Aprendizaje colaborativo y el modelo de clase invertida como ayuda para el aprendizaje del metabolismo

Comunicación oral y poster en el Grupo de la Enseñanza de la Bioquímica de la Sociedad Española de Bioquímica y Bioqlogía MolecularLo siguiente resume el contenido de la presentación oral que será expuesta en este Congreso dedicada a describir la base de las metodologías de aprendizaje colaborativo (AC) y de inversión de la clase (flipped learning) en la mejora de la enseñanza/aprendizaje de la Bioquímica. Preguntados a los alumnos muchos consideran esta materia difícil por ser demasiado amplia, no tener tiempo para estudiarla e integrar sus contenidos, y donde los profesores no dedicamos suficiente tiempo a hacer ejercicios en clase. La transición de un modelo educativo centrado en la enseñanza hacia el que emana del EEES que pone su foco en el aprendizaje significativo y la adquisición de competencias, supone una importante renovación metodológica. Dicha renovación pasa por complementar las tradicionales clases magistrales, donde el alumno adopta un papel pasivo, por metodologías activas que estimulen su participación, la colaboración entre pares y el trabajo autónomo. El AC tiene su base en el constructivismo social, que entiende el aprendizaje como un proceso social que se construye no sólo con el profesor, sino también con los compañeros, el contexto y el significado que se asigna a lo que se aprende. Además de favorecer más que la enseñanza tradicional el rendimiento académico, el AC permite adquirir importantes competencias transversales. No obstante, la puesta en marcha del AC exige al profesor nuevas competencias y más esfuerzo que impartir clases magistrales. Por ello, y por la idea de que se verán obligados a renunciar a partes del temario muchos profesores no se plantean estrategias de AC en la docencia de la Bioquímica. En cualquier caso existen buenas alternativas metodológicas para “ganar tiempo” sin sacrificar temario. Una de ellas, es el aprendizaje inverso, con la que se consigue además satisfacer una de las exigencias de los estudiantes, el hacer más ejercicios en clase.[This work was supported by PIE17-145 project (UMA). The attendance to the Workshop on Biochemical Education within the 41 SEBBM Congress (September 2018, Santander) has received a grant from "I Plan Propio Integral de Docencia (UMA)”

Learning contract, co-operative and flipped learning as useful tools for studying metabolism

Es el Abstract de una comunicación a un congreso internacional sobre educaciónUndergraduate students in Biology identify Metabolic Biochemistry as a particularly difficult subject. This is due to the fact that students need to interconnect properly all the contents of its syllabus throughout their study of the subject in order to get a global insight of the complex regulatory features controlling metabolic pathways within the metabolic network under different physiologic and pathologic conditions, as well as metabolism as a whole. Due to these objective difficulties, a high percentage of our students face the study of this subject as a very hard task beyond their forces and capacities. This perception leads to high rates of premature dropout. In previous years, less than 40% of all the registered students attended the examinations of Metabolic Biochemistry (a subject in the second year of the Degree of Biology at our University). Even worse, less than 25% of our students passed the exams. From the academic year 2015/16 on, we are developing innovative teaching projects (PIE15-163 and PIE17-145, funded by University of Malaga) aimed to increase our student loyalty to the subject (and hence to increase their attendance to exams) and to help them to learn more effectively metabolism and its regulation. These innovative teaching projects are based on the use of several powerful tools: a learning contract and problem-based learning within the framework of group tasks promoting an actual collaborative learning in a flipped classroom. The present communication will show the implementation of the PIE15-163 and PIE17-145 projects and some results obtained from them.This work was supported by Malaga University funds granted to the educational innovation project PIE17-145. The attendance to the END2018 International Conference on Education and New Developments (June 2018, Budapest, Hungary) has received a grant from "I Plan Propio Integral de Docencia. Universidad de Málaga"]

Changes in the nuclear proteomic profiling of human glioblastoma cells after glutaminase overexpression

A proteomics study of the nuclear protein profiling of human glioblastoma cells with targeted glutaminase expression. The study was performed with protein microarrays and SELDI-TOF technology.Human glutaminase (GA) enzymes are the products of two genes, GLS and GLS2. GA isozymes play markedly different roles in tumour biology: GLS isoforms (KGA and GAC) are related to cell growth and proliferation, whereas GLS2 isoforms (GAB and LGA) are associated with low proliferation rates and are characteristics for resting or quiescent cells. Furthermore, the GAB isoform has been recently involved in transcriptional regulation. Due to the proposed roles of GAB isoform in cellular differentiation and transcriptional modulation, the aim of the present study is to profile differentially expressed nuclear proteins in order to discover putative biomarkers associated with GAB overexpression in glioma cells. Nuclear cell proteins were incubated with strong anion exchange (Q10) and weak cation exchange (CM10) ProteinChip arrays and analyzed using surface-enhanced laser desorption/ionization time-of- flight mass spectrometry (SELDI-TOF/MS) proteomics technology. T98G-GAB nuclear protein expression profiles were compared with those of T98G-WT and empty vector transfected T98G-pcDNA3 with the ProteinChip Data Manager Client 4.0 software. Eighteen proteins were found to be differentially expressed between control cells and GAB-transfected T98G cells. Nine proteins with m/z between 5.5-29.7 were identified to be highly expressed in T98G-GAB (P≤ 0.01), while the expression of two proteins with m/z values of 7.8 and 8.5 were higher in control cells (P≤ 0.01). Our study shows the potential of proteomics profiling to get a deep insight into the role of nuclear GAB in brain tumors in order to assess its suitability as a novel anti-cancer therapeutic target. References Pérez-Gómez C, et al. Co-expression of glutaminase K and L isoenzymes in human tumour cells. Biochem. J. 2005; 386, 535-42. Szeliga M, et al. Transfection with liver-type glutaminase (LGA) cDNA alteres gene expression and reduces viability, migration and proliferation of T98G glioma cells. Glia 2009; 57, 1014-23.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech.This work was supported by Grant SAF2010-17573 from the Ministry of Science and Innovation of Spain, Excellence Grant CVI-6656 from the regional Andalusian government (Junta de Andalucia), and Grant RD06/1012 of the RTA RETICS network from the Spanish Health Institute Carlos III

Potential protective role of reactive astrocytes in the periventricular parenchyma in congenital hydrocephalus

Background Cerebrospinal fluid accumulation in hydrocephalus produces an elevation of intraventricular pressure with pathological consequences on the periventricular brain parenchyma including ischemia, oedema, oxidative stress, and accumulation of metabolic waste products. Here we studied in the hyh mouse, an animal model of congenital hydrocephalus, the role of reactive astrocytes in this clinical degenerative condition. Materials and Methods Wild type and hydrocephalic hyh mice at 30 days of postnatal age were used. Three metabolites related to the oxidative and neurotoxic conditions were analysed in ex vivo samples (glutathione, glutamine and taurine) using High Resolution Magic Angle Spinning (HR-MAS). Glutathione synthetase and peroxidase, glutamine synthetase, kidney-type glutaminase (KGA), and taurine/taurine transporter were immunolocated in brain sections. Results Levels of the metabolites were remarkably higher in hydrocephalic conditions. Glutathione peroxidase and synthetase were both detected in the periventricular reactive astrocytes and neurons. Taurine was mostly found free in the periventricular parenchyma and in the reactive astrocytes, and the taurine transporter was mainly present in the neurons located in such regions. Glutamine synthetase was found in reactive astrocytes. Glutaminase was also detected in the reactive astrocytes and in periventricular neurons. These results suggest a possible protective response of reactive astrocytes against oxidative stress and neurotoxic conditions. Conclusions Astrocyte reaction seems to trigger an anti-oxidative and anti-neurotoxic response in order to ameliorate pathological damage in periventricular areas of the hydrocephalic mice.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. PI15-00619 to AJJ

A problem-/case-based learning approach as an useful tool for studying glycogen metabolism and its regulation

Versión preprint del manuscrito de los autores, publicado finalmente en: Biochemistry and Molecular Biology Education, con DOI: 10.1002/bmb.21449Metabolism and its regulation is one of the most complex and difficult topics for students learning biochemistry. A problem-/case-based learning (PBL) approach can be useful to help biochemistry students to fulfill the goal of acquiring an integrated view of metabolism and its regulation. The present article describes our experience enrolling volunteer students to learn glycogen metabolism making use of a design-based research methodology to develop teaching learning sequences focused on a PBL approach. Enrolled undergraduate students had better final scores than those students that did not participates. Furthermore, enrolled students were satisfied with the experience, finding it interesting, formative, and challenging.This work was supported by the University of Málaga (Spain) with funds granted to the educational innovation projects PIE15-163, PIE17-145, and PIE19-057. The experimental work carried out by our group is supported by grants PID2019-105010RB-I00 and EDU2017-82197-P (Spanish Ministry of Science, Innovation and Universities), UMA18-FEDERJA-220 (Andalusian Government and FEDER) and funds from group BIO 267 (Andalusian Government), as well as funds from “Plan Propio de Investigación y Transferencia” (U. Málaga)

Ratones knock-out del receptor lpa1 de ácido lisofosfatídico presentan un acusado déficit de la isoenzima glutaminasa KGA (GLS) y una morfología alterada en las espinas dendríticas de hipocampo y corteza

Objectives: The objective of the present study was to utilize mice with knocked-down lysophosphatidic acid 1 (LPA1) receptor to ascertain changes in glutamatergic transmission that may help to explain part of the cognitive and memory deficits shown by these KO-LPA1 mice. Material & methods: A well characterized KO-LPA1 mouse strain was used as animal model and compared with wild-type (WT) and heterozygous animals. Expression studies were implemented by immunohistochemistry and Western analysis of mouse brain regions, real-time quantitative RT-PCR of GA isoforms, enzymatic analysis of regional GA activity and Golgi staining to assess dendritic spine morphology and density. Results: A strong reduction of KGA immunoreactivity was mostly revealed in cerebral cortex and hippocampus of KO-LPA1 mice versus WT and heterozygous animals. In contrast, neither mRNA levels nor enzyme activity were significantly altered in KO mice suggesting compensatory mechanisms for neurotransmitter Glu synthesis. Interestingly, Golgi staining of hippocampal and cortical neurons revealed a clear morphology change toward a less-mature undifferentiated spine phenotype, without changes in the total number of spines. Conclusions: The molecular mechanisms underlying KGA downregulation in null LPA1 mutant mice are unknown. However, LPA increases neuronal differentiation, arborization and neurite outgrowth of developing neurons, while Gln-derived Glu, through GA reaction, has been also involved in neuronal growth and differentiation. It is tempting to speculate that downregulation of KGA protein in KO-LPA1 mice induce morphological changes in dendritic spines of cortical and hippocampal neurons which, in turn, may account for memory and cognitive deficits shown by KO-LPA1 mice.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Acknowledgements: Red de Trastornos Adictivos, RTA, (RD12/0028/0013/) RETICS, ISCIII, y Consejería Innovación, Ciencia y Empresa, Junta de Andalucía (Proyecto de Excelencia CVI-6656)

La enseñanza del metabolismo: retos y oportunidades

En el marco del Proyecto de Innovación Educativa de la Universidad de Málaga PIE15-163, cuya descripción y resultados incluimos, decidimos que esta era una excelente oportunidad para reflexionar acerca de la enseñanza del metabolismo y de poner por escrito dichas reflexiones en un libro. Quisimos y pudimos contar con la colaboración de buena parte de los compañeros del Departamento de Biología Molecular y Bioquímica que apoyaron con su firma el proyecto PIE15-163 y extendimos nuestra invitaciones a otros compañeros de dentro y fuera de la Universidad de Málaga. Del Departamento de Biología Molecular y Bioquímica de la Universidad de Málaga hemos recibido aportaciones de los catedráticos Victoriano Valpuesta Fernández, Ana Rodríguez Quesada y Antonio Heredia Bayona, los profesores titulares María Josefa Pérez Rodríguez, José Luis Urdiales Ruiz e Ignacio Fajardo Paredes y la investigadora postdoctoral y profesora sustituta interina Beatriz Martínez Poveda. De otros departamentos de la Universidad de Málaga hemos contado con las aportaciones de la catedrática del Departamento de Especialidades Quirúrgicas, Bioquímica e Inmunología Pilar Morata Losa, del catedrático del Departamento de Lenguajes y Ciencias de la Computación José Francisco Aldana Montes y los componentes de su grupo de investigación Khaos Ismael Navas Delgado, María Jesús García Godoy, Esteban López Camacho y Maciej Rybinski, del catedrático Ángel Blanco López, del Área de Conocimiento de Didáctica de las Ciencias Experimentales y del Doctor en Ciencias Químicas y actual doctorando del Programa de Doctorado "Educación y Comunicación Social" Ángel Luis García Ponce. De fuera de la Universidad de Málaga, hemos contado con las aportaciones del catedrático de la Universidad de La Laguna Néstor V. Torres Darias, de la catedrática de la Universitat de les Illes Balears Pilar Roca Salom y de sus compañeros los profesores Jorge Sastre Serra y Jordi Oliver, de los catedráticos de la Universidad de Granada Rafael Salto González y María Dolores Girón González y su colaborador el Dr. José Dámaso Vílchez Rienda, del profesor titular de la Universidad de Alcalá Ángel Herráez, del investigador postdoctoral de la Universidad de Erlangen (Alemania) Guido Santos y del investigador postdoctoral de la empresa Brain Dynamics Carlos Rodríguez Caso.Hemos estructurado los contenidos del libro en diversas secciones. La primera presenta el Proyecto en cuyo marco se ha gestado la iniciativa que ha conducido a la edición del presente libro. La segunda sección la hemos titulado "¿Qué metabolismo?" e incluye diversas aportaciones personales que reflexionan acerca de qué metabolismo debe conocer un graduado en Bioquímica, en Biología, en Química, en Farmacia o en Medicina, así como una aportación acerca de qué bioquímica estructural y enzimología son útiles y necesarias para un estudiante que vaya a afrontar el estudio del metabolismo. La tercera sección, "Bases conceptuales", analiza las aportaciones del aprendizaje colaborativo, el contrato de aprendizaje y el aprendizaje basado en la resolución de casos prácticos a la mejora del proceso enseñanza-aprendizaje dentro del campo de la Bioquímica y Biología Molecular, más concretamente en el estudio del metabolismo. La cuarta sección se titula "Herramientas", es la más extensa e incluye las diversas aportaciones centradas en propuestas concretas de aplicación relevantes y útiles para la mejora de la docencia-aprendizaje del metabolismo. Sigue una sección dedicada a presentar de forma resumida los "Resultados" del proyecto PIE15-163. El libro concluye con una "coda final" en la que se reflexiona acerca del aprendizaje de la Química a la luz de la investigación didáctica.Patrocinado por el Proyecto de Innovación Educativa de la Universidad de Málaga PIE15-16

Turning around Cycles: An Approach Based on Selected Problems/Cases to Stimulate Collaborative Learning about Krebs and His Four Metabolic Cycles

Metabolism is a challenging subject for bioscience students due to the intrinsic complexity of the metabolic network, as well as that of the overlapping mechanisms of metabolic regulation. Collaborative learning based on a problem-based learning approach can help students to successfully learn and understand metabolism. In the present article, we propose a selection of exercises, problems, and cases aimed to focus students’ attention on the scientific work made by Sir Hans Krebs and his collaborators to elucidate four main metabolic cycles, as well as on the study of these cycles, their regulation, and their metabolic integration. The objectives, the tools, and the implementation of this proposal are described, and the results obtained during its first implementation with volunteer students enrolled in two courses on metabolic regulation at our university are presented and discussed. These volunteer students signed a learning contract and were randomly distributed in small groups (3–4 students each). Application of this collaborative learning activity to our classrooms has been very satisfactory, as evidenced by an improvement in the volunteers’ academic performance and a very positive perception by most of them, who declared to be “very satisfied” or “satisfied” with their experience and felt that they had learned more.This work was supported by the University of Málaga (Spain) with funds granted to the educational innovation projects PIE15-163, PIE17-145, and PIE19-057. The experimental work carried out by our group is supported by grants PID2019-105010RB-I00 and EDU2017-82197-P (Spanish Ministry of Science and Innovation), UMA18-FEDERJA-220 (Andalusian Government and FEDER), as well as PY20_00257 and funds from PAIDI group BIO 267 (Andalusian Government). Funding for open access charge: Universidad de Málaga / CBU

Design and generation of a glutaminase GLS2 conditional knockout mice

Mammalian glutaminase (GA; EC 3.5.1.2) is the main enzyme involved in brain generation of glutamate (Glu). This amino acid acts as an excitatory neurotransmitter within the CNS, and it is also implicated in behavioral sensitization through the mesolimbic pathway. Two different GA genes have been described: Gls that encodes the isozymes KGA and GAC, and Gls2, which encodes GAB and LGA isozymes. Gls and Gls2 isoforms are co-expressed in different brain regions and cells. Of note, location of Gls2-encoded isoforms in neuronal nuclei suggests a novel role in the regulation of gene expression. The co-expression of different GA isoforms in mammalian brain is so far unexplained. Our objective is to study the cerebral function of Gls2; for this purpose, we develop a conditional knockout (KO) mouse model to silence GAB and LGA expression in brain. A vector carrying the Gls2 gene from exon 1 to 12 (obtained from the EUCOMM consortium) was transfected by electroporation into B6D2F1 murine embryonic stem cells (ES). These ES were selected by geneticin and PCR-genotyped before their microinjection in 8-cell stage embryos (Swiss strain). Embryo implantation was performed in pseudopregnant state mice, which leads to chimeric pups. This vector targets chromosome 10 and will yield a conditional KO mouse model, since exons 2 to 7 are included between loxP sites. The chimeric pups carrying this modification within their germ line were used to generate the homozygous Gls2 (-/-) mice. After integration of the vector in both alleles, the mice will be mated with mutant Cre mice, which express this recombinase enzyme under control of the synapsin specific promoter. This will result in a deletion of the exons 2 to 7 giving rise to null Gls2 mutants mainly in the following brain areas: cortex, hippocampus, amygdala and cerebellum, which are essential for glutamatergic transmission and related to the mesolimbic pathway.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech