Sensitivity to anti-Fas is independent of increased cathepsin D activity and adrenodoxin reductase expression occurring in NOS-3 overexpressing HepG2 cells

© 2015 Elsevier B.V. Stable overexpression of endothelial nitric oxide synthase (NOS-3) in HepG2 cells (4TO-NOS) leads to increased nitro-oxidative stress and upregulation of the cell death mediators p53 and Fas. Thus, NOS-3 overexpression has been suggested as a useful antiproliferative mechanism in hepatocarcinoma cells. We aimed to identify the underlying mechanism of cell death induced by NOS-3 overexpression at basal conditions and with anti-Fas treatment. The intracellular localization of NOS-3, the nitro-oxidative stress and the mitochondrial activity were analysed. In addition, the protein expression profile in 4TO-NOS was screened for differentially expressed proteins potentially involved in the induction of apoptosis. NOS-3 localization in the mitochondrial outer membrane was not associated with changes in the respiratory cellular capacity, but was related to the mitochondrial biogenesis increase and with a higher protein expression of mitochondrial complex IV. Nitro-oxidative stress and cell death in NOS-3 overexpressing cells occurred with the expression increase of pro-apoptotic genes and a higher expression/activity of the enzymes adrenodoxin reductase mitochondrial (AR) and cathepsin D (CatD). CatD overexpression in 4TO-NOS was related to the apoptosis induction independently of its catalytic activity. In addition, CatD activity inhibition by pepstatin A was not effective in blocking apoptosis induced by anti-Fas. In summary, NOS-3 overexpression resulted in an increased sensitivity to anti-Fas induced cell death, independently of AR expression and CatD activity.Instituto de Salud Carlos III (FIS 09/00185). G. Ferrín was supported by the Networked Biomedical Research Center Hepatic and Digestive Diseases (CIBEREHD)Peer Reviewe

OncomiRs miR-106a and miR-17 negatively regulate the nucleoside-derived drug transporter hCNT1

High-afnity uptake of natural nucleosides as well as nucleoside derivatives used in anticancer therapies is mediated by human concentrative nucleoside transporters (hCNTs). hCNT1, the hCNT family member that specifcally transports pyrimidines, is also a transceptor involved in tumor progression. In particular, oncogenesis appears to be associated with hCNT1 downregulation in some cancers, although the underlying mechanisms are largely unknown. Here, we sought to address changes in colorectal and pancreatic ductal adenocarcinoma both of which are important digestive cancers in the context of treatment with fuoropyrimidine derivatives. An analysis of cancer samples and matching non-tumoral adjacent tissues revealed downregulation of hCNT1 protein in both types of tumor. Further exploration of the putative regulation of hCNT1 by microRNAs (miRNAs), which are highly deregulated in these cancers, revealed a direct relationship between the oncomiRs miR-106a and miR-17 and the loss of hCNT1. Collectively, our fndings provide the frst demonstration that hCNT1 inhibition by these oncomiRs could contribute to chemoresistance to fuoropyrimidine-based treatments in colorectal and pancreatic cancer

Spectral Libraries for SWATH-MS Assays for Drosophila melanogaster and Solanum lycopersicum.

Quantitative proteomics methods have emerged as powerful tools for measuring protein expression changes at the proteome level. Using MS-based approaches, it is now possible to routinely quantify thousands of proteins. However, prefractionation of the samples at the protein or peptide level is usually necessary to go deep into the proteome, increasing both MS analysis time and technical variability. Recently, a new MS acquisition method named SWATH is introduced with the potential to provide good coverage of the proteome as well as a good measurement precision without prior sample fractionation. In contrast to shotgun-based MS however, a library containing experimental acquired spectra is necessary for the bioinformatics analysis of SWATH data. In this study, spectral libraries for two widely used models are built to study crop ripening or animal embryogenesis, Solanum lycopersicum (tomato) and Drosophila melanogaster, respectively. The spectral libraries comprise fragments for 5197 and 6040 proteins for S. lycopersicum and D. melanogaster, respectively, and allow reproducible quantification for thousands of peptides per MS analysis. The spectral libraries and all MS data are available in the MassIVE repository with the dataset identifiers MSV000081074 and MSV000081075 and the PRIDE repository with the dataset identifiers PXD006493 and PXD006495

Informe final del proyecto elaboración y análisis de casos psicosociales por parte del alumnado

Projecte 2014PID-UB/048A continuación se presenta el informe final del proyecto Elaboración y análisis de casos por parte del alumnado (2014PID-UB/048). Mediante este proyecto de dos años de duración se pretendió desarrollar estrategias docentes para fomentar la adquisición de la competencia específica conocer, comprender y analizar el comportamiento por parte de los alumnos de la asignatura Psicología Social del Grado de Psicología de la Universidad de Barcelona. En esta dirección, a lo largo del primer año se planificaron, desarrollaron e implementaron actividades de aprendizaje y de evaluación basadas en la metodología de estudio de casos. A lo largo del segundo año se han realizado modificaciones en las actividades inicialmente planteadas con el fin de mejorar los procesos de aprendizaje del alumnado. Los resultados obtenidos demuestran que la elaboración y el análisis de casos por parte de los estudiantes favorecen su capacidad para comprender y analizar el comportamiento humano. Asimismo, tiene efectos positivos sobre su desempeño en el examen final de la asignatura y sobre su nivel de satisfacción con el curso

Development of psychosocial case studies by students to improve their ability to understand and analyze human behavior

This study presents an active learning methodology based on the development and analysis of case studies by college students and explores its effects on academic performance and on students' capacity of understanding and analysing human behaviour. A group of 54 students who were taking the course Social Psychology at the University of Barcelona developed written stories where psychosocial concepts were represented. Results showed that participants, after developing their own case studies, improved their ability to identify and explain theoretical concepts included in case studies created by the teaching team. The proposed teaching methodology appears to enhance their ability to understand and analyse human behaviour, helping students to integrate theory with reality

Sensitivity to anti-Fas is independent of increased cathepsin D activity and adrenodoxin reductase expression occurring in NOS-3 overexpressing HepG2 cells

Stable overexpression of endothelial nitric oxide synthase (NOS-3) in HepG2 cells (4TO-NOS) leads to increased nitro-oxidative stress and upregulation of the cell death mediators p53 and Fas. Thus, NOS-3 overexpression has been suggested as a useful antiproliferative mechanism in hepatocarcinoma cells. We aimed to identify the underlying mechanism of cell death induced by NOS-3 overexpression at basal conditions and with anti-Fas treatment. The intracellular localization of NOS-3, the nitro-oxidative stress and the mitochondrial activity were analysed. In addition, the protein expression profile in 4TO-NOS was screened for differentially expressed proteins potentially involved in the induction of apoptosis. NOS-3 localization in the mitochondrial outer membrane was not associated with changes in the respiratory cellular capacity, but was related to the mitochondrial biogenesis increase and with a higher protein expression of mitochondrial complex IV. Nitro-oxidative stress and cell death in NOS-3 overexpressing cells occurred with the expression increase of pro-apoptotic genes and a higher expression/activity of the enzymes adrenodoxin reductase mitochondrial (AR) and cathepsin D (CatD). CatD overexpression in 4TO-NOS was related to the apoptosis induction independently of its catalytic activity. In addition, CatD activity inhibition by pepstatin A was not effective in blocking apoptosis induced by anti-Fas. In summary, NOS-3 overexpression resulted in an increased sensitivity to anti-Fas induced cell death, independently of AR expression and CatD activityThis study was supported by the Instituto de Salud Carlos III (FIS 09/00185). G. Ferrín was supported by the Networked Biomedical Research Center Hepatic and Digestive Diseases (CIBEREHD

Ethylene Receptors, CTRs and EIN2 Target Protein Identification and Quantification Through Parallel Reaction Monitoring During Tomato Fruit Ripening.

Ethylene, the plant ripening hormone of climacteric fruit, is perceived by ethylene receptors which is the first step in the complex ethylene signal transduction pathway. Much progress has been made in elucidating the mechanism of this pathway, but there is still a lot to be done in the proteomic quantification of the main proteins involved, particularly during fruit ripening. This work focuses on the mass spectrometry based identification and quantification of the ethylene receptors (ETRs) and the downstream components of the pathway, CTR-like proteins (CTRs) and ETHYLENE INSENSITIVE 2 (EIN2). We used tomato as a model fruit to study changes in protein abundance involved in the ethylene signal transduction during fruit ripening. In order to detect and quantify these low abundant proteins located in the membrane of the endoplasmic reticulum, we developed a workflow comprising sample fractionation and MS analysis using parallel reaction monitoring. This work shows the feasibility of the identification and absolute quantification of all seven ethylene receptors, three out of four CTRs and EIN2 in four ripening stages of tomato. In parallel, gene expression was analyzed through real-time qPCR. Correlation between transcriptomic and proteomic profiles during ripening was only observed for three of the studied proteins, suggesting that the other signaling proteins are likely post-transcriptionally regulated. Based on our quantification results we were able to show that the protein levels of SlETR3 and SlETR4 increased during ripening, probably to control ethylene sensitivity. The other receptors and CTRs showed either stable levels that could sustain, or decreasing levels that could promote fruit ripening

Recuperació de les dones en situació de violència masclista de parella : descripció i instrumentació

Podeu consultar la versió en castellà actualitzada de l'any 2016 a: http://hdl.handle.net/11703/106307Aquesta publicació presenta els resultats d'una recerca aplicada, patrocinada per l'Ajuntament de Barcelona, dirigida per la Universitat de Barcelona i realitzada amb la participació de l'Equip d'Atenció a les Dones de la Direcció del Programa de la Dona de l'Ajuntament de Barcelona i gestionat per la Fundació Salut i Comunitat

Recuperación de las mujeres en situación de violencia machista de pareja : descripción e instrumentación

Podeu consultar la versió en català de l'any 2013 a: http://hdl.handle.net/11703/91078Aquesta publicació presenta els resultats d'una recerca aplicada, patrocinada per l'Ajuntament de Barcelona, dirigida per la Universitat de Barcelona i realitzada amb la participació de l'Equip d'Atenció a les Dones de la Direcció del Programa de la Dona de l'Ajuntament de Barcelona i gestionat per la Fundació Salut i Comunitat

Striatal Blood-Brain Barrier Opening in Parkinson's Disease Dementia: A Pilot Exploratory Study

Parkinson's disease (PD) exhibits a high prevalence of dementia as disease severity and duration progress. Focused ultrasound (FUS) has been applied for transient blood-brain barrier (BBB) opening of cortical regions in neurodegenerative disorders. The striatum is a primary target for delivery of putative therapeutic agents in PD.Plan Nacional Ministerio de Educación y Ciencias (grant PID2019 111045RB-I00 to J.A.O.) and MAPFRE Foundation (J.A.O.).Peer reviewe