14 research outputs found
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
Efficient acetate sensor in biological media based on a selective Excited State Proton Transfer (ESPT) reaction
We have synthesized a new fluoride−containing xanthenic dye able to dynamically and
quantitatively detect acetate anion, a biologically relevant analyte, in water. We studied
deeply the photophysical properties of the compound and verified its use as an acetate
probe in synthetic serum.MINECO-FEDER CTQ2014-53598Junta
Andalucía FQM2012-79
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
Photosensitizing properties and subcellular localisation of 3,4‑dihydro‑β‑carbolines harmaline and harmalol
Harmaline (1) and harmalol (2) represent two 3,4-dihydro-β-carboline (DHβCs) most frequently reported in a vast number
of living systems. Fundamental aspects including the photosensitizing properties, cellular uptake, as well as the cyto- and
phototoxicity of 1 and 2 were investigated herein. The molecular basis underlying the investigated processes are elucidated.
Data reveal that both alkaloids show a distinctive pattern of extracellular DNA photodamage. Compound 1 induces a DNA
photodamage profile dominated by oxidised purines and sites of base loss (AP sites), whereas 2 mostly induces single-strand
breaks (SSBs) in addition to a small extent of purine oxidative damage. In both cases, DNA oxidative damage would occur
through type I mechanism. In addition, a concerted hydrolytic attack is suggested as an extra mechanism accounting for the
SSBs formation photoinduced by 2. Subcellular internalisation, cyto- and phototoxicity of 1 and 2 and the corresponding
full-aromatic derivatives harmine (3) and harmol (4) also showed quite distinctive patterns in a structure-dependent manner.
These results are discussed in the framework of the potential biological, biomedical and/or pharmacological roles reported
for these alkaloids
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