The CBRB regulon: Promoter dissection reveals novel insights into the CbrAB expression network in Pseudomonas putida

CbrAB is a high ranked global regulatory system exclusive of the Pseudomonads that responds to carbon limiting conditions. It has become necessary to define the particular regulon of CbrB and discriminate it from the downstream cascades through other regulatory components. We have performed in vivo binding analysis of CbrB in P. putida and determined that it directly controls the expression of at least 61 genes; 20% involved in regulatory functions, including the previously identified CrcZ and CrcY small regulatory RNAs. The remaining are porines or transporters (20%), metabolic enzymes (16%), activities related to protein translation (5%) and orfs of uncharacterised function (38%). Amongst the later, we have selected the operon PP2810-13 to make an exhaustive analysis of the CbrB binding sequences, together with those of crcZ and crcY. We describe the implication of three independent non-palindromic subsites with a variable spacing in three different targets; CrcZ, CrcY and operon PP2810-13 in the CbrAB activation. CbrB is a quite peculiar σN—depen-dent activator since it is barely dependent on phosphorylation for transcriptional activation. With the depiction of the precise contacts of CbrB with the DNA, the analysis of the multi-merisation status and its dependence on other factors such as RpoN o IHF, we propose a model of transcriptional activation.Ministerio de Economía y Competitividad BIO2014-57545-

Síntesis, caracterización y aplicación de zeolitas CuHZSM-5 con porosidad jerarquizada para su uso como trampa de hidrocarburos durante el arranque en frío de motores

El presente proyecto fin de carrera tiene como objetivo final la creación de zeolitas jerárquicas para aplicarlas en la reducción de la contaminación ambiental producida por los hidrocarburos emitidos por los coches, en particular el propeno y el tolueno. Para conseguirlo se probaron zeolitas ZSM-5 como trampas de hidrocarburos. Estas zeolitas habían sido jerarquizadas previamente y luego intercambiadas con Cu(NO3)2•3H2O. La jerarquización de la porosidad consiste en la creación de mesoporos en la estructura de la zeolita inicialmente microporosa mediante la realización de un tratamiento alcalino, en este caso utilizando como base NaOH. Con este tratamiento se consigue disolver el silicio y extraerlo así de la estructura dando lugar a poros de mayor tamaño, los mesoporos. Para optimizar esa parte del proceso se fijó la concentración de la base y se variaron la duración y la temperatura (parámetros de mayor influencia). El ratio Si/Al también es fundamental cuando se realiza un tratamiento alcalino ya que numerosos estudios demuestran que la presencia del Al en la estructura inhibe la extracción del Si, con lo cual si se reduce el número de aluminios por silicio se consigue una mayor formación de mesoporos. Para estar seguros del éxito en la creación de mesoporosidad se realizaron experimentos con distintos ratios (Si/Al 25 y Si/Al 40) cuyos resultados podíamos comparar para sacar conclusiones. Por otro lado, se quería optimizar el intercambio con Cu(NO3)2•3H2O para obtener mejores resultados durante las pruebas en el reactor, ya que trabajos previos del equipo habían demostrado que el cobre incluido en la zeolita mejoraba las prestaciones de las trampas de hidrocarburos (HC). Para ello se probaron intercambios con distintas concentraciones de Cu(NO3)2•3H2O para dos zeolitas ZSM-5 distintas, una con ratio Si/Al 25, la otra con Si/Al 40. El ratio Si/Al es fundamental en el intercambio de cationes porque limita la cantidad de éstos que se puede introducir en los espacios que se forman entre la estructura. Una vez optimizado el proceso de Desilicación y establecida la concentración ideal para el intercambio con cobre, se inició la síntesis de las muestras de la investigación principal con las distintas condiciones para el tratamiento alcalino, dejando la zeolita en su forma NaZSM-5. Después se realizó un ataque ácido para eliminar el aluminio extraído de la estructura durante el tratamiento alcalino y así recuperar el ratio Si/Al disminuido tras la disolución del silicio. Para devolver la zeolita a su forma ácida es necesario realizar un intercambio con NH4NO3 y calcinar para obtener la forma HZSM5 que permite realizar el intercambio para incluir el cobre. Con los tratamientos terminados se probaron las zeolitas en un reactor de lecho fijo que simulaba las condiciones de un motor durante el arranque enfrío, este test se denomina CST (Cold Start Test). Por último, se caracterizaron, cristalinidad, morfología, porosidad y acidez y se realizaron análisis de los resultados obtenidos en los CST. Con todos los análisis se concluye que las condiciones óptimas de producción de la zeolita CuHZSM-5 mediante desilicación e intercambio iónico para su uso en gases de motores durante el arranque en frío son: un tratamiento alcalino con NaOH 0,2 M, a 65 °C durante 15 min, seguido de un ataque ácido con HCl 0,1 M y un intercambio iónico con Cu(NO3)2•3H2O 100 mM. La zeolita producida en estas condiciones no sólo mejora la capacidad de adsorción de la zeolita ZSM-5 microporosa sino también la actividad catalítica y por lo tanto, la capacidad de tratamiento de propeno y tolueno en su aplicación como trampa de HC

Undercarboxylated Osteocalcin: A Promising Target for Early Diagnosis of Cardiovascular and Glycemic Disorders in Patients with Metabolic Syndrome: A Pilot Study

Lifestyle changes are causing an exponential increase in the prevalence of obesity and metabolic syndrome (MetS) worldwide. The most frequent complications of these are the development of diabetes (T2D) and cardiovascular disease (CVD). Accurate tools are needed to classify the cardiovascular risk (CVR) in the MetS population. In recent years, numerous biomarkers of bone metabolism have been associated with CVR. The aim of this study was to determine the levels of undercarboxylated osteocalcin (ucOC) in a cohort of patients with MetS and to analyse its association with MetS parameters and CVR as well as with T2D prevalence. A longitudinal study was conducted in which a MetS population was followed for one year. Weight change, adherence to the Mediterranean diet (MedDiet), ucOC levels, MetS parameters and CVR were analysed and CVR was calculated using different scores. Our results showed a decrease of CVR associated with a better adherence to the MetDiet resulting in higher HDL-C and ucOC levels though the improvement of MetS risk factors. This bone protein appeared as a potential biomarker to classify CVR in the MetS population, especially for MetS patients without prevalent T2D. Furthermore, ucOC serum levels could be good predictors of T2D prevalence.Instituto de Salud Carlos III European Commission PI18-00803 PI21/01069 PI18-01235 FI19/00118 CD20/00022European CommissionJunta de Andalucia CD20/00022 PI-0268-2019 RH-0069202

((R)-( )-3-Hydroxyquinuclidium)[FeCl4]; a plastic hybrid compound with chirality, ferroelectricity and long range magnetic ordering

Quinuclidinium salts and their derivatives are now in the focus of materials science as building units of multifunctional materials. Their properties can be easily switchable, allowing their use in a wide range of physical applications. One type of these kinds of materials, the homochiral hybrid halometallate ferroelectric compounds, is not well understood. In this work, (R)-( )-3-quinuclidinol hydrochloride was used in the synthesis of ((R)-( )-3-hydroxyquinuclidium)[FeCl4]. The use of this enantiomeric cation forces crystallographic non-centrosymmetry, which was confirmed by polarimetry and circular dichroism spectroscopy. We studied the physical properties of this compound at different temperatures by single crystal, synchrotron and neutron powder X-ray diffraction, which showed a rich series of structural and magnetic phase transitions. From synchrotron powder X-ray diffraction data, a plastic phase was observed above 370 K (phase I). Between 370 K and ca. 310 K, an intermediate polar phase was detected, solved in a non-centrosymmetric polar space group (C2) (phase II). Below ca. 310 K, the compound crystallizes in the triclinic P1 non-centrosymmetric space group (phase III) which is maintained down to 4 K, followed by phase IV, which shows tridimensional magnetic ordering. The temperature evolution of the neutron diffraction data shows the appearance of new reflections below 4 K. These reflections can be indexed to a commensurate propagation vector k = (0, 0, 12). The magnetic structure below TN was solved in the Ps1 Shubnikov space group, which gives rise to an antiferromagnetic structure, compatible with the magnetometry measurements. Near room temperature, the crystal phase transition is associated with a dielectric change. In particular, the phase transition between phase III (S.G.:P1) and phase II (S.G.:C2) involves an increase of symmetry between two non-centrosymmetric space groups. Therefore, it allows, by symmetry, the emergence of ferroelectric and ferroelastic ordering. Piezoresponse force microscopy (PFM) imaging measurements provided evidence for polarization switching and a local ferroelectric behavior of phase III at room temperature. Additionally, the obtained butterfly curve and hysteresis loop by PFM exhibits a low coercive voltage of B10 V. This value is remarkable, since it approaches those obtained for materials with application in ferroelectric random access memories (FeRAMs).Financial support from Universidad de Cantabria (Proyecto Puente convocatoria 2018 funded by SODERCAN_FEDER), Universidad del País Vasco/Euskal Herriko Unibertsitatea (GIU17/50 and PPG17/37) and Ministerio de Economia y Competividad (MAT2017-89239-C2-(1,2)-P, MAT2017-83631-C3-3-R, MAT2017-86453-R, PGC2018-097520-A-100 and PID2019-104050RAI00) is acknowledged. The authors gratefully acknowledge the technical and human support provided by SGIKer (UPV/EHU, MINECO, GV/EJ, ERDF, and ESF). Carmen Martín is grateful to VI PPIT-2018 from Universidad de Sevilla. The paper is (partly) based on the results of experiments carried out at the ALBA Synchrotron Light Source in Barcelona (proposal 2019083666) and Institute Laue-Langevin (ILL) of Grenoble (Proposals 5-31-2580 and 5-31-2460)

Investigación joven con perspectiva de género VI

Actas del VI Congreso Internacional de Jóvenes Investigadorxs con perspectiva de género (Getafe, 16 - 18 de junio de 2021) organizado por el Instituto Universitario de Estudios de Género de la Universidad Carlos III de Madrid.El concepto de la vejes en la literatura griega arcaica / María Secades Fonseca se inscribe en el marco del Proyectos de I+D+i del Ministerio de Ciencia e Innovación, titulado Vulnerabilidad intrafamiliar y política en el mundo antiguo y dirigido por Susana Reboreda Morillo y Rosa María Cid López (Ref. PID2020-116349GB-I00).Generando una interpretación del Derecho en clave de igualdad de género / Alicia Cárdenas Cordón y Gloria Serrano Valverde es un trabajo elaborado y presentado en el marco del Proyecto de Investigación RTI2018-10669-B-100 ‘‘GEN-DER: Generando una interpretación del Derecho en clave de igualdad de género’’, enmarcado en el Programa Estatal de I+D+i Orientada a los Retos de la Sociedad- Ministerio de Ciencia e Innovación (2019-2021)

Investigación joven con perspectiva de género III

Actas del III Congreso Internacional de Estudios de Género, Perspectivas y Retos de Futuro: Jóvenes Investigadores (Getafe, 13 - 15 de junio de 2018) organizado por el Instituto Universitario de Estudios de Género de la Universidad Carlos III de Madrid

Normothermic regional perfusion vs. super-rapid recovery in controlled donation after circulatory death liver transplantation

[Background & Aims] Although there is increasing interest in its use, definitive evidence demonstrating a benefit for postmortem normothermic regional perfusion (NRP) in controlled donation after circulatory death (cDCD) liver transplantation is lacking. The aim of this study was to compare results of cDCD liver transplants performed with postmortem NRP vs. super-rapid recovery (SRR), the current standard for cDCD.[Methods] This was an observational cohort study including all cDCD liver transplants performed in Spain between June 2012 and December 2016, with follow-up ending in December 2017. Each donor hospital determined whether organ recovery was performed using NRP or SRR. The propensity scores technique based on the inverse probability of treatment weighting (IPTW) was used to balance covariates across study groups; logistic and Cox regression models were used for binary and time-to-event outcomes.[Results] During the study period, there were 95 cDCD liver transplants performed with postmortem NRP and 117 with SRR. The median donor age was 56 years (interquartile range 45–65 years). After IPTW analysis, baseline covariates were balanced, with all absolute standardised differences <0.15. IPTW-adjusted risks were significantly improved among NRP livers for overall biliary complications (odds ratio 0.14; 95% CI 0.06–0.35, p <0.001), ischaemic type biliary lesions (odds ratio 0.11; 95% CI 0.02–0.57; p = 0.008), and graft loss (hazard ratio 0.39; 95% CI 0.20–0.78; p = 0.008).[Conclusions] The use of postmortem NRP in cDCD liver transplantation appears to reduce postoperative biliary complications, ischaemic type biliary lesions and graft loss, and allows for the transplantation of livers even from cDCD donors of advanced age.[Lay summary] This is a propensity-matched nationwide observational cohort study performed using livers recovered from donors undergoing cardiac arrest provoked by the intentional withdrawal of life support (controlled donation after circulatory death, cDCD). Approximately half of the livers were recovered after a period of postmortem in situ normothermic regional perfusion, which restored warm oxygenated blood to the abdominal organs, whereas the remainder were recovered after rapid preservation with a cold solution. The study results suggest that the use of postmortem normothermic regional perfusion helps reduce rates of post-transplant biliary complications and graft loss and allows for the successful transplantation of livers from older cDCD donors.Peer reviewe

Identificación del efecto de la dependencia de densidad, las prácticas agrícolas y las variables climáticas en la dinámica poblacional de las malas hierbas

Comunicación presentada en el XIV Congreso Nacional de Malgerbología, celebrado en Valencia del 5 al 7 de noviembre de 2013.La teoría de la dinámica de poblaciones define factores endógenos y exógenos que influyen en los cambios temporales en la densidad de las plantas. Sin embargo, engeneral, se desconoce la contribución relativa a cada uno de ellos. El objetivo de este trabajo fue evaluar el efecto de la dependencia de la densidad, las p´racticas agrícolas y el clima en la dinámica a largo plazo de 7 malas hierbas. Se ha encontrado que la dependencia de la densidad reguló a 4 y 6 y, la temperatura únicamente a 1 y 2 de las 7 especies bajo mínomo y no laboreo, respectivamente, destacando la importancia de los factores endógenos como reguladores poblacionales.Peer Reviewe

Novel insights into the CbrAB expression network in Pseudomonas putida

Resumen del trabajo presentado en el 8th Congress of European Microbiologists (FEMS), celebrado en Glasgow (Escocia), del 7 al 11 de julio de 2019Background: The ability to recognize and convert external environmental stimuli into appropriate physiological responses is of fundamental importance for all organisms. In Pseudomonads, the two-component regulatory system CbrAB is responsible to the adaptation to carbon limitation. Global analysis of the regulatory element CbrB in P. putida shows that it controls a collection of ¿N-dependent genes involved in the assimilation of several amino acids but also in a number of other physiological functions, such as chemotaxis or stress tolerance. Conversely, very little is known about the signal that triggers the Cbr response, which is channeled through the histidine kinase CbrA. CbrA represents a new family of sensor HKs as its structure suggests it may link signalling to transport of a molecule. Its N terminus contains a 13 TM domain with similarity to the sodium-proline symporter PutP from E. coli, which is connected to the C terminal catalytic HK domain which could regulate transport through the sodium/solute symporter domain. Objectives: Describe the CbrB regulon for a consensus CbrB-binding sequence description and determine its contribution of in the transcriptional activation. Also , explore the regulation of CbrA expression and the implication of the TM, PAS and HK domains in the signal transduction. Methods: ChIP-seq analysis and point mutagenesis to the DNA sequences necessary for its binding and transcriptional activation. Also, we explore the control of cbrA expression and constructed truncated versions of CbrA to follow the signal transduction. Results: Altogether, we show an extensive view on the molecular mechanism of CbrAB activation