Self-Assembled Lanthanide Antenna Glutathione Sensor for the Study of Immune Cells

This work was supported by grants CTQ2017-85658-R, BFU2015-67284-R, and PID2019-104366RB-C22 funded by MCIN/AEI/10.13039/501100011033/FEDER "Una manera de hacer Europa"; grant PID2020-114256RB-I00 funded by MCIN/AEI/10.13039/501100011033; grant A-FQM-386-UGR20 funded by FEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades, and the CSIC grant 201580E073. Funding for open access charge: Universidad de Granada/CBUA.The small molecule 8-methoxy-2-oxo-1,2,4,5- tetrahydrocyclopenta[de]quinoline-3-carboxylic acid (2b) behaves as a reactive non-fluorescent Michael acceptor, which after reaction with thiols becomes fluorescent, and an efficient Eu3+ antenna, after self-assembling with this cation in water. This behavior makes 2b a highly selective GSH biosensor, which has demonstrated high potential for studies in murine and human cells of the immune system (CD4+ T, CD8+ T, and B cells) using flow cytometry. GSH can be monitored by the fluorescence of the product of addition to 2b (445 nm) or by the luminescence of Eu3+ (592 nm). 2b was able to capture baseline differences in GSH intracellular levels among murine and human CD4+ T, CD8+ T, and B cells. We also successfully used 2b to monitor intracellular changes in GSH associated with the metabolic variations governing the induction of CD4+ naiv̈ e T cells into regulatory T cells (TREG).MCIN/AEI/FEDER "Una manera de hacer Europa" CTQ2017-85658-R BFU2015-67284-R PID2019-104366RB-C22MCIN/AEI PID2020-114256RB-I00FEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades A-FQM-386-UGR20CSIC grant 201580E073Universidad de Granada/CBU

A solvatofluorochromic silicon-substituted xanthene dye useful in bioimaging

In this work, we have performed an in-depth study of the photophysics and solvatofluorochromism of a red-emitting Si-xanthene dye, an analog of Tokyo Magenta (TM) historically developed by Egawa et al. (Chem. Commun. 2011, 47, 4162–4164). The results show a strong dependency of the emission properties of 2-Me-4-OMe-TM on the polarity of the solvent. For instance, the dye exhibited an increase in its fluorescence lifetime with the decrease in solvent polarity. Therefore, in this work, this spectral behavior has been used as a new approach for determining the intracellular microenvironment polarity through the measurement of its fluorescence lifetime by Fluorescence Lifetime Imaging Microscopy (FLIM). Our experiments confirmed the ability of the dye to detect changes in polarity between different intracellular compartments.This work was funded by grants CTQ2017-85658-R, CTQ2014-55474-C2-2-R (Spanish Ministry of Economy and Competitiveness; Agencia Estatal de Investigacion, AEI; and European Regional Development Fund, ERDF), QM2012-790 (Junta de Andalucía), and a grant from the Fundación Botín

Application of new information and communication methodologies (ICT) for laboratory teaching in applied physics and physical chemistry

En este artículo se presenta un trabajo de innovación en metodología docente que tiene como objetivo la mejora en el aprendizaje de las enseñanzas prácticas de laboratorio de la asignatura de Física Aplicada y Fisicoquímica de la Licenciatura de Farmacia de la Universidad de Granada. La enseñanza práctica de diversas asignaturas de esta facultad tradicionalmente se ha realizado con la colaboración de los “monitores de prácticas”, los cuales se encargan de ayudar a sus compañeros en la realización de las prácticas de laboratorio, estando siempre coordinados y supervisados por un profesor. Puesto que este sistema tiene sus carencias y limitaciones, con la metodología presentada en este trabajo se pretende que desaparezca la figura del monitor de prácticas persiguiendo al mismo tiempo que el alumno sea más receptivo y muestre mayor interés. La experiencia nos demuestra que resulta fundamental la visualización de la manipulación y las operaciones necesarias para la realización de una práctica. Las nuevas tecnologías audiovisuales de la información y la comunicación (TIC) ponen al servicio de la enseñanza herramientas muy didácticas que hacen más atractivo el aprendizaje. Con tal fin, se ha elaborado para la asignatura antes mencionada un vídeo de cada práctica en formato DVD, en los que, con objeto de guiar al alumno en el trabajo experimental, se muestra la manipulación del material y reactivos empleados en cada práctica, uso de la instrumentación específica, buenas prácticas en un laboratorio y adquisición y análisis de datos. El objetivo que se persigue es que el alumno realice la práctica simultáneamente al desarrollo del vídeo, con la opción de interactuar con el vídeo y con el propio profesor presente en la sesión. En definitiva, mediante esta metodología de prácticas innovadora se pretende la captación de la atención del alumno, su motivación, promoción del trabajo autónomo y capacidad de iniciativa facilitando de esta forma su capacidad de autoaprendizaje. Todo lo comentado, contribuye a potenciar la adaptación de los actuales estudios universitarios al Espacio Europeo de Educación Superior.The aim of this innovative teaching work is the improvement in the learning process of laboratory practical sessions of the subject ‘Applied Physics and Physical Chemistry’ of the Pharmacy degree syllabuses at the University of Granada. The laboratory practical sessions of several subjects in this Faculty have been traditionally carried out with the collaboration of ‘laboratory demonstrators’, who help to their classmates in the performance of the experiments, being always coordinated and project, we expect on one hand the figure of ‘laboratory demonstrators’ to disappear, and on the other hand to achieve that the student is more receptive and shows a higher interest in the practical sessions. Our experience proves that it is very important for the student to visualize the manipulation and methodology needed for the performance of the experiments in the laboratory. The new audiovisual Information and Communication Technologies (ICT) provide very didactic tools, which make the learning more attractive. With this purpose, we have prepared a DVD movie of each experiment planned in the practical sessions of the subject. These movies show the manipulation of the material and reagents, the use of specific instrumentation, good behaviour in the laboratory, and data acquisition and analysis, with the aim of guiding the student in the experimental work. The student will perform the experiments simultaneously to the movie projection, with the possibility of interacting with the movie itself and the senior professor present during the session. In short, we expect to enhance the motivation and own-initiative of the students, facilitating their self-learning, by means of an innovative teaching methodology that contributes to promote the adaptation to the European Space for Higher Education.El proyecto ha sido financiado por la Unidad de Innovación Docente de la Universidad de Granada (Proyecto de Innovación Docente, Código Nº 08-109

N-Methyl-β-carboline alkaloids: structuredependent photosensitizing properties and localization in subcellular domains

N-Methyl-β-carboline (βC) alkaloids, including normelinonine F (1b) and melinonine F (2b), have been found in a vast range of living species playing different biological, biomedical and/or pharmacological roles. Despite this, molecular bases of the mechanisms through which these alkaloids would exert their effect still remain unknown. Fundamental aspects including the photosensitizing properties and intracellular internalization of a selected group of N-methyl-βC alkaloids were investigated herein. Data reveal that methylation of the βC main ring enhances its photosensitizing properties either by increasing its binding affinity with DNA as a biomolecular target and/or by increasing its oxidation potential, in a structure- dependent manner. As a general rule, N(9)-substituted βCs showed the highest photosensitizing efficiency. With the exception of 2-methyl-harminium, all the N-methyl-βCs investigated herein induce a similar DNA photodamage profile, dominated largely by oxidized purines. This fact represents a distinctive behavior when comparing with N-unsubstituted-βCs. On the other hand, although all the investigated compounds might accumulate mainly into the mitochondria of HeLa cells, methylation provides a distinctive dynamic pattern for mitochondrial uptake. While rapid (passive) diffusion is most probably reponsible for the prompt uptake/release of neutral βCs, an active transport appears to mediate the (reatively slow) uptake of the quaternary cationic βCs. This might be a consequence of a distinctive subcellular localization (mitochondrial membrane and/or matrix) or interaction with intracellular components. Biomedical and biotechnological implications are also discussed herein.ANPCyT PICT-2015-0374 2016-0370 2018-3193Spanish Ministry of Economy and Competitiveness CTQ201785658-R CTQ2014-55474-C2-2-

Self-Assembled Lanthanide Antenna Glutathione Sensor for the Study of Immune Cells

The small molecule 8-methoxy-2-oxo-1,2,4,5- tetrahydrocyclopenta[de]quinoline-3-carboxylic acid (2b) behaves as a reactive non-fluorescent Michael acceptor, which after reaction with thiols becomes fluorescent, and an efficient Eu3+ antenna, after self-assembling with this cation in water. This behavior makes 2b a highly selective GSH biosensor, which has demonstrated high potential for studies in murine and human cells of the immune system (CD4+ T, CD8 + T, and B cells) using flow cytometry. GSH can be monitored by the fluorescence of the product of addition to 2b (445 nm) or by the luminescence of Eu3+ (592 nm). 2b was able to capture baseline differences in GSH intracellular levels among murine and human CD4 + T, CD8 + T, and B cells. We also successfully used 2b to monitor intracellular changes in GSH associated with the metabolic variations governing the induction of CD4+ naï ve T cells into regulatory T cells (TREG ).This work was supported by grants CTQ2017-85658-R, BFU2015-67284-R, and PID2019-104366RB-C22 funded by MCIN/AEI/10.13039/501100011033/FEDER “Una manera de hacer Europa”; grant PID2020-114256RB-I00 funded by MCIN/AEI/10.13039/501100011033; grant A-FQM-386- UGR20 funded by FEDER/Junta de Andalucía-Consejería de Transformación Económica, Industria, Conocimiento y Universidades, and the CSIC grant 201580E073. Funding for open access charge: Universidad de Granada/CBUA.Peer reviewe

Desarrollo de una metodología innovadora asistida por tecnologías de la información y la comunicación (TIC) para el aprendizaje de prácticas de química física

Código Nº 08-109Los vídeos usan música con Licencia Creative Commons. Los álbunes utilizados son "blue for a soldier" y "hello pretty woman", ambos de Persson.Elaboración de vídeos en tiempo real con demostraciones de cómo realizar las prácticas: - manipulación de reactivos y material básico de laboratorio - instrumentación específica - buen comportamiento - tratamiento de residuos - tratamiento de datos para cálculos y resultados Objetivos generales: favorecer el autoaprendizaje e introducir en la docencia una metodología de prácticas innovadoras, potenciando la adaptación al Espacio Europeo de Educación Superior.Universidad de Granada. Departamento de Química-Física. Unidad de Innovación Docente

Prodecimiento para la estimación de la concentración de fosfatos en células vivas, colorante xantético y síntesis del mismo

Número de publicación: ES2474916 B1. Número de solicitud: 201330861.La invención se refiere a un método para estimar la concentración de los fosfatos presentes en las células vivas por medio de la medición el tiempo de decaimiento de la fluorescencia de un colorante de xanteno añadió a las células y se sometieron a una excitación por medio de luz láser pulsado. La invención también se refiere a un nuevo colorante xanteno derivado de fluoresceína, que tiene mejores propiedades espectrales que los ya conocidos, y a un método para la síntesis de dicho colorante.Universidad de Granad

Versatile synthesis and enlargement of functionalized distorted heptagon-containing nanographenes

Highly distorted polycyclic aromatic hydrocarbons (PAHs) are predicted to be attractive goals in nanoscience owing to the new properties they can exhibit. We have shown that a variety of functionalized distorted heptagon-containing nanographenes can be easily prepared from simple building blocks by a sequence of Co-catalyzed cyclotrimerization and cyclodehydrogenation reactions. The versatility of this strategy allows easy subsequent enlargement of these nanostructures by Ni-catalyzed cross-coupling and final cyclodehydrogenation reactions. Soluble extended distorted nanographenes 1 and 2 containing heptagon and an edge-shared pentagon-heptagon combination have been synthesized. High distortion of the polycyclic backbone of 2 caused by non-hexagonal rings and a helicene moiety was confirmed by X-ray crystallography. Experimental data reveal promising optical and electronic properties for distorted PAHs with long fluorescence lifetimes (up to 14.5 ns) and low band gaps (down to 2.27 eV). This straightforward and versatile synthetic strategy, the observed long fluorescence lifetimes and the small optical and electrochemical band gaps for the presented compounds may promote the future implementation of distorted graphene molecules in electronic devices.This project has received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation program (ERC-2015-STG-677023 - NANOGRAPHOUT). We thank the Ministerio de Economía y Competitividad (MINECO, Spain) (CTQ2014-53598-R, TEC2014-59730). A. G. C. and B. B. gratefully acknowledge funding from MINECO for RyC-2013-12943 and RYC-2012-10381 contracts. I. R. M. thanks MECD (Spain) for a predoctoral FPU fellowship. D. C. L. thanks Intramural CSIC project (201530E01). We also thank IZO-SGI SGIker of UPV/EHU for their technical and human support and computational resources provided by the RES and the CSIRC-Alhambra supercomputing facilities