4 research outputs found

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

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    Presentado en el 15Âș Congreso Nacional de la SENC 2013El daño por estrĂ©s en tejido neuronal mediado por la presencia de elevados niveles de corticosterona, se relaciona con la producciĂłn de alteraciones en funciones cognitivas. AsĂ­, el estrĂ©s agudo induce un aumento importante en los niveles plasmĂĄticos 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 estrĂ©s 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. MĂ©todos: Los parĂĄmetros de estrĂ©s oxidativo se determinaron mediante citometria de flujo y espectrofotometrĂ­a. Resultados: Las neuronas corticales incubadas con corticosterona mostraron un aumento de especies reactivas de oxĂ­geno (ROS), del consumo de reserva antioxidante celular (etax), y de la peroxidaciĂłn lipĂ­dica (LOOH) todo ello corno consecuencia de un desequilibrio entre factores pro-oxidantes (ej. daño mitocondrial, NOS) y factores antioxidantes (ej. GSH, SOD). La co-incubaciĂłn de corticosterona y dosis bajas de IGF-II reequilibra estos factores. Este efecto neuroprotector podrĂ­a estar mediado al menos en parte por la interacciĂłn de IGF-II con sus receptores especĂ­ficos, ya que su bloqueo origina de nuevo un desequilibro entre los factores antioxidantes/pro-oxidantes. ConclusiĂłn: Las dosis bajas de IGF-II ejercen un efecto neuroprotector en situaciones de estrĂ©s 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

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    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 ParkinsonÂŽs 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 MĂĄlaga. Campus de Excelencia Internacional AndalucĂ­a Tech. Norwegian Research Council (grant nÂș 215086

    Acute Lung Injury Biomarkers in the Prediction of COVID-19 Severity: Total Thiol, Ferritin and Lactate Dehydrogenase.

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    SARS-CoV-2 (COVID-19) patients who develop acute respiratory distress syndrome (ARDS) can suffer acute lung injury, or even death. Early identification of severe disease is essential in order to control COVID-19 and improve prognosis. Oxidative stress (OS) appears to play an important role in COVID-19 pathogenesis; we therefore conceived a study of the potential discriminative ability of serum biomarkers in patients with ARDS and those with mild to moderate disease (non-ARDS). 60 subjects were enrolled in a single-centre, prospective cohort study of consecutively admitted patients: 29 ARDS/31 non-ARDS. Blood samples were drawn and marker levels analysed by spectrophotometry and immunoassay techniques. C-reactive protein (CRP), lactate dehydrogenase (LDH), and ferritin were significantly higher in ARDS versus non-ARDS cases at hospital admission. Leukocytes, LDH, ferritin, interleukin 6 (IL-6) and tumour necrosis factor alpha (TNF-α) were also significantly elevated in ARDS compared to non-ARDS patients during the hospital stay. Total thiol (TT) was found to be significantly lower in ARDS. Conversely, D-dimer, matrix metalloproteinase-9 (MMP-9) and advanced glycosylated end products (AGE) were elevated. Leukocytes, LDH, CRP, ferritin and IL-6 were found to be significantly higher in non-survivors. However, lymphocyte, tumour necrosis factor beta (TGF-ÎČ), and TT were lower. In summary, our results support the potential value of TT, ferritin and LDH as prognostic biomarkers for ARDS development in COVID-19 patients, distinguishing non-ARDS from ARDS (AUCs = 0.92; 0.91; 0.89) in a fast and cost-effective manner. These oxidative/inflammatory parameters appear to play an important role in COVID-19 monitoring and can be used in the clinical management of patients
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