Role of insulin-degrading enzyme (IDE) in diabetes mellitus and insulin resistance

Insulin degrading enzyme (IDE) is a ubiquitously expressed zinc-metalloprotease that is one of the principal enzymes involved in the degradation and clearance of insulin, in addition to glucagon, amylin and the amyloid-β protein (Aβ). Several polymorphisms of the gen IDE are associated with risk for type 2 diabetes mellitus (T2DM) in humans, whereas hepatic insulin clearance is reduced in T2DM patients. Although strongly contributing to insulin clearance in vivo, the specific role of IDE in the liver -the primary site of plasma insulin clearance- remain unclear. The aim of our study is to decipher the role of IDE in hepatic insulin signaling and its impact on whole-body glucose metabolism and insulin clearance. To ablate IDE selectively in hepatocytes, mice homozygous for a floxed IDE allele were intercrossed with albumin-Cre mice (hereafter L-IDE-KO). Metabolic studies were performed in 1 and 3-months old mice to analyze plasma levels of fasting and non-fasting glucose, insulin, glucagon, triglycerides, cholesterol, amylin, Aβ-40 and their body weight. Likewise, intraperitoneal glucose, insulin and pyruvate tolerance test and hepatic plasma insulin clearance were assessed. To analyze hepatic insulin signaling, fasted mice were injected intraperitoneally with Insulin (0,75U/Kg) 10 minutes before were sacrificed. Afterwards, liver samples were examined by western blot for levels of total and phosphorilated insulin receptor (IR), protein kinase B (AKT1 and AKT2), glycogen synthase kinase 3 (GSK3), glycogen synthase (GS), forkhead box protein O1 (FoXO1) and IDE. L-IDE-KO male mice showed hyperglycaemia, glucose intolerance and insulin resistance. These changes were not associated with augmented food intake, body weight or plasma insulin levels. Surprisingly, hepatic plasma insulin clearance was similar between L-IDE-KO and WT mice. Hepatic insulin resistance in L-IDE-KO mice was associated with reduced plasma membrane IR levels (≈30%), as well as reduced phosphorylation levels (≈55%), which induced reduced phosphorylation levels of AKT1 and AKT2 by ≈45%. In summary, we have revealed a new role for IDE in the regulation of hepatic insulin signaling. We hypothesize that IDE regulates IR recycling to the plasma membrane. Finally, our data demonstrate that IDE is not essential for hepatic plasma insulin clearance.Departamento de Bioquímica y Biología Molecular y FisiologíaDoctorado en Investigación Biomédic

Estudio sobre la respuesta de Lego Mindstorms frente a diferentes lenguajes de programación

Este proyecto ha sido concebido para ayudar a futuros estudiantes, que vayan a utilizar el Lego Mindstorms NXT, a elegir e iniciarse en el lenguaje de programación que mejor se adecúe a sus necesidades. Para ello se han estudiado cuatro lenguajes de programación (NXT-G, NXC, leJOS monohilo y multihilo, y Matlab teleoperado) y se han ideado una serie de experimentos para evaluarlos. Posteriormente se ha creado una colección similar de programas en cada lenguaje, y finalmente se han comparado los resultados de todos los lenguajes para cada experimento obteniendo una serie de conclusiones. Asimismo se ha realizado una colección de videos para una mejor comprensión de los experimentos. __________________________________________________________________________________________________________________________This project has been conceived to help the prospective students, who will use the Lego Mindstorms NXT, to choose and to start learning about the programming language that fulfills their needs. There have been studied four programming languages (NXT-G, NXC, leJOS and leJOS multithreading, and teleoperated Matlab) and have been devised a list of experiments to evaluate them. After that, there has been created a similar collection of programs in each language, and finally the results of all languages have been compared for each experiment obtaining a set of conclusions. Also there has been created a video collection for a better understanding of the experiments.Ingeniería Industria

Implantación del SCE aplicando metodologías ágiles: Scrum

El presente trabajo trata adaptar una nueva metodología de trabajo, conocida como metodología ágil para la implantación en una empresa del Sistema de Control Económico de Valentín de Madariaga. La metodología sobre la que se ha trabajado durante el trabajo se trata de Scrum. Scrum nace como un método de desarrollo de software, durante los años 80 en Japón. Scrum, además de crear una forma de trabajo que mejore la eficiencia del mismo, trata de mejorar la relación entre los trabajadores, consiguiendo la superación profesional y personal de los mismos. Debido a que Scrum surge para solucionar problemas desde un punto de vista informático, a lo largo de este trabajo se ha pretendido adaptar la metodología de modo que pueda ser útil para la elaboración de los presupuestos y el análisis de la empresa. De este modo habrá actividades que Scrum sugiera que no sean estrictamente necesarios para la implantación que se pretende realizar. La realización de este trabajo muestra la gran capacidad de adaptación que pueden tener dichas metodologías, las cuales pueden llegar a ser realmente útiles, sobre todo a la hora de plantear y organizar actividades de cualquier tipo. Además, la implantación del SCE en la empresa ha permitido que dicha empresa mejore su producción durante el comienzo del mes de junio mediante la identificación de los defectos en la producción, así como la contabilización económica de dichos problemas.The present project tries to adapt a new work methodology, known as agile methodology to the implantation in a company of the Economic Control System developed by Valentín de Madariaga. The methodology that has been worked on during the project is called Scrum. Scrum was born as a method of software development, during the 80s in Japan. Scrum, in addition to creating a way of working that improves the efficiency of it, tries to improve the relationship between the workers, achieving the professional and personal improvement of them. Due to the Scrum arises to solve problems from a computer point of view, throughout this work it has been tried to adapt the methodology so that it can be useful for the elaboration of the budgets and the analysis of the company. In this way there will be activities that Scrum suggests that are not strictly necessary for the implementation that is intended to be carried out. The realization of this work shows the great adaptability that these methodologies can have, which can be really useful, especially when planning and organizing activities of any kind. In addition, the implementation of the SCE in the company has allowed this company to improve its production during the beginning of June by identifying the defects in the production as well as the economic accounting of these problems.Universidad de Sevilla. Grado en Ingeniería de las Tecnologías Industriale

Identification of NRF2 activation as a prognostic biomarker in T-cell acute lymphoblastic leukaemia

The standard-of-care treatment of T-cell acute lymphoblastic leukaemia (T-ALL) with chemotherapy usually achieves reasonable rates of initial complete response. However, patients who relapse or do not respond to conventional therapy show dismal outcomes, with cure rates below 10% and limited therapeutic options. To ameliorate the clinical management of these patients, it is urgent to identify biomarkers able to predict their outcomes. In this work, we investigate whether NRF2 activation constitutes a biomarker with prognostic value in T-ALL. Using transcriptomic, genomic, and clinical data, we found that T-ALL patients with high NFE2L2 levels had shorter overall survival. Our results demonstrate that the PI3K-AKT-mTOR pathway is involved in the oncogenic signalling induced by NRF2 in T-ALL. Furthermore, T-ALL patients with high NFE2L2 levels displayed genetic programs of drug resistance that may be provided by NRF2-induced biosynthesis of glutathione. Altogether, our results indicate that high levels of NFE2L2 may be a predictive biomarker of poor treatment response in T-ALL patients, which would explain the poor prognosis associated with these patients. This enhanced understanding of NRF2 biology in T-ALL may allow a more refined stratification of patients and the proposal of targeted therapies, with the ultimate goal of improving the outcome of relapsed/refractory T-ALL patients.RTI2018-093330-B-I0

Fracturas del extremo distal del radio en el esqueleto inmaduro: estudio epidemiológico en el Centro Hospitalario Pereira Rossell

Objetivo principal: Conocer la epidemiologia de las fracturas del cuarto distal del radio en el esqueleto en crecimiento y el tratamiento realizado en el CHPR en los años 2017 y 2018. Objetivos específicos: Valorar re-desplazamiento, necesidad de re manipulación, complicaciones, re-fractura. Metodología: Estudio observacional descriptivo retrospectivo. Criterios de inclusión: pacientes de 0 a 14 años con fractura de radio distal (fisaria, metafisaria y suprametafisaria) valorados en el CHPR entre 1 enero del 2017 y 31 diciembre del 2018. Criterios de exclusión: pacientes con radiografía normal, fracturas en miembros con malformaciones, infecciones óseas, patología tumoral maligna o benigna. Obtención de datos: Valoración de radiografías de puño y antebrazo realizadas en el CHPR en 2017 y 18, iniciales y evolutivas. Se analizaron: edad, sexo, fecha fractura, topografía de la fractura, desplazamiento inicial, tratamiento, evolución radiográfica y complicaciones. Resultados: se incluyeron 662 pacientes. Siendo en su mayoría de género masculino (65%), con una media de 9 años, miembro derecho (61%), en los meses de verano (36%). En cuanto a la topografía se evidenció una frecuencia mayor en fracturas tipo rodete y metafisarias (31.72% y 31.57%), seguido por suprametafisaria (18.43%) y fisaria (18.28%). El tratamiento realizado fue ortopédico en el 86.56% de los casos, mientras que quirúrgico fue el 12.84%, en su gran mayoría con alambres de Kirschner (11,2%). El tiempo de inmovilización promedio fue de 6 semanas, con un porcentaje de complicaciones del 14.05% del total de las fracturas. Conclusiones: Se valoraron las características de los pacientes y fracturas de radio distal en el CHPR en los años 2017 y 18, siendo un total de 662 fracturas, en las que su mayoría se realizó tratamiento ortopédico con un índice de complicaciones que ronda el 14%, siendo mayor cuanto mayor es el desplazamiento inicial de la fractura

Leptolide improves insulin resistance in diet-induced obese mice

Producción CientíficaType 2 diabetes (T2DM) is a complex disease linked to pancreatic beta-cell failure and insulin resistance. Current antidiabetic treatment regimens for T2DM include insulin sensitizers and insulin secretagogues. We have previously demonstrated that leptolide, a member of the furanocembranolides family, promotes pancreatic beta-cell proliferation in mice. Considering the beneficial effects of leptolide in diabetic mice, in this study, we aimed to address the capability of leptolide to improve insulin resistance associated with the pathology of obesity. To this end, we tested the hypothesis that leptolide should protect against fatty acid-induced insulin resistance in hepatocytes. In a time-dependent manner, leptolide (0.1 µM) augmented insulin-stimulated phosphorylation of protein kinase B (PKB) by two-fold above vehicle-treated HepG2 cells. In addition, leptolide (0.1 µM) counteracted palmitate-induced insulin resistance by augmenting by four-fold insulin-stimulated phosphorylation of PKB in HepG2 cells. In vivo, acute intraperitoneal administration of leptolide (0.1 mg/kg and 1 mg/kg) improved glucose tolerance and insulin sensitivity in lean mice. Likewise, prolonged leptolide treatment (0.1 mg/kg) in diet-induced obese mice improved insulin sensitivity. These effects were paralleled with an ~50% increased of insulin-stimulated phosphorylation of PKB in liver and skeletal muscle and reduced circulating pro-inflammatory cytokines in obese mice. We concluded that leptolide significantly improves insulin sensitivity in vitro and in obese mice, suggesting that leptolide may be another potential treatment for T2DM.This research has been funded by Sociedad Española de Diabetes (Ayudas Investigación Básica 2014), Salud Castilla y León (BIO/VA40/15)Ministerio de Economía y Competitividad, (SAF2014-58702-C2-1-R),(SAF2014-58702-C2-2-R

Protective effects of epoxypukalide on pancreatic b-cells and glucose metabolism in STZ-induced diabetic mice

Producción CientíficaDiabetes is a consequence of a decrease on functional β-cell mass. We have recently demonstrated that epoxypukalide (Epoxy) is a natural compound with beneficial effects on primary cultures of rat islets. In this study, we extend our previous investigations to test the hypothesis that Epoxy protects β-cells and improves glucose metabolism in STZ-induced diabetic mice. We used 3-months old male mice that were treated with Epoxy at 200 μg/kg body weight. Glucose intolerance was induced by multiple intraperitoneal low-doses of streptozotocin (STZ) on 5 consecutive days. Glucose homeostasis was evaluated measuring plasma insulin levels and glucose tolerance. Histomorphometry was used to quantify the number of pancreatic β-cells per islet. β-cell proliferation was assessed by BrdU incorporation, and apoptosis by TUNEL staining. Epoxy treatment significantly improved glucose tolerance and plasma insulin levels. These metabolic changes were associated with increased β-cell numbers, as a result of a two-fold increase in β-cell proliferation and a 50% decrease in β-cell death. Our results demonstrate that Epoxy improves whole-body glucose homeostasis by preventing pancreatic β-cell death due to STZ-induced toxicity in STZ-treated mic

Leptolide improves insulin resistance in diet-induced obese mice

Type 2 diabetes (T2DM) is a complex disease linked to pancreatic beta-cell failure and insulin resistance. Current antidiabetic treatment regimens for T2DM include insulin sensitizers and insulin secretagogues. We have previously demonstrated that leptolide, a member of the furanocembranolides family, promotes pancreatic beta-cell proliferation in mice. Considering the beneficial effects of leptolide in diabetic mice, in this study, we aimed to address the capability of leptolide to improve insulin resistance associated with the pathology of obesity. To this end, we tested the hypothesis that leptolide should protect against fatty acid-induced insulin resistance in hepatocytes. In a time-dependent manner, leptolide (0.1 µM) augmented insulin-stimulated phosphorylation of protein kinase B (PKB) by two-fold above vehicle-treated HepG2 cells. In addition, leptolide (0.1 µM) counteracted palmitate-induced insulin resistance by augmenting by four-fold insulin-stimulated phosphorylation of PKB in HepG2 cells. In vivo, acute intraperitoneal administration of leptolide (0.1 mg/kg and 1 mg/kg) improved glucose tolerance and insulin sensitivity in lean mice. Likewise, prolonged leptolide treatment (0.1 mg/kg) in diet-induced obese mice improved insulin sensitivity. These effects were paralleled with an ~50% increased of insulin-stimulated phosphorylation of PKB in liver and skeletal muscle and reduced circulating pro-inflammatory cytokines in obese mice. We concluded that leptolide significantly improves insulin sensitivity in vitro and in obese mice, suggesting that leptolide may be another potential treatment for T2DMSociedad Española de Diabetes (Ayudas Investigación Básica 2014), Salud Castilla y León (BIO/VA40/15) and Ministerio de Economía y Competitividad-Spain (SAF2014-58702-C2-1-R) to I.C. and Ministerio de Economía y Competitividad-Spain (SAF2014-58702-C2-2-R) to G.P

A Quick Guide for Using Microsoft Onenote as an Electronic Laboratory Notebook

[Abstract] Scientific data recording and reporting systems are of a great interest for endorsing reproducibility and transparency practices among the scientific community. Current research generates large datasets that can no longer be documented using paper lab notebooks (PLNs). In this regard, electronic laboratory notebooks (ELNs) could be a promising solution to replace PLNs and promote scientific reproducibility and transparency. We previously analyzed five ELNs and performed two survey-based studies to implement an ELN in a biomedical research institute. Among the ELNs tested, we found that Microsoft OneNote presents numerous features related to ELN best functionalities. In addition, both surveyed groups preferred OneNote over a scientifically designed ELN (PerkinElmer Elements). However, OneNote remains a general note-taking application and has not been designed for scientific purposes. We therefore provide a quick guide to adapt OneNote to an ELN workflow that can also be adjusted to other nonscientific ELNs

iPSC-Based Modeling of Variable Clinical Presentation in Hypertrophic Cardiomyopathy.

BACKGROUND Hypertrophic cardiomyopathy (HCM) is the most common inherited cardiac disease and a frequent cause of heart failure and sudden cardiac death. Our understanding of the genetic bases and pathogenic mechanisms underlying HCM has improved significantly in the recent past, but the combined effect of various pathogenic gene variants and the influence of genetic modifiers in disease manifestation are very poorly understood. Here, we set out to investigate genotype-phenotype relationships in 2 siblings with an extensive family history of HCM, both carrying a pathogenic truncating variant in the MYBPC3 gene (p.Lys600Asnfs*2), but who exhibited highly divergent clinical manifestations. METHODS We used a combination of induced pluripotent stem cell (iPSC)-based disease modeling and CRISPR (clustered regularly interspersed short palindromic repeats)/Cas9 (CRISPR-associated protein 9)-mediated genome editing to generate patient-specific cardiomyocytes (iPSC-CMs) and isogenic controls lacking the pathogenic MYBPC3 variant. RESULTS Mutant iPSC-CMs developed impaired mitochondrial bioenergetics, which was dependent on the presence of the mutation. Moreover, we could detect altered excitation-contraction coupling in iPSC-CMs from the severely affected individual. The pathogenic MYBPC3 variant was found to be necessary, but not sufficient, to induce iPSC-CM hyperexcitability, suggesting the presence of additional genetic modifiers. Whole-exome sequencing of the mutant carriers identified a variant of unknown significance in the MYH7 gene (p.Ile1927Phe) uniquely present in the individual with severe HCM. We finally assessed the pathogenicity of this variant of unknown significance by functionally evaluating iPSC-CMs after editing the variant. CONCLUSIONS Our results indicate that the p.Ile1927Phe variant of unknown significance in MYH7 can be considered as a modifier of HCM expressivity when found in combination with truncating variants in MYBPC3. Overall, our studies show that iPSC-based modeling of clinically discordant subjects provides a unique platform to functionally assess the effect of genetic modifiers.The funding for this research was provided by the Spanish Ministry of Science and Innovation-MCIN (grants PID2021-123925OB-I00, PID2019-104776RB-I00, CB06/01/1056, and CB16/11/00399 financed by MCIN/AEI/10.13039/501100011033), AGAUR (2021-SGR-974), Fundació La Marató de TV3 (201534-30), Fundación BBVA (BIO14_298), Fundació Obra Social la Caixa, and CERCA Program/ Generalitat de Catalunya. The CNIC is supported by the Instituto de Salud Carlos III (ISCIII), the MCIN, and the Pro CNIC Foundation. I. Lazis was partially supported by a predoctoral fellowship from MCIN (PRE2019-087901).S