Filogenómica del complejo Pseudomonas aeruginosa

Trabajo fin de máster en Bioinformática y Biología ComputacionalEl complejo de P. aeruginosa es un conjunto de bacterias pertenecientes al orden Pseudomonadales, unas γ-proteobacterias gram negativas que se encuentran en distintos nichos ecológicos y que está compuesto por una gran variedad de especies. A lo largo de los años, las especies se han definido en base a su morfología y sus actividades metabólicas. Después mediante la secuenciación de relojes filogenéticos como el 16S y por hibridación DNA-DNA para comparaciones de especies o cepas dos a dos, pero estas técnicas se están sustituyendo por métodos de caracterización genotípica utilizando una serie de genes marcadores que se encuentran en todas las especies. En esto se basan los Análisis Multilocus de Secuencia (MSLA en inglés), aunque existen otros métodos basados en el estudio del genoma completo para obtener filogenias más precisas que las que se obtienen mediante MLSA, tales como la identidad media de nucleótidos (Average Nucleotide Identity, ANI), ANIb (ANI como una identidad media de todas las coincidencias de blastn) y ANIm (ANI con el software MUMmer). En este trabajo se estudiaron las relaciones filogenéticas de las especies pertenecientes al complejo de P.aeruginosa utilizando MLSA y ANIb. Al mismo tiempo, también se definió el genoma central del complejo, el genoma principal de cada grupo, el genoma específico de cada especie, el genoma específico de cada grupo y el pangenoma gracias al estudio de los ortólogos, así como las distintas características funcionales que definan a cada grupo de P. aeruginosa

Dynamic-Catenal Phytosociology for evaluating vegetation

[EN] The conservation of nature is a problem that has concerned the scientific community for many years. Plants and plant communities play a main role in evaluation and land management studies, owing to their importance as natural and cultural resources. Several studies from the perspective of flora and vegetation have been carried out in the last fifty years (some of them directly related to Phytosociology). According to that, the Dynamic-Catenal Phytosociology must be considered as a very useful tool to evaluate the conservation status of vegetation and to establish suitable models for land management. The fundamental phytosociological concepts to take into account in the evaluation processes of the conservation status of vegetation are reviewed in this study

Study of the anticancer properties of optically active titanocene oximato compounds

New water soluble and optically active cyclopentadienyl titanium derivatives [(η5-C5H5)2Ti{(1R,4S)-ĸON,(R)NH}Cl] (R = Bn (Benzyl) 1a’, 2-pic (2-picolylamine) 1b’) have been synthesized. The novel compounds along with those previously described [(η5-C5H5)2Ti{(1S,4R)-ĸON,(R)NH}Cl] (R = Bn 1a, 2-pic 1b) were evaluated by polarimetry, ultra-violet and circular dichroism spectroscopy. The structure of 1b was determined by single crystal X-ray crystallography and showed a unique terminal monohapto Tisingle bondO disposition of the oximato ligand. All enantiomers have been tested against several cancer cell lines in vitro: prostate PC-3 and DU-145, lung A-549, pancreas MiaPaca-2, colorectal HCT-116, leukemia Jurkat and cervical HeLa. In addition, 1a, 1b and 1b’ were tested against non-tumorigenic prostate RWPE-1 cell line. After 24 h of incubation, 1b and 1b’ were moderately active against Jurkat and A-549 cells. The anti-proliferative effect of titanium compounds on prostate PC-3, DU-145 and RWPE-1 cell lines was also assessed after 72 h of drug exposure. The cytotoxic profile of the enantiomers was similar, exception made for the PC-3 cells, with S,R-isomers exhibiting cytotoxicities 2 to 3 times higher than R,S-compounds. Under these conditions, derivative 1b showed calculated IC50 values better than those of Tacke's Titanocene-Y (bis-[(p-methoxybenzyl)cyclopentadienyl]titanium(IV) dichloride) on both the prostate PC-3 and DU-145 cells. 1a and 1b cytotoxic behaviour shows certain selectiveness, with activities 2–4 times lower on normal prostate RWPE-1 than on cancer PC-3 cells. Furthermore, 1b produces higher cytotoxicity on prostate PC-3, DU-145 and RWPE-1 cells than the additive dose of titanocene dichloride and pro-ligand b·HCl. Additionally, compound-DNA interactions have been investigated by equilibrium dialysis, Fluorescence Resonance Energy Transfer (FRET) melting assays and viscometric titrations, which suggest that these metal complexes and/or their hydrolysis products bind DNA either in the minor groove or externally.Ministerio de Economía y CompetitividadComunidad Autónoma de MadridUniversidad de Alcal

Applications of bioclimatology to assess effects of climate change on viticultural suitability in the DO León (Spain)

[EN] Spain accounts for 13.1% of the world’s vineyard area, and viticulture is crucial for the socioeconomic and cultural sectors. Vineyards are among the perennial crops that can suffer most negative impacts under climate change which can pose challenges to the sustainability of viticulture. Local and regional studies are needed to assess these impacts to help implement effective strategies in response to climate change. To this end, our approach involves integrating both conventional agroclimatic indices and those new bioclimatic indices that have proven to be essential for the characterization and demarcation of vineyards into species distribution models to assess areas suitable for viticulture under climate change projections. The proposed methodology was tested in a viticultural region located in northwestern Spain (DO León). An ensemble platform was used to build consensus models encompassing three general circulation models, two emission scenario pathways and two time horizons. Only the predictors that effectively characterize each grape variety were included in the models. The results revealed increases in the continentality index, compensated thermicity index, hydrothermic index of Branas, and temperature range during ripening in all the future scenarios analyzed in comparison to current conditions. Conversely, the values for the annual ombrothermic index and growing season precipitation may decrease in the future. The pattern of changes for 2070 will be more pronounced than for 2050. A significant loss of future habitat suitability was detected within the limits of the study area for the grape varieties analyzed. This negative impact could be counteracted to some degree with new and favorable areas for the cultivation of vineyards in territories located at the north of the DO limits. We suggest that our results could help policymakers to develop practices and strategies to conserve existing grape varieties and to implement efficient adaptation measures for mitigating or anticipating the effects of climate change on viticulture.SIThis work was supported by the Regional Ministry of Education, Junta de Castilla y León (Spain), EDU/667/201

Biological evaluation of water soluble arene Ru(II) enantiomers with amino-oxime ligands

New water soluble, enantiopure arene ruthenium compound SRuSN-(1R,4S)-[(η6-p-cymene)Ru{ĸNH(Bn),ĸNOH}Cl]Cl (Bn = benzyl, 1a′) has been synthesized. The novel compound along with that previously described RRuRN-(1S,4R)-[(η6-p-cymene)Ru{ĸNH(Bn),ĸNOH}Cl]Cl (1a) was evaluated by polarimetry, ultra-violet and circular dichroism spectroscopy. The structure of novel ruthenium derivative 1a′ was determined by single crystal X-ray crystallography. Both enantiomers have been tested against several cancer cell lines in vitro: prostate PC-3, lung A-549, pancreas MIA PaCa-2, colorectal HCT-116, leukemia Jurkat and cervical HeLa. Both enantiomers are active and versatile cytotoxic agents, showing IC50 values from 2 to 12 times lower than those found for cisplatin in the different cell lines evaluated. The mechanism of cell death induced by the metal compounds was analyzed in A-549 and Jurkat cell lines. Derivatives 1a and 1a′ induced apoptotic cell death of A-549 cells while dose-dependent cell death mechanisms have been found in the Jurkat cell line. Compound-DNA interactions have been investigated by equilibrium dialysis, Fluorescence Resonance Energy Transfer (FRET) melting assays and viscometric titrations, revealing moderate binding affinity of 1a and 1a′ towards duplex DNA. Finally, the efficacy of 1a in a preliminary in vivo assay of PC-3 xenografts in nude mice has been evaluated, resulting in a promising inhibition of tumor growth by 45%. Analysis of tumor tissue also showed a significant decrease of levels of crucial molecules in the invasive phenotype of PC-3 cells.Ministerio de Economía y CompetitividadComunidad Autónoma de MadridUniversidad de Alcal

Dynamic-Catenal Phytosociology for Evaluating Vegetation

The conservation of nature is a problem that has concerned the scientific community for many years. Plants and plant communities play a main role in evaluation and land management studies, owing to their importance as natural and cultural resources. Several studies from the perspective of flora and vegetation have been carried out in the last fifty years (some of them directly related to Phytosociology). According to that, the Dynamic-Catenal Phytosociology must be considered as a very useful tool to evaluate the conservation status of vegetation and to establish suitable models for land management. The fundamental phytosociological concepts to take into account in the evaluation processes of the conservation status of vegetation are reviewed in this study

BRAF activation by metabolic stress promotes glycolysis sensitizing NRASQ61-mutated melanomas to targeted therapy

Glycolysis; Melanomas; Targeted therapyGlucólisis; Melanomas; Terapia dirigidaGlucòlisi; Melanomes; Teràpia dirigidaNRAS-mutated melanoma lacks a specific line of treatment. Metabolic reprogramming is considered a novel target to control cancer; however, NRAS-oncogene contribution to this cancer hallmark is mostly unknown. Here, we show that NRASQ61-mutated melanomas specific metabolic settings mediate cell sensitivity to sorafenib upon metabolic stress. Mechanistically, these cells are dependent on glucose metabolism, in which glucose deprivation promotes a switch from CRAF to BRAF signaling. This scenario contributes to cell survival and sustains glucose metabolism through BRAF-mediated phosphorylation of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase-2/3 (PFKFB2/PFKFB3). In turn, this favors the allosteric activation of phosphofructokinase-1 (PFK1), generating a feedback loop that couples glycolytic flux and the RAS signaling pathway. An in vivo treatment of NRASQ61 mutant melanomas, including patient-derived xenografts, with 2-deoxy-D-glucose (2-DG) and sorafenib effectively inhibits tumor growth. Thus, we provide evidence for NRAS-oncogene contributions to metabolic rewiring and a proof-of-principle for the treatment of NRASQ61-mutated melanoma combining metabolic stress (glycolysis inhibitors) and previously approved drugs, such as sorafenib.This work was funded by Instituto de Salud Carlos III and co-funded by European Union (ERDF/ESF, “A way to make Europe”/“Investing in your future”) PI14/0375-Fondos FEDER J.A.R., PI17/00043-Fondos FEDER; J.A.R., PI20/0384-Fondos FEDER; J.A.R., Euronanomed2-ISCIII (AC16/00019)-Fondos FEDER; J.A.R., Asociación Española Contra el Cancer (AECC-GCB15152978SOEN) (supported P.G.M., K.M.); J.A.R., Ramón Areces Foundation (supported K.M. and research); J.A.R. (PI17/00412)-Fondos FEDER; R.B., A.M., A.N.S. We thank A. Zorzano’s laboratory for technical assistance and performance of Seahorse technology

Ruminal fermentation and biohydrogenation in ewes fed with sainfoin pellets under water restriction

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Estudios de asociación del genoma completo (GWAS) con biomarcadores sanguíneos y de lana para identificar genes de tolerancia al estrés hídrico y térmico en ovejas de raza Rasa aragonesa

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