Positive biocompatibility of several graphene derivatives with dopaminergic cells at long term culture

The emerging carbon nanomaterial graphene (G) and its oxidized derivative graphene oxide (GO) have recently gained considerable attention in biomedical applications such as cancer therapy or biosensors. It has for example been demonstrated that G has an efficient bioconjugation with common biomolecules and activates cell differentiation of neuronal stem cells (Li et al., 2013). This way, G could acts as a physical support or scaffold to promote axonal sprout as a “deceleration” support for the DA cells derived from neural stem cells. Since GO in its multilayer form and with multiples carboxilate and epoxy groups seems to shows interesting biological properties (Yang et al., 2013) the aim of the present work has been to test different graphene derivatives searching for the best scaffold to be used in stem cell differentiation. For this purpose we have tested the cytotoxicity of GO and reduced GO, and specifically its biocompatibility with SN4741, a dopaminergic cells line derived from mouse substance nigra, measuring the effect in the cells at long term culture. The cells were cultured in Dulbecco’s modified Eagle’s medium 10% FCS (Gibco) to about 80% confluence. Cells were incubated applying 1.000 cells in 96-well microliter plates with graphene using three chemically different types of GO as powders and films: 1) GO, which is hydrophilic; 2) partially reduced GO (PRGO) which is hydrophobic and 3) fully reduced GO (FRGO), also hydrophobic, in five concentrations: 1 mg/ml; 0.1 mg/ml; 0.05 mg/ml; 0.02 mg/ml and 0.01 mg/ml, in each type of graphene. Cells were cultured with GO and cell viability was determined after 24 hours, 1 week and 2 weeks using the MTT assay (Roche) and cytotoxicitity was determined by the lactate dehydrogenase (LDH) (Roche) assay measured at 560nm. The results demonstrated positive biocompatibility between the G-derivatives and SN4741 cells. We conclude that the use of our G-derivative scaffolds can enhance the neural differentiation towards neurons (TH positive) providing a cell growth microenvironments and appropriate synergistic cell guidance cues. This findings demonstrated that biocompatibility of scaffolds is a pre-requisite for generation of successful clinical application of graphene. It could offer a platform for neural stem cells and a promising approach for neural regeneration in the research of neurological diseases like PD. Long-term studies on the biological effects of graphene will now be performed for the development of therapeutic treatment as the goal. (Refs: Li N., Zhang Q, Gao S. et a., 2013, Nature/Sci Rep. 3:1604. doi: 10.1038/srep01604; Yan K., Li Y., Tan X., et al., 2013, Small., 9(9-10): 1492-1503)1. Universidad de Malaga. Campus de Excelencia Internacional Andalucia Tech, España. 2. The Norwegian Research Council (grant nº 215086, Oslo, Noruega. 3. Karolinska Institute Reasearch Fund, Estocolmo, Suecia

Nanocrystalline glass-like carbon thin films may be an useful tool in nerve cells regeneration

Abstract en el libro de Abstract de la International Conference, p. 99The interest in carbon nanomaterials with high transparency and electrical conductivity has grown within the last decade in view of a wide variety of applications, including biocompatible sensors, diagnostic devices and bioelectronic implants. The aim of this work is to test the biocompatibility of particular nanometer-thin nanocrystalline glass-like carbon films (NGLC), a disordered structure of graphene flakes joined by carbon matrixUniversidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. We thank Prof. Arenas, Karolinska Institutet, Stockholm, Sweden, for SN4741 cell line. R.G.V. gratefully acknowledges Spanish Ministry of Science and Innovation for funding through a Ramon y Cajal fellowship

Efecto neuroprotector de IGF-II en situaciones de estrés neuronal inducido por corticosterona

Presentado en el 15º Congreso Nacional de la SENC 2013El daño por estrés en tejido neuronal mediado por la presencia de elevados niveles de corticosterona, se relaciona con la producción de alteraciones en funciones cognitivas. Así, el estrés agudo induce un aumento importante en los niveles plasmáticos de corticosterona que se asocia con alteraciones de plasticidad neuronal. Datos previos de nuestro grupo demuestran un efecto neuroprotector a dosis bajas del factor de crecimiento similar a insulina (IGF-II) en ratas de edad avanzada, donde el estrés de tipo oxidativo se encuentra involucrado en el propio proceso senil y en el desarrollo de diferentes enfermedades neurodegenerativas. Objetivos: En el presente trabajo se pretenden estudiar las propiedades antioxidantes de dosis bajas de IGF-II en cultivos primarios de corteza neuronal de ratas adultas, sometidos a elevadas concentraciones transitorias de corticosterona. Métodos: Los parámetros de estrés oxidativo se determinaron mediante citometria de flujo y espectrofotometría. Resultados: Las neuronas corticales incubadas con corticosterona mostraron un aumento de especies reactivas de oxígeno (ROS), del consumo de reserva antioxidante celular (etax), y de la peroxidación lipídica (LOOH) todo ello corno consecuencia de un desequilibrio entre factores pro-oxidantes (ej. daño mitocondrial, NOS) y factores antioxidantes (ej. GSH, SOD). La co-incubación de corticosterona y dosis bajas de IGF-II reequilibra estos factores. Este efecto neuroprotector podría estar mediado al menos en parte por la interacción de IGF-II con sus receptores específicos, ya que su bloqueo origina de nuevo un desequilibro entre los factores antioxidantes/pro-oxidantes. Conclusión: Las dosis bajas de IGF-II ejercen un efecto neuroprotector en situaciones de estrés oxidativo inducido por altos niveles de corticosterona.Laboratorios Lilly (Madrid) por cesion del IGF-II y Ayuda de la Universidad de Malaga. Campus de Excelencia Andalucia Tec

Graphene derivative scaffolds facilitate in vitro cell survival and maturation of dopaminergic SN4741 cells

The emerging carbon nanomaterial Graphene (G), in the form of scaffold structure, has an efficient bioconjugation with common biomolecules and activates cell differentiation of neuronal stem cells, providing a promising approach for neural regeneration. We propose the use of G as a scaffold to re-address the dopaminergic (DA) neurons and the residual axons from dead or apoptotic DA neurons in Parkinson´s disease (PD). G could act as a physical support to promote the axonal sprout as a “deceleration” support for the DA cells derived from neural stem cells or DA direct cell conversion, allowing the propagation of nerve impulses. We cultured a clonal substantia nigra (SN) DA neuronal progenitor cell line (SN4741) in presence of G as scaffold. This cell line derived from mouse embryos was cultured in Dulbecco’s modified Eagle’s medium/10% FCS to about 80% confluence. Cells were incubated in three chemically different G derivatives and two different presentation matrixes as powder and films: 1) G oxide (GO); 2) partially reduced GO (PRGO) which is hydrophobic; and 3) fully reduced GO (FRGO). Cell viability was determined using the MTT assay after adding the following G concentrations: 1mg/ml; 0.1mg/ml; 0.05mg/ml; 0.02mg/ml and 0.01mg/ml, in each type of GO. To study cellular morphology and assessment of cell engraftment into GO films (GO film, PRGO film, FRGO film), we analyzed the immunostaining of the anti-rabbit neuron-specific DNA-binding protein (NeuN) antibody, the anti-rat Beta-3-tubulin antibody in combination with the mitochondrial marker mouse anti-ATP synthase antibody, and the anti-rabbit DCX as immature neuronal marker. Hoechst label was used as nuclei marker. Reactive oxidative species (ROS) were measured by flow cytometry to study the influence of G on the cell redox-state. With this purpose, cells were loaded with dihydroethidium. The mitochondrial membrane potential after JC-1 incubation was studied by flow cytometry. Our results show an increase of survival and metabolism (30-40%) at low concentrations of PRGO and FRGO (0.05-0.01 mg/ml) respect to the higher concentration (1 mg/ml), while no changes were seen in the GO group. LDH concentration was measured in the supernatant using a COBAS analyzer showing a neuroprotective action at low concentrations. Furthermore, either PRGO film or FRGO film show an increase in the effective anchorage capacity to nest into the G matrix and in the maturation of the SN 4741 cells. We conclude that the use of G scaffolds in the research of neurological diseases like PD could offer a powerful platform for neural stem cells, direct cell conversion techniques and neural tissue engineering.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. Norwegian Research Council (grant nº 215086

The graphene oxide species induce a different biological response in SN4741 Parkinson cell line

Introduction: Graphene Oxide (GO)has recently emerged as a reliable material to create scaffolds for the neural tissue because of its biocompatibility, electroconductive and physicochemical properties. Graphene is a 2-dimensional material consisting of rings of carbon atoms with an excellent electrical conductivity originating in the sp2 hybridized carbons network. Nevertheless, there is not a consensus which kind of graphene oxide is most useful of benefit.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Professional quality of life and organizational changes: a five-year observational study in Primary Care

<p>Abstract</p> <p>Background</p> <p>The satisfaction and the quality of life perceived by professionals have implications for the performance of health organizations. We have assessed the variations in professional quality of life (PQL) and their explanatory factors during a services management decentralization process.</p> <p>Methods</p> <p>It was designed as a longitudinal analytical observational study in a Health Area in Madrid, Spain. Three surveys were sent out during an ongoing management decentralization process between 2001 and 2005. The professionals surveyed were divided into three groups: Group I (97.3% physicians), group II (92.5% nurses) and group III (auxiliary personnel). Analysis of the tendency and elaboration of an explanatory multivariate model was made. The PQL -35 questionnaire, based on Karasek's demand-control theory, was used to measure PQL. This questionnaire recognizes three PQL dimensions: management support (MS), workload (WL) and intrinsic motivation (IM).</p> <p>Results</p> <p>1444 responses were analyzed. PQL increased 0.16 (CI 95% 0.04 – 0.28) points in each survey. Group II presents over time a higher PQL score than group I of 0.38 (IC 95% 0.18 – 0.59) points. There is no difference between groups I and III.</p> <p>For each point that MS increases, PQL increases between 0.44 and 0.59 points. PQL decreases an average of between 0.35 and 0.49 point, for each point that WL increases.</p> <p>Age appears to have a marginal association with PQL (CI 95% 0.00 – 0.02), as it occurs with being single or not having a stable relationship (CI 95% 0.01 – 0.41). Performing management tasks currently or in the past is related to poorer PQL perception (CI 95% -0.45 – -0.06), and the same occurs with working other than morning shifts (CI 95% -0.03 – -0.40 points).</p> <p>PQL is not related to sex, location of the centre (rural/urban), time spent working in the organization or contractual situation.</p> <p>Conclusion</p> <p>With the improvement in work control and avoiding increases in workloads, PQL perception can be maintained despite deep organizational changes at the macro-management level. Different professional groups experience different perceptions depending on how the changes impact their position in the organization.</p

The Fourteenth Data Release of the Sloan Digital Sky Survey: First Spectroscopic Data from the extended Baryon Oscillation Spectroscopic Survey and from the second phase of the Apache Point Observatory Galactic Evolution Experiment

The fourth generation of the Sloan Digital Sky Survey (SDSS-IV) has been in operation since July 2014. This paper describes the second data release from this phase, and the fourteenth from SDSS overall (making this, Data Release Fourteen or DR14). This release makes public data taken by SDSS-IV in its first two years of operation (July 2014-2016). Like all previous SDSS releases, DR14 is cumulative, including the most recent reductions and calibrations of all data taken by SDSS since the first phase began operations in 2000. New in DR14 is the first public release of data from the extended Baryon Oscillation Spectroscopic Survey (eBOSS); the first data from the second phase of the Apache Point Observatory (APO) Galactic Evolution Experiment (APOGEE-2), including stellar parameter estimates from an innovative data driven machine learning algorithm known as "The Cannon"; and almost twice as many data cubes from the Mapping Nearby Galaxies at APO (MaNGA) survey as were in the previous release (N = 2812 in total). This paper describes the location and format of the publicly available data from SDSS-IV surveys. We provide references to the important technical papers describing how these data have been taken (both targeting and observation details) and processed for scientific use. The SDSS website (www.sdss.org) has been updated for this release, and provides links to data downloads, as well as tutorials and examples of data use. SDSS-IV is planning to continue to collect astronomical data until 2020, and will be followed by SDSS-V.Comment: SDSS-IV collaboration alphabetical author data release paper. DR14 happened on 31st July 2017. 19 pages, 5 figures. Accepted by ApJS on 28th Nov 2017 (this is the "post-print" and "post-proofs" version; minor corrections only from v1, and most of errors found in proofs corrected

Estudios de Caso sobre Ciencias Agropecuarias y Rurales en el siglo XXI.

Libro científico sobre estudios de casos en el medio agropecuario y ruralCon el advenimiento del siglo XXI y el avance de los procesos de globalización, el medio rural presenta diversos cambios económicos, sociales, políticos y culturales. Lo anterior significa que el campo es un objeto de estudio altamente dinámico, complejo e inasible. las ciencias agropecuarias y rurales, en la actualidad, requieren de un abordaje sistémico e interdisciplinario que den cuenta de la heterogeneidad de situaciones y contextos que enfrenta el campo mexicano. La presente obra agrupa 18 estudios de caso, que capturan algunas fotografías de las diversas problemáticas de la ruralidad mexicana, con lo cual se pretende dar cuenta tanto de los objetivos de estudio como de la perspectiva teórico metodológico desde que estos son abordados. lo anterior tiene que ver con el hecho de que las ciencias agropecuarias y rurales manifiestan un alto grado de observación empírica, motivo por el que los estudios de caso se convierten en la perspectiva metodológica idónea que permite ir y venir de la realidad a la teoría y viceversa para la construcción de objetos de estudio. En este volumen se aborda una gran diversidad de casos, que sintetizan la heterogeneidad de enfoques y perspectivas mediante las cuales los fenómenos agropecuarios y rurales han sido abordados en el Instituto de Ciencias Agropecuarias y Rurales de la Universidad Autónoma del Estado de México, en los últimos 30 años

Hyperoxemia and excess oxygen use in early acute respiratory distress syndrome : Insights from the LUNG SAFE study

Publisher Copyright: © 2020 The Author(s). Copyright: Copyright 2020 Elsevier B.V., All rights reserved.Background: Concerns exist regarding the prevalence and impact of unnecessary oxygen use in patients with acute respiratory distress syndrome (ARDS). We examined this issue in patients with ARDS enrolled in the Large observational study to UNderstand the Global impact of Severe Acute respiratory FailurE (LUNG SAFE) study. Methods: In this secondary analysis of the LUNG SAFE study, we wished to determine the prevalence and the outcomes associated with hyperoxemia on day 1, sustained hyperoxemia, and excessive oxygen use in patients with early ARDS. Patients who fulfilled criteria of ARDS on day 1 and day 2 of acute hypoxemic respiratory failure were categorized based on the presence of hyperoxemia (PaO2 > 100 mmHg) on day 1, sustained (i.e., present on day 1 and day 2) hyperoxemia, or excessive oxygen use (FIO2 ≥ 0.60 during hyperoxemia). Results: Of 2005 patients that met the inclusion criteria, 131 (6.5%) were hypoxemic (PaO2 < 55 mmHg), 607 (30%) had hyperoxemia on day 1, and 250 (12%) had sustained hyperoxemia. Excess FIO2 use occurred in 400 (66%) out of 607 patients with hyperoxemia. Excess FIO2 use decreased from day 1 to day 2 of ARDS, with most hyperoxemic patients on day 2 receiving relatively low FIO2. Multivariate analyses found no independent relationship between day 1 hyperoxemia, sustained hyperoxemia, or excess FIO2 use and adverse clinical outcomes. Mortality was 42% in patients with excess FIO2 use, compared to 39% in a propensity-matched sample of normoxemic (PaO2 55-100 mmHg) patients (P = 0.47). Conclusions: Hyperoxemia and excess oxygen use are both prevalent in early ARDS but are most often non-sustained. No relationship was found between hyperoxemia or excessive oxygen use and patient outcome in this cohort. Trial registration: LUNG-SAFE is registered with ClinicalTrials.gov, NCT02010073publishersversionPeer reviewe

Effects of hospital facilities on patient outcomes after cancer surgery: an international, prospective, observational study

Background Early death after cancer surgery is higher in low-income and middle-income countries (LMICs) compared with in high-income countries, yet the impact of facility characteristics on early postoperative outcomes is unknown. The aim of this study was to examine the association between hospital infrastructure, resource availability, and processes on early outcomes after cancer surgery worldwide.Methods A multimethods analysis was performed as part of the GlobalSurg 3 study-a multicentre, international, prospective cohort study of patients who had surgery for breast, colorectal, or gastric cancer. The primary outcomes were 30-day mortality and 30-day major complication rates. Potentially beneficial hospital facilities were identified by variable selection to select those associated with 30-day mortality. Adjusted outcomes were determined using generalised estimating equations to account for patient characteristics and country-income group, with population stratification by hospital.Findings Between April 1, 2018, and April 23, 2019, facility-level data were collected for 9685 patients across 238 hospitals in 66 countries (91 hospitals in 20 high-income countries; 57 hospitals in 19 upper-middle-income countries; and 90 hospitals in 27 low-income to lower-middle-income countries). The availability of five hospital facilities was inversely associated with mortality: ultrasound, CT scanner, critical care unit, opioid analgesia, and oncologist. After adjustment for case-mix and country income group, hospitals with three or fewer of these facilities (62 hospitals, 1294 patients) had higher mortality compared with those with four or five (adjusted odds ratio [OR] 3.85 [95% CI 2.58-5.75]; p&lt;0.0001), with excess mortality predominantly explained by a limited capacity to rescue following the development of major complications (63.0% vs 82.7%; OR 0.35 [0.23-0.53]; p&lt;0.0001). Across LMICs, improvements in hospital facilities would prevent one to three deaths for every 100 patients undergoing surgery for cancer.Interpretation Hospitals with higher levels of infrastructure and resources have better outcomes after cancer surgery, independent of country income. Without urgent strengthening of hospital infrastructure and resources, the reductions in cancer-associated mortality associated with improved access will not be realised