A Quantum Dot-Based FLIM Glucose Nanosensor

In the last few years, quantum dot (QD) nanoparticles have been employed for bioimaging and sensing due to their excellent optical features. Most studies have used photoluminescence (PL) intensity-based techniques, which have some drawbacks, especially when working with nanoparticles in intracellular media, such as fluctuations in the excitation power, fluorophore concentration dependence, or interference from cell autofluorescence. Some of those limitations can be overcome with the use of time-resolved spectroscopy and fluorescence lifetime imaging microscopy (FLIM) techniques. In this work, CdSe/ZnS QDs with long decay times were modified with aminophenylboronic acid (APBA) to achieve QD-APBA conjugates, which can act as glucose nanosensors. The attachment of the boronic acid moiety on the surface of the nanoparticle quenched the PL average lifetime of the QDs. When glucose bonded to the boronic acid, the PL was recovered and its lifetime was enhanced. The nanosensors were satisfactorily applied to the detection of glucose into MDA-MB-231 cells with FLIM. The long PL lifetimes of the QD nanoparticles made them easily discernible from cell autofluorescence, thereby improving selectivity in their sensing applications. Since the intracellular levels of glucose are related to the metabolic status of cancer cells, the proposed nanosensors could potentially be used in cancer diagnosis.This research was funded by grants CTQ2014-56370-R and CTQ2017-85658-R from the Spanish Ministerio de Ciencia, Innovación y Universidades, Agencia Estatal de Investigación (AEI), and the European Regional Development Fund (ERDF)

Protein O-Fucosyltransferase 1 Undergoes Interdomain Flexibility in Solution

Protein O-fucosyltransferase 1 (PoFUT1) is a GT-B fold enzyme that fucosylates proteins containing EGF-like repeats. GT-B glycosyltransferases have shown a remarkable grade of plasticity adopting closed and open conformations as a way of tuning their catalytic cycle, a feature that has not been observed for PoFUT1. Here, we analyzed Caenorhabditis elegans PoFUT1 (CePoFUT1) conformational behavior in solution by atomic force microscopy (AFM) and single-molecule fluorescence resonance energy transfer (SMF-FRET). Our results show that this enzyme is very flexible and adopts mainly compact conformations and to a lesser extend a highly dynamic population that oscillates between compact and highly extended conformations. Overall, our experiments illustrate the inherent complexity of CePoFUT1 dynamics, which might play a role during its catalytic cycle.ARAID: MEC (CTQ2013-44367-C2-2-P, BFU2016-75633-P and PID2019-105451GBI00 to RH-G, CTQ2017-85658-R and CTQ2014-56370-R to AO)Gobierno de Aragón (E35_R20 and LMP58_18)FEDER (2014-2020) funds for ‘Building Europe from Aragón’Juan de la Cierva fellowship IJCI-2017-3287

Circularly Polarized Luminescence of [6]Helicenes through Excited- State Intramolecular Proton Transfer

This project received funding from the European Research Council (ERC) under the European Union's Horizon 2020 research and innovation programme (Grant Agreements No. 716139 and No. 677023), the Swiss National Science Foundation (SNSF, PP00P2_170534, PP00P2_198900), Project PGC2018-101181-B-I00 funded by MCIN/AEI/ 10.13039/501100011033 FEDER "Una manera de hacer Europa", grant A-FQM-230-UGR20 funded by FEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades and the German Academic Exchange Service (DAAD, postdoc fellowship to D. G.). Open access funding provided by the University of Zurich.We present the concept of combining circularly polarized luminescence (CPL) and excited-state intramolecular proton transfer (ESIPT) features into a single molecule as a strategy to generate high-performance ESIPT-based CPL materials. For this purpose, a [6]helicene bearing two ESIPT structural units was synthesized using a double Suzuki-Miyaura reaction and a double C(sp(2))-H hydroxylation approach. The photophysical properties of the doubly hydroxylated [6]helicene were studied in parallel with a non-hydroxylated [6]helicene control compound, revealing that the presence of a chiral [6]helicene unit results in a strong CPL response and the presence of the ESIPT units in a considerable red shift. The red-shifted emission along with the outstanding g(lum) (approximate to 10(-2)) and a large Stokes shift makes the doubly hydroxylated [6]helicene a promising candidate for use in optoelectronics.European Research Council (ERC) 716139 677023Swiss National Science Foundation (SNSF) PP00P2_170534 PP00P2_198900Deutscher Akademischer Austausch Dienst (DAAD)University of ZurichMCIN/AEI FEDER "Una manera de hacer Europa" PGC2018-101181-B-I00FEDER/Junta de Andalucia-Consejeria de Transformacion Economica, Industria, Conocimiento y Universidades A-FQM-230-UGR2

Mitochondrial pH Nanosensors for Metabolic Profiling of Breast Cancer Cell Lines

We thank the company Nanogetic S.L. (Granada, Spain) for support with the synthesis of the SS peptides and the Centro de Instrumentacion Cientifica (CIC) of the Universidad de Granada for use of the TEM facilities.The main role of mitochondria, as pivotal organelles for cellular metabolism, is the production of energy (ATP) through an oxidative phosphorylation system. During this process, the electron transport chain creates a proton gradient that drives the synthesis of ATP. One of the main features of tumoral cells is their altered metabolism, providing alternative routes to enhance proliferation and survival. Hence, it is of utmost importance to understand the relationship between mitochondrial pH, tumoral metabolism, and cancer. In this manuscript, we develop a highly specific nanosensor to accurately measure the intramitochondrial pH using fluorescence lifetime imaging microscopy (FLIM). Importantly, we have applied this nanosensor to establish differences that may be hallmarks of different metabolic pathways in breast cancer cell models, leading to the characterization of different metabophenotypes.Spanish Ministerio de Ciencia, Innovacion y Universidades CTQ2014-56370-R CTQ2017-86568-REuropean Union (EU)Spanish Agencia Estatal de InvestigacionFundacion Ramon Arece

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

Novel ortho-OPE metallofoldamers: binding-induced folding promoted by nucleating Ag(i)-alkyne interactions

We have developed a new family of ortho-oligophenylene ethynylene (o-OPE) metallofoldamers. The folding of these helicates is induced by nucleating carbon-metal interactions between Ag(i) cations and the alkynes of the inner core of the o-OPEs. These o-OPEs form metal-organic assemblies where at least three alkyne moieties are held in close proximity to form novel Ag(i)-complexes with the metal ion lodged into the helical cavity. NMR titration experiments and photokinetic studies have provided quantitative data about the thermodynamic and kinetic features of such binding/folding phenomena. X-ray diffraction and DFT studies have been performed to extract structural information on how the Ag(i) cation is accommodated into the cavity. The great simplicity and versatility of these new metallofoldamers open up the possibility to develop novel structures with applications in material science and/or in asymmetric catalysisThis research was funded by the Regional Government of Andalucía (project P09-FQM-4571) and the ICIQ Foundation. DM thanks Regional Government of Andalucía for her contract. AML thanks MICINN for her FPU fellowship. The authors thank the Centro de Servicios de Informática y Redes de Comunicaciones (CSIRC), Universidad de Granada, for providing the computing tim

Optosensores en flujo : de la regeneración a la renovación de la superficie sensora

[EN] In this Memory it is showed the development of ten spectroscopic continuous flow sensors for the automatic determination of organic and inorganic species of environmental and pharmaceutical interest. These sensors have been classified according to the developed methodology in sensors with or without renovation of the sensing surface. Three flow sensors without renovation of the sensing surface ha ve been developed using this methodology, one of them uses spectrophotometric detection and two of them spectrofluorimetric detection, based on the measurement of the absorbance in UV region or the intrinsic fluorescence of the analytes. In all these cases it is not necessary previous derivation reactions. The seven developed sensors with renovation of the sensing surface are based on the concept of Bead Injection Spectroscopy. The use of this methodology is justified when the species of interest is strongly retained on the sensing surface and the regeneration of the support is not possible in suitable conditions or becomes extraordinarily difficult. The methodology used in these seven sensors is based on the injection of an exact volume of a homogenous beads suspension in a flow injection analysis system with spectroscopic detection. The beads (solid support) are loaded in the flow cell placed in the detection zone of a nondestructive spectroscopic detector. At the end of the analysis, the beads are discarded by reversing the flow and instantaneously transported out of the system, being replaced the sensing surface after each measurement. In all cases, a great study of the experimental variables that can affect this type of sensors has been performed. Next, the sensors were calibrated for the corresponding analytes, finding a calibration equation that allowed relating the values of the analytical signals with the concentration of the sample. The method used in all the cases was the univariante ca libration. Studies of interferences were made to know the response of the sensor in presence of the organic and inorganic compounds that usually accompany the analytes in the rea l samples. In the case of serious interferences occurring, appropriate modifications for their elimination were designed. Finally, the proposed sensors were applied to the determination of these analytes in samples of different nature. The results confirm the validity of the proposed sensors.Tesis Univ. Jaén. Departamento de Química Física y Analítica. Leída el 21 de enero de 200

Uso de Tecnologías de la Información y la Comunicación (TIC) para el aprendizaje multisensorial de personas con discapacidad o necesidades educativas especiales

Se ha preparado un material docente, haciendo uso de metodologías TIC, que facilita la accesibilidad a la información para el conjunto del alumnado con discapacidad visual, y/o necesidades educativas especiales, en concreto para el temario de la asignatura Química General, del grado en Nutrición Humana y Dietética, impartido en la Facultad de Farmacia. Los temarios digitalizados, disponibles para el alumnado a través de la plataforma SWAD, se acompañarán por el mismo material en formato de audio, lo que origina una metodología docente multisensorial. Los archivos de audio están disponibles en el repositorio institucional de la Universidad de Granada.Unidad de Innovación Docente (Universidad de Granada). Proyecto número 10-31

Breast Cancer Cell Subtypes Display Different Metabolic Phenotypes That Correlate with Their Clinical Classification

Recent studies on cancer cell metabolism have achieved notable breakthroughs that have led to a new scientific paradigm. How cancer cell metabolic reprogramming is orchestrated and the decisive role of this reprogramming in the oncogenic process and tumor adaptative evolution has been characterized at the molecular level. Despite this knowledge, it is essential to understand how cancer cells can metabolically respond as a living whole to ensure their survival and adaptation potential. In this work, we investigated whether different cancers and different subtypes display different metabolic phenotypes with a focus on breast cancer cell models representative of each clinical subtype. The potential results might have significant translational implications for diagnostic, prognostic and therapeutic applications.Metabolic reprogramming of cancer cells represents an orchestrated network of evolving molecular and functional adaptations during oncogenic progression. In particular, how metabolic reprogramming is orchestrated in breast cancer and its decisive role in the oncogenic process and tumor evolving adaptations are well consolidated at the molecular level. Nevertheless, potential correlations between functional metabolic features and breast cancer clinical classification still represent issues that have not been fully studied to date. Accordingly, we aimed to investigate whether breast cancer cell models representative of each clinical subtype might display different metabolic phenotypes that correlate with current clinical classifications. In the present work, functional metabolic profiling was performed for breast cancer cell models representative of each clinical subtype based on the combination of enzyme inhibitors for key metabolic pathways, and isotope-labeled tracing dynamic analysis. The results indicated the main metabolic phenotypes, so-called ‘metabophenotypes’, in terms of their dependency on glycolytic metabolism or their reliance on mitochondrial oxidative metabolism. The results showed that breast cancer cell subtypes display different metabophenotypes. Importantly, these metabophenotypes are clearly correlated with the current clinical classifications.Spanish Agencia Estatal de Investigación (Ministry of Science and Innovation)European Regional Development Fund [grant numbers CTQ2014-56370-R and CTQ2017-85658-R]Fundación Ramón ArecesInitiative "Solidaridad Entre Montañas

Dataset from article ‘ Breast cancer cell subtypes display different metabolic phenotypes that correlate with their clinical classification’

Metabolic reprogramming of cancer cells represents an orchestrated network of evolving molecular and functional adaptations during oncogenic progression. In particular, how metabolic reprogramming is orchestrated in breast cancer and its decisive role in the oncogenic process and tumor evolving adaptations are well consolidated at the molecular level. Nevertheless, potential correlations between functional metabolic features and breast cancer clinical classification still represent issues that have not been fully studied to date. Accordingly, we aimed to investigate whether breast cancer cell models representative of each clinical subtype might display different metabolic phenotypes that correlate with current clinical classifications. In the present work, functional metabolic profiling was performed for breast cancer cell models representative of each clinical subtype based on the combination of enzyme inhibitors for key metabolic pathways, and isotope-labeled tracing dynamic analysis. The results indicated the main metabolic phenotypes, so-called ‘metabophenotypes’, in terms of their dependency on glycolytic metabolism or their reliance on mitochondrial oxidative metabolism. The results showed that breast cancer cell subtypes display different metabophenotypes. Importantly, these metabophenotypes are clearly correlated with the current clinical classifications.Funding: This research, including APC charges, was funded by the Spanish Agencia Estatal de Investigación (Ministry of Science and Innovation) and the European Regional Development Fund [grant numbers CTQ2014-56370-R and CTQ2017-85658-R]; the Fundación Ramón Areces; and the initiative Solidaridad Entre Montañas