oxidative stress. Besides, H. dentatum is proposed as an excellent plant model for the study of dissection tolerance such as one cell layer fern, auto-fluorescence of cellular compartments, and simple long term storage under laboratory conditions, among others.Ítem Wnt/β-Catenin Signaling Enhances Cyclooxygenase-2 (COX2) Transcriptional Activity in Gastric Cancer Cells(Public Library of Science, 2011) Nuñez, F.; Bravo, S.; Cruzat, F.; Montecino, M.; Ferrari, G.Increased expression of the cyclooxygenase-2 enzyme (COX2) is one of the main characteristics of gastric cancer (GC), which is a leading cause of death in the world, particularly in Asia and South America. Although the Wnt/β-catenin signaling pathway has been involved in the transcriptional activation of the COX2 gene, the precise mechanism modulating this response is still unknown. Methodology/Principal Findings: Here we studied the transcriptional regulation of the COX2 gene in GC cell lines and assessed whether this phenomenon is modulated by Wnt/β-catenin signaling. We first examined the expression of COX2 mRNA in GC cells and found that there is a differential expression pattern consistent with high levels of nuclear-localized β-catenin. Pharmacological treatment with either lithium or valproic acid and molecular induction with purified canonical Wnt3a significantly enhanced COX2 mRNA expression in a dose- and time-dependent manner. Serial deletion of a 1.6 Kbp COX2 promoter fragment and gain- or loss-of-function experiments allowed us to identify a minimal Wnt/β-catenin responsive region consisting of 0.8 Kbp of the COX2 promoter (pCOX2-0.8), which showed maximal response in gene-reporter assays. The activity of this pCOX2-0.8 promoter region was further confirmed by site-directed mutagenesis and DNA-protein binding assays. Conclusions/Significance: We conclude that the pCOX2-0.8 minimal promoter contains a novel functional T-cell factor/lymphoid enhancer factor (TCF/LEF)-response element (TBE Site II; -689/-684) that responds directly to enhanced Wnt/β-catenin signaling and which may be important for the onset/progression of GC.