SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding

Background: Protein phosphatase 2A (PP2A) is a family of conserved serine/threonine phosphatases involved in several essential aspects of cell growth and proliferation. PP2A(Cdc55) phosphatase has been extensively related to cell cycle events in budding yeast; however, few PP2A(Cdc55) substrates have been identified. Here, we performed a quantitative mass spectrometry approach to reveal new substrates of PP2A(Cdc55) phosphatase and new PP2A-related processes in mitotic arrested cells. Results: We identified 62 statistically significant PP2A(Cdc55) substrates involved mainly in actin-cytoskeleton organization. In addition, we validated new PP2A(Cdc55) substrates such as Slk19 and Lte1, involved in early and late anaphase pathways, and Zeo1, a component of the cell wall integrity pathway. Finally, we constructed docking models of Cdc55 and its substrate Mob1. We found that the predominant interface on Cdc55 is mediated by a protruding loop consisting of residues 84-90, thus highlighting the relevance of these aminoacids for substrate interaction. Conclusions: We used phosphoproteomics of Cdc55-deficient cells to uncover new PP2A(Cdc55) substrates and functions in mitosis. As expected, several hyperphosphorylated proteins corresponded to Cdk1-dependent substrates, although other kinases' consensus motifs were also enriched in our dataset, suggesting that PP2A(Cdc55) counteracts and regulates other kinases distinct from Cdk1. Indeed, Pkc1 emerged as a novel node of PP2A(Cdc55) regulation, highlighting a major role of PP2A(Cdc55) in actin cytoskeleton and cytokinesis, gene ontology terms significantly enriched in the PP2A(Cdc55)-dependent phosphoproteome

Validation of three predictive models for suboptimal cytoreductive surgery in advanced ovarian cancer

The standard treatment for advanced ovarian cancer (AOC) is cytoreduction surgery and adjuvant chemotherapy. Tumor volume after surgery is a major prognostic factor for these patients. The ability to perform complete cytoreduction depends on the extent of disease and the skills of the surgical team. Several predictive models have been proposed to evaluate the possibility of performing complete cytoreductive surgery (CCS). External validation of the prognostic value of three predictive models (Fagotti index and the R3 and R4 models) for predicting suboptimal cytoreductive surgery (SCS) in AOC was performed in this study. The scores of the 3 models were evaluated in one hundred and three consecutive patients diagnosed with AOC treated in a tertiary hospital were evaluated. Clinicopathological features were collected prospectively and analyzed retrospectively. The performance of the three models was evaluated, and calibration and discrimination were analyzed. The calibration of the Fagotti, R3 and R4 models showed odds ratios of obtaining SCSs of 1.5, 2.4 and 2.4, respectively, indicating good calibration. The discrimination of the Fagotti, R3 and R4 models showed an area under the ROC curve of 83%, 70% and 81%, respectively. The negative predictive values of the three models were higher than the positive predictive values for SCS. The three models were able to predict suboptimal cytoreductive surgery for advanced ovarian cancer, but they were more reliable for predicting CCS. The R4 model discriminated better because it includes the laparotomic evaluation of the peritoneal carcinomatosis index

SILAC-based phosphoproteomics reveals new PP2A-Cdc55-regulated processes in budding yeast

18 páginas, 5 figuras, 1 tablaBackground: Protein phosphatase 2A (PP2A) is a family of conserved serine/threonine phosphatases involved in several essential aspects of cell growth and proliferation. PP2ACdc55 phosphatase has been extensively related to cell cycle events in budding yeast; however, few PP2ACdc55 substrates have been identified. Here, we performed a quantitative mass spectrometry approach to reveal new substrates of PP2ACdc55 phosphatase and new PP2A-related processes in mitotic arrested cells. Results: We identified 62 statistically significant PP2ACdc55 substrates involved mainly in actin-cytoskeleton organization. In addition, we validated new PP2ACdc55 substrates such as Slk19 and Lte1, involved in early and late anaphase pathways, and Zeo1, a component of the cell wall integrity pathway. Finally, we constructed docking models of Cdc55 and its substrate Mob1. We found that the predominant interface on Cdc55 is mediated by a protruding loop consisting of residues 84-90, thus highlighting the relevance of these aminoacids for substrate interaction. Conclusions: We used phosphoproteomics of Cdc55-deficient cells to uncover new PP2ACdc55 substrates and functions in mitosis. As expected, several hyperphosphorylated proteins corresponded to Cdk1-dependent substrates, although other kinases' consensus motifs were also enriched in our dataset, suggesting that PP2ACdc55 counteracts and regulates other kinases distinct from Cdk1. Indeed, Pkc1 emerged as a novel node of PP2ACdc55 regulation, highlighting a major role of PP2ACdc55 in actin cytoskeleton and cytokinesis, gene ontology terms significantly enriched in the PP2ACdc55-dependent phosphoproteome.Work in our laboratory is supported by the Spanish Ministry of Science and Innovation (BFU2011–27568), Spanish Ministry of Economy and Competitively (BFU2013–43132-P and BFU2016–77975-R AEI/FEDER, UE cofounded by FEDER funds/European Regional Development Fund- a way to build Europe). MRL was supported by the Lundbeck foundation (Junior Group Leader Fellowship). This work was supported by a generous grant from the VILLUM Foundation to the VILLUM Centre for Bioanalytical Sciences at the University of Southern Denmark. SBB is a recipient of ISCIII grant 13FIS037. IDIBELL Proteomics Unit belongs to ProteoRed, PRB2-ISCIII, and is supported by grant PT13/0001/0033.Peer reviewe

NGAL release from peripheral blood mononuclear cells protects against acute kidney injury and prevents AKI induced fibrosis

We propose the use of a peripheral blood mononuclear cell therapy based on cell NGAL release to be used in the clinical setting for acute kidney injury (AKI) and the derived fibrosis. First, we designed a procedure whereby PBMC overexpress NGAL and anti-inflammatory agents when subjected to repetitive anoxia/reoxygenation (PBMC (A/R)). Using an in vivo AKI model, we observed that PBMC(A/R) reduces BUN and creatinine levels in blood and inflammation, enhances anti-inflammation, induces proliferation of tubular epithelial cells and reduces AKI-induced fibrosis. Flow cytometry analysis evidenced that monocytes are the only cells accumulated in the injured kidney and phenotype analysis of freshly isolated kidney macrophages, revealed that the healing phenotype is maintained the time needed for recovery. NGAL release from PBMC(A/R) determines the beneficial effect of the therapy since administration of a NGAL antibody previous to the therapy or injection of PBMC(A/R) obtained from NGAL KO animals abolished the beneficial effects. CD11b–NGAL positive cells were enhanced in tissue after PBMC (A/R) therapy and were produced by the injected monocytes. In an in vitro model with tubular epithelial cells (NRK52e) we proved that NGAL release by PBMC(A/R) induced epithelial proliferation and activation of PI3K/Akt pathway.This research was funded by grants RTC2019-0079197-1 funded by Ministerio de Ciencia e Innovacion/Agencia ´ Estatal de Investigacion ´ MCIN/AEI/10.13039/501100011033, awarded to G.H and grant PI20/ 00900 funded by Fondo de Investigacion ´ Sanitaria (FIS), awarded to G. H

La situación del turismo comunitario en Ecuador

Este estudio se realizó en 36 Comunidades y/o Centros de Turismo Comunitario (CTC) del Ecuador, el mismo que da cuenta sobre el impacto de esta actividad en el ámbito social, ambiental y de sostenibilidad de los emprendimientos relacionados con el sector, donde se encuentra un segmento de la población más vulnerable de la sociedad ecuatoriana, como son los habitantes de las áreas rurales. También se analizó la situación real del turismo y su influencia social, económica y ambiental como rama de la actividad dedicada a la prestación de los servicios y atención a los visitantes que buscan conseguir la satisfacción de las necesidades espirituales, de conocimiento, recreación, descanso personal y familiar. El estudio evidencia los aportes positivos del Turismo Comunitario en Ecuador, como son la generación de puestos de trabajo, el posicionamiento en mercados no tradicionales, la utilización racional de los recursos naturales, bases fundamentales del turismo ecológico y la responsabilidad social – comunitaria a partir de la organización comunitaria, la identificación de los recursos disponibles para el turismo; así como se evidencian algunas debilidades en cuanto a la disponibilidad de la infraestructura básica, promoción y la deficiente organización laboral en el sector público

Evaluación económico-ecológica de temas de investigación agropecuaria en los Países Andinos

Dentro de los objetivos para la Subregión Andina se elaboró un Plan de Trabajo con los siguientes objetivos: a. capacitar a técnicos y directivos de INIAs y otras instituciones de investigación agropecuaria en metodologías y aplicaciones de priorización; b. fortalecer sistemas de información y bases de datos de prioridades en el nivel subregional; c. desarrollar y aplicar metodologías de priorización de la investigación multinacional y nacional