Experiència recent en l'adaptació de la Bioquímica general de medicina a l'Espai Europeu d'Educació Superior

La Bioquímica és una materia anual del 1er any de Medicina, on els curs 2006-2007 hem tingut 431 alumnes matriculats. Els professors del 2n quatrimestre de la matèria, dedicat a l'estudi del metabolisme, hem endegat un pla per potenciar l'aprenentatge basat en l'estudiant, una de les accions proposades a la declaració de Glasgow de la EUA. El pla ha consistit en reduïr a la meitat el nombre de classes magistrals i crear sessions d'autoaprenentatge tutoritzat (SA T). Les classes magistrals s'han impartit en grans grups (100-120 alumnes) 2 cops per setmana en sessions d'una hora conduïdes per un únic professor per a tots els alumnes. Els SAT, en canvi, s'han impartit en grups petits (20 alumnes) un cop per setmana en sessions de 2 hores. Cada professor ha estat a càrrec dels mateixos 2 grups tot el quatrimestre. El format SAT parteix d'una proposta que el professor encarregat de la conferència del tema específic deixa al Campus Virtual, consistent en un seguit de qüestions de caire variat (teòric, pràctic, clínic), i relacionada tant amb aspectes tractats a la conferència com d'autoaprenentatge. Amb aquest material, els alumnes posen en comú les idees que han trovat sobre cada qüestió. La dinàmica participativa es veu afavorida pel tamany reduït de cada grup i la complicitat que s'estableix amb el professor. Atenent als resultats d'una enquesta anònima, la valoració de les SAT ha estat força positiva: de 216 alumnes que van respondre, entre el 49 i el 62%, depenent de la qüestió plantejada, han valorat molt positivament aquesta activitat, entre el 30 i el 40% l'ha valorat com neutra, i només entre el 7 i el 20% ha expressat una valoració negativa.Biochemistry is a 1st year, 2-semester compulsory subject in the UAB Medical School syllabus, where 431 students were enrolled during the course 2006-2007. The lecturers in charge of the 2nd semester, which deals with the study of metabolism, have started a new teaching plan paying more attention to active student learning, one of the items stressed in the Glasgow declaration of the EUA. The plan consisted of reducing the number of formal lectures by half, and the creation of tutorized self learning sessions (SAT). Lectures were taught to big groups (100-120 students), by different lecturers, and were scheduled twice a week in 1-hour sessions. In contrast, SAT were conducted with reduced, 20-student groups, once a week in 2-hour sessions, and each lecturer tutorized the same 2 groups during the whole semester. Students, who had to download the SAT material from the Campus Virtual site one week in advance, faced different theoretical, practical, or clinical proposals. During the SAT, they shared their previous knowledge on the subject, their bibliographic research, even their doubts and fears in a more friendly atmosphere. After the results obtained in an anonymous survey that was answered by 216 students, SAT have been very positively perceived by 49-62% of them, and only 7-20% report a negative opinion

Agonist and antagonist effects of aripiprazole on D<inf>2</inf>-like receptors controlling rat brain dopamine synthesis depend on the dopaminergic tone

Supported by grants SAF2006-08240, SAF2009-12510, and Red de Trastornos Adictivos RD06/0001/0015. G.F.M. was the recipient of a CSC fellowship.Background: The atypical antipsychotic drug aripiprazole binds with high affinity to a number of G protein coupled receptors, including dopamine D2 receptors, where its degree of efficacy as a partial agonist remains controversial. Methods: We examined the properties of aripiprazole at D2-like autoreceptors by monitoring the changes of dopamine synthesis in adult rat brain striatal minces incubated ex vivo. The effects of the dopaminergic tone on the properties of aripiprazole were assayed by comparing a basal condition (2 mM K+, low dopaminergic tone) and a stimulated condition (15 mM K+ where dopamine release mimics a relatively higher dopaminergic tone). We also used 2 reference compounds: quinpirole showed a clear agonistic activity and preclamol (S-(-)-PPP) showed partial agonism under both basal and stimulated conditions. Results: Aripiprazole under the basal condition acted as an agonist at D2-like autoreceptors and fully activated them at about 10 nM, inhibiting dopamine synthesis similarly to quinpirole. Higher concentrations of aripiprazole had effects not restricted to D2-like autoreceptor activation. Under the stimulated (15 mM K+) condition, nanomolar concentrations of aripiprazole failed to decrease dopamine synthesis but could totally block the effect of quinpirole. Conclusions: Under high dopaminergic tone, aripiprazole acts as a D2-like autoreceptor antagonist rather than as an agonist. These data show that, ex vivo, alteration of dopaminergic tone by depolarization affects the actions of aripiprazole on D2-like autoreceptors. Such unusual effects were not seen with the typical partial agonist preclamol and are consistent with the hypothesis that aripiprazole is a functionally selective D2R ligand

CHIR99021 causes inactivation of Tyrosine Hydroxylase and depletion of dopamine in rat brain striatum

Altres ajuts: acords transformatius de la UABCHIR99021, also known as laduviglusib or CT99021, is a Glycogen-synthase kinase 3β (GSK3β) inhibitor, which has been reported as a promising drug for cardiomyocyte regeneration or treatment of sensorial hearing loss. Since the activation of dopamine (DA) receptors regulates dopamine synthesis and they can signal through the β-arrestin pathway and GSK3β, we decided to check the effect of GSK3β inhibitors (CHIR99021, SB216763 and lithium ion) on the control of DA synthesis. Using ex vivo experiments with minces from rat brain striatum, we observed that CHIR99021, but not SB216763 or lithium, causes complete abrogation of both DA synthesis and accumulation, pointing to off-target effects of CHIR99021. This decrease can be attributed to tyrosine hydroxylase (TH) inhibition since the accumulation of L-DOPA in the presence of a DOPA decarboxylase inhibitor was similarly decreased. On the other hand, CHIR99021 caused a dramatic increase in the DOPAC/DA ratio, an indicator of DA metabolization, and hindered DA incorporation into striatum tissue. Tetrabenazine, an inhibitor of DA vesicular transport, also caused DA depletion and DOPAC/DA ratio increase to the same extent as CHIR99021. In addition, both CHIR99021 or SB216763, but not lithium, decreased TH phosphorylation in Ser19, but not in Ser31 or Ser40. These results demonstrate that CHIR99021 can lead to TH inactivation and DA depletion in brain striatum, opening the possibility of its use in DA-related disorders, and shows effects to be considered in future clinical trials. More work is needed to find the mechanism exerted by CHIR99021 on DA accumulation

Cellular distribution of the histamine H3 receptor in the basal ganglia : functional modulation of dopamine and glutamate neurotransmission

This is the author's version of a work that was accepted for publication in Basal ganglia. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Vol. 3 Núm. 2 (Jul. 2013)Altres ajuts: Red_de_Trastornos_Adictivos/RD06/0001/0015Histamine H3 receptors (H3R) are widely expressed in the brain where they participate in sleep-wake cycle and cognition among other functions. Despite their high expression in some regions of the basal ganglia, their functional role in this forebrain neural network remains unclear. The present findings provide in situ hybridization and immunohistochemical evidence for H3R expression in several neuronal populations of the rat basal ganglia but not in astrocytes (glial fibrillary acidic protein immunoreactive cells). We demonstrate the presence of H3R mRNA and protein in dopaminergic neurons (tyrosine hydroxylase positive) of the ventral tegmental area and substantia nigra. In the dorsal and ventral (nucleus accumbens) striatal complex we show H3R immunoreactivity in cholinergic (choline acetyltransferase immunoreactive) and GABAergic neurons (substance P, proenkephalin or dopamine D1 receptor positive) as well as in corticostriatal terminals (VGLUT1-immunoreactive). Double-labelling experiments in the medial prefrontal cortex show that H3R is expressed in D1R-positive interneurons and VGLUT1-positive corticostriatal output neurons. Our functional experiments confirm that H3R ligands modulate dopamine synthesis and the probability of glutamate release in the striatum from cortico-striatal afferents. The presence of H3R in such different neuronal populations and its involvement in the control of striatal dopaminergic and glutamatergic transmission ascribes a complex role to H3R in the function of the basal ganglia neural network

Creació de vídeos de tècniques de bioquímica i de bases de dades de preguntes comentades

El treball dut a terme s'emmarca en un objectiu general de producció de materials perquè s'utilitzin com a eines per afavorir l'autoaprenentatge en un entorn de docència semipresencial. S'han fet dos vídeos de dues tècniques experimentals fonamentals en bioquímica: la cromatografia i la fixació de radiolligands. Ambdós vídeos corresponen a la filmació d'experiments que implicaven manipulacions sofisticades i es van dur a terme en els laboratoris docents de la Unitat de Bioquímica de Medicina. S'ha confeccionat una base de dades de preguntes de resposta múltiple que consta actualment de 200 preguntes de bioquímica i biologia molecular classificades i revisades. La utilització tutoritzada dels vídeos i de les preguntes, juntament amb l'avaluació de la comprensió de sessions presencials mitjançant un test ràpid i també l'apropament del coneixement bioquímic a coneixements intuïtius habituals, han permès millorar el rendiment acadèmic de l'alumnat, ja que se n'ha incrementat el nombre que superen les assignatures i també la proporció d'alumnat amb bones qualificacions.El trabajo llevado a cabo se enmarca en un objetivo general de producción de materiales para que se utilicen como herramientas para favorecer el autoaprendizaje en un entorno de docencia semipresencial. Se han hecho dos vídeos de dos técnicas experimentales fundamentales en bioquímica: la cromatografía y la fijación de radio-ligandos. Ambos vídeos corresponden a la filmación de experimentos que implicaban manipulaciones sofisticadas y se llevaron a cabo en los laboratorios docentes de la Unidad de Bioquímica de Medicina. Se ha confeccionado una base de datos de preguntas de respuesta múltiple que consta actualmente de 200 preguntas de bioquímica y biología molecular clasificadas y revisadas. La utilización tutorizada de los vídeos y de laspreguntas, junto con la evaluación de la comprensión de sesiones presenciales mediante un test rápido y también el acercamiento del conocimiento bioquímico a conocimientos intuitivos habituales, han permitido mejorar el rendimiento académico del alumnado, ya que se ha incrementado el número que superan las asignaturas y también la proporción de alumnado con buenas calificaciones.The general objective of this project is the production of materials for use as toolsfor promoting independent learning in a semi-distance learning environment. Two videos were produced of two basic experimental techniques in Biochemistry: chromatography and radioligand binding. Both videos include the filming of experiments that involved sophisticated manipulations which had to be carried out in the teaching laboratories of the Biochemistry Unit in the Faculty of Medicine. A data-base of multiple choice questions was produced. This currently consists of 200 classified and reviewed Biochemistry and Molecular Biology questions. Supervised use of the video and the questions, together with assessment of understanding of presence sessions by means of a Rapid Test and linking biochemical knowledge to normal intuitive knowledge, has led to an improvement in the academic performance of the students, as it has increased the number of students passing the subject as well as the proportion of students with good grades

Quality of life impact of primary treatments for localized prostate cancer patients without hormonal treatment

Purpose Earlier studies evaluating the effect on quality of life (QoL) of localized prostate cancer interventions included patients receiving adjuvant hormone therapy, which could have affected their outcomes. Our objective was to compare the QoL impact of the three most common primary treatments on patients who were not receiving adjuvant hormonal treatment. PATIENTS AND METHODS: This was a prospective study of 435 patients treated with radical prostatectomy, external-beam radiotherapy, or brachytherapy. QoL was assessed before and after treatment with the Short Form-36 and the Expanded Prostate Cancer Index Composite. Differences between groups were tested by analysis of variance. Distribution of outcome at 3 years was examined by stratifying according to baseline status. Generalized estimating equation models were constructed to assess the effect of treatment over time. RESULTS: Compared with the brachytherapy group, the prostatectomy group showed greater deterioration on urinary incontinence and sexual scores but better urinary irritative-obstructive results (-18.22, -13.19, and +6.38, respectively, at 3 years; P < .001). In patients with urinary irritative-obstructive symptoms at baseline, improvement was observed in 64% of those treated with nerve-sparing radical prostatectomy. Higher bowel worsening (-2.87, P = .04) was observed in the external radiotherapy group, with 20% of patients reporting bowel symptoms. CONCLUSION: Radical prostatectomy caused urinary incontinence and sexual dysfunction but improved pre-existing urinary irritative-obstructive symptoms. External radiotherapy and brachytherapy caused urinary irritative-obstructive adverse effects and some sexual dysfunction. External radiotherapy also caused bowel adverse effects. Relevant differences between treatment groups persisted for up to 3 years of follow-up, although the difference in sexual adverse effects between brachytherapy and prostatectomy tended to decline over long-term follow-up. These results provide valuable information for clinical decision making

Spontaneous changes in brain striatal dopamine synthesis and storage dynamics ex vivo reveal end-product feedback-inhibition of tyrosine hydroxylase

Altres ajuts: acord transformatiu CRUE-CSICAltres ajuts: , The Michael J. Fox Foundation (ID15291), "la Caixa" Foundation (ID 100010434), under the agreement LCF/PR/HR17/52150003Synaptic events are important to define treatment strategies for brain disorders. In the present paper, freshly obtained rat brain striatal minces were incubated under different times and conditions to determine dopamine biosynthesis, storage, and tyrosine hydroxylase phosphorylation. Remarkably, we found that endogenous dopamine spontaneously accumulated during tissue incubation at 37 °C ex vivo while dopamine synthesis simultaneously decreased. We analyzed whether these changes in brain dopamine biosynthesis and storage were linked to dopamine feedback inhibition of its synthesis-limiting enzyme tyrosine hydroxylase. The aromatic-l-amino-acid decarboxylase inhibitor NSD-1015 prevented both effects. As expected, dopamine accumulation was increased with l-DOPA addition or VMAT2-overexpression, and dopamine synthesis decreased further with added dopamine, the VMAT2 inhibitor tetrabenazine or D2 auto-receptor activation with quinpirole, accordingly to the known synaptic effects of these treatments. Phosphorylation activation and inhibition of tyrosine hydroxylase on Ser31 and Ser40 with okadaic acid, Sp-cAMP and PD98059 also exerted the expected effects. However, no clear-cut association was found between dopamine feedback inhibition of its own biosynthesis and changes of tyrosine hydroxylase phosphorylation, assessed by Western blot and mass spectrometry. The later technique also revealed a new Thr30 phosphorylation in rat tyrosine hydroxylase. Our methodological assessment of brain dopamine synthesis and storage dynamics ex vivo could be applied to predict the in vivo effects of pharmacological interventions in animal models of dopamine-related disorders

Characterization of Novel Pathogenic Variants Leading to Caspase-8 Cleavage-Resistant RIPK1-Induced Autoinflammatory Syndrome

Pathogenic RIPK1 variants have been described as the cause of two different inborn errors of immunity. Biallelic loss-of-function variants cause the recessively inherited RIPK1 deficiency, while monoallelic variants impairing the caspase-8-mediated RIPK1 cleavage provoke a novel autoinflammatory disease (AID) called cleavage-resistant RIPK1-induced autoinflammatory (CRIA) syndrome. The aim of this study was to characterize the pathogenicity of two novel RIPK1 variants located at the cleavage site of caspase-8 detected in patients with dominantly-inherited, early-onset undefined AID. RIPK1 genotyping was performed by Sanger and next-generation sequencing. Clinical and analytical data were collected from medical charts, and in silico and in vitro assays were performed to evaluate the functional consequences. Genetic analyses identified two novel heterozygous RIPK1 variants at the caspase-8 cleavage site (p.Leu321Arg and p.Asp324Gly), which displayed a perfect intrafamilial phenotype-genotype segregation following a dominant inheritance pattern. Structural analyses suggested that these variants disrupt the normal RIPK1 structure, probably making it less accessible to and/or less cleavable by caspase-8. In vitro experiments confirmed that the p.Leu321Arg and p.Asp324Gly RIPK1 variants were resistant to caspase-8-mediated cleavage and induced a constitutive activation of necroptotic pathway in a similar manner that previously characterized RIPK1 variants causing CRIA syndrome. All these results strongly supported the pathogenicity of the two novel RIPK1 variants and the diagnosis of CRIA syndrome in all enrolled patients. Moreover, the evidences here collected expand the phenotypic and genetic diversity of this recently described AID, and provide interesting data about effectiveness of treatments that may benefit future patients

REPORT ON THE 2020 ICCAT WORKSHOP ON SMALL TUNAS BIOLOGY STUDIES FOR GROWTH AND REPRODUCTION.

SUMMARY This report describes the 2020 ICCAT workshop on small tunas biology studies for growth and reproduction, hosted by the Instituto Español de Oceanografía, Málaga, Spain. The major objectives of the workshop were: 1) starting the creation of ageing and reproduction reference sets and, 2) providing more training for the ongoing sample collection and processing to the teams involved in these studies. As approved by the SCRS in 2017, the Small Tuna Species Group intersessional meeting decided to prioritize the collection of biological samples aiming at growth, maturity and stock structure studies on three species: little tunny (Euthynnus alletteratus), Atlantic Bonito (Sarda sarda) and wahoo (Acanthocybium solandri), based on their economic importance and the lack of knowledge on their biology. This work will contribute to the next major advance in the assessment of these three species. RÉSUMÉ Le présent rapport décrit l'atelier de l’ICCAT tenu en 2020 sur les études de la biologie des thonidés mineurs pour la croissance et la reproduction, organisé par l'Instituto Español de Oceanografía, à Malaga, en Espagne. Les principaux objectifs de l'atelier étaient les suivants : 1) commencer à créer des ensembles de référence sur la détermination de l’âge et la reproduction et 2) fournir une formation plus poussée sur la collecte et le traitement des échantillons aux équipes participant à ces études. Tel qu’approuvé par le SCRS en 2017, lors de la réunion intersessions du Groupe d'espèces sur les thonidés mineurs, il a été décidé de donner la priorité à la collecte d'échantillons biologiques visant à étudier la croissance, la maturité et la structure des stocks de trois espèces : la thonine commune (Euthynnus alletteratus), la bonite à dos rayé (Sarda) et le thazard-bâtard (Acanthocybium solandri), sur la base de leur importance économique et des connaissances lacunaires sur leur biologie. Ces travaux contribueront à la prochaine grande avancée dans l'évaluation de ces trois espèces. RESUMEN Este informe describe el taller de ICCAT de 2020 sobre estudios de biología de pequeños túnidos para crecimiento y reproducción, acogido por el Instituto Español de Oceanografía en Málaga, España. Los principales objetivos del taller eran: 1) empezar la creación de conjuntos de referencia de determinación de la edad y reproducción y 2) facilitar más formación a los equipos involucrados en estos estudios para la recopilación de muestras y procesamiento en curso. Como aprobó el SCRS en 2017, en la Reunión intersesiones del Grupo de especies de pequeños túnidos se decidió priorizar la recopilación de muestras biológicas con miras a estudios de crecimiento, madurez y estructura del stock de tres especies: bacoreta (Euthynnus alletteratus), bonito (Sarda sarda) y peto (Acanthocybium solandri), basándose en su importancia económica y la falta de conocimientos sobre su biología. Este trabajo contribuirá a avanzar en la próxima evaluación de estas tres especies.Versión del edito