Study of protein expresion [sic] in peri-infarct tissue after cerebral ischemia

In this work, we report our study of protein expression in rat peri-infarct tissue, 48 h after the induction of permanent focal cerebral ischemia. Two proteomic approaches, gel electrophoresis with mass spectrometry and combined fractional diagonal chromatography (COFRADIC), were performed using tissue samples from the periphery of the induced cerebral ischemic lesions, using tissue from the contra-lateral hemisphere as a control. Several protein spots (3408) were identified by gel electrophoresis, and 11 showed significant differences in expression between peri-infarct and contralateral tissues (at least 3-fold, p < 0.05). Using COFRADIC, 5412 proteins were identified, with 72 showing a difference in expression. Apart from blood-related proteins (such as serum albumin), both techniques showed that the 70 kDa family of heat shock proteins were highly expressed in the peri-infarct tissue. Further studies by 1D and 2D western blotting and immunohistochemistry revealed that only one member of this family (the inducible form, HSP72 or HSP70i) is specifically expressed by the peri-infarct tissue, while the majority of this family (the constitutive form, HSC70 or HSP70c) is expressed in the whole brain. Our data support that HSP72 is a suitable biomarker of peri-infarct tissue in the ischemic brain

Corporate social responsibility practices in the hotel sector. Case studies in Santa Marta city, Colombia

[ES] El presente trabajo tiene como fin identificar, diagnosticar y evaluar las prácticas de Responsabilidad Social Empresarial (RSE) de ocho hoteles de diferentes tamaños ubicados en la ciudad de Santa Marta, Colombia. A partir de la teoría del desarrollo sostenible fundamentada en la ética, fue aplicado el método del estudio de caso, para concluir que las empresas hoteleras se ubican dentro de un rango de aplicación mínimo, bajo y medio. De igual manera, se encuentran clasificadas en los grupos de reactivas-filantrópicas, reactivas-legales y activas, dando una mayor importancia y valoración al desarrollo de actividades económicas relacionadas con clientes, producto, trabajadores y proveedores locales. En relación a las implicaciones, el estudio brinda información útil en materia de RSE, para mejorar la toma decisiones, tanto de los propietarios de los hoteles, como de sus stakeholders más influyentes. En concreto, para el empresario, en gestión y operación; para el dirigente político, en política pública del sector; para el cliente, en decisiones de compra; para la academia, en decisiones sobre investigaciones en marcha o nuevas investigaciones; y para la comunidad local, en control y seguimiento que permita legitimar o no la actividad empresarial. Futuras investigaciones deberían incluir más stakeholders y empresas de otros sectores económicos de la zona del estudio.[EN] This paper aims to identify, diagnose and evaluate the practices of Corporate Social Responsibility (CSR) of eight hotels of different sizes located in Santa Marta, Colombia. From the theory of sustainable development based on ethic, the case study method was applied. It was found that hotel companies are located within a range of minimum, low and medium application, classified in philanthropic-reactive, legal-reactive and active groups, giving greater importance and value to the development of economic activities related to customers, product, workers and local suppliers. The study provides useful information on CSR, to improve decision making, both hotel owners, and their most influential stakeholders. Specifically, for employers, management and operation; for political leaders, public policy in the sector; for customers, in purchasing decisions; for academy, in decisions about ongoing investigations or further research; for local community, enabling control and monitoring legitimize or business. Future research should include more stakeholders and companies from other economic sectors in the study area.Este artículo es resultado de la Pasantía de Investigación en el marco del Programa de Doctorado Interuniversitario en Turismo de la Universidad de Málaga del primer autor en el Grupo de Investigación en Sistemas Costeros (COL 087006) de la compañía Playas Corporación Ltd. (Santa Marta, Colombia)

Performance analysis of massively parallel embedded hardware architectures for retinal image processing

This paper examines the implementation of a retinal vessel tree extraction technique on different hardware platforms and architectures. Retinal vessel tree extraction is a representative application of those found in the domain of medical image processing. The low signal-to-noise ratio of the images leads to a large amount of low-level tasks in order to meet the accuracy requirements. In some applications, this might compromise computing speed. This paper is focused on the assessment of the performance of a retinal vessel tree extraction method on different hardware platforms. In particular, the retinal vessel tree extraction method is mapped onto a massively parallel SIMD (MP-SIMD) chip, a massively parallel processor array (MPPA) and onto an field-programmable gate arrays (FPGA)This work is funded by Xunta de Galicia under the projects 10PXIB206168PR and 10PXIB206037PR and the program Maria BarbeitoS

Glutamate Excitoxicity Is the Key Molecular Mechanism Which Is Influenced by Body Temperature during the Acute Phase of Brain Stroke

Glutamate excitotoxicity, metabolic rate and inflammatory response have been associated to the deleterious effects of temperature during the acute phase of stroke. So far, the association of temperature with these mechanisms has been studied individually. However, the simultaneous study of the influence of temperature on these mechanisms is necessary to clarify their contributions to temperature-mediated ischemic damage. We used non-invasive Magnetic Resonance Spectroscopy to simultaneously measure temperature, glutamate excitotoxicity and metabolic rate in the brain in animal models of ischemia. The immune response to ischemia was measured through molecular serum markers in peripheral blood. We submitted groups of animals to different experimental conditions (hypothermia at 33°C, normothermia at 37°C and hyperthermia at 39°C), and combined these conditions with pharmacological modulation of glutamate levels in the brain through systemic injections of glutamate and oxaloacetate. We show that pharmacological modulation of glutamate levels can neutralize the deleterious effects of hyperthermia and the beneficial effects of hypothermia, however the analysis of the inflammatory response and metabolic rate, demonstrated that their effects on ischemic damage are less critical than glutamate excitotoxity. We conclude that glutamate excitotoxicity is the key molecular mechanism which is influenced by body temperature during the acute phase of brain stroke

Spatio-temporal profile, phenotypic diversity, and fate of recruited monocytes into the post-ischemic brain

Infiltrating CX3CR1GFP/+ and CCR2RFP/+ cells are monocyte/macrophages. (A) Representative fluorescent merged images and orthogonal views of CX3CR1GFP/+ (green) and Iba1+ (red) ramified (upper panels) and amoeboid cells (lower panels) showing co-localization of both markers. The images were taken from the ischemic hemisphere at 14 days post-MCAo. Scale bar represent 20 μm. (B) Representative fluorescent merged images and orthogonal views of co-localization for the markers CCR2GFP/+ (red) and Iba1+ (blue). The images were taken from the ischemic hemisphere at 3 days post-MCAo. Scale bar represent 20 μm. (PDF 10117 kb

Perturbation theory/machine learning model of ChEMBL data for dopamine targets: docking, synthesis, and assay of new l-prolyl-l-leucyl-glycinamide peptidomimetics

[Abstract] Predicting drug–protein interactions (DPIs) for target proteins involved in dopamine pathways is a very important goal in medicinal chemistry. We can tackle this problem using Molecular Docking or Machine Learning (ML) models for one specific protein. Unfortunately, these models fail to account for large and complex big data sets of preclinical assays reported in public databases. This includes multiple conditions of assays, such as different experimental parameters, biological assays, target proteins, cell lines, organism of the target, or organism of assay. On the other hand, perturbation theory (PT) models allow us to predict the properties of a query compound or molecular system in experimental assays with multiple boundary conditions based on a previously known case of reference. In this work, we report the first PTML (PT + ML) study of a large ChEMBL data set of preclinical assays of compounds targeting dopamine pathway proteins. The best PTML model found predicts 50000 cases with accuracy of 70–91% in training and external validation series. We also compared the linear PTML model with alternative PTML models trained with multiple nonlinear methods (artificial neural network (ANN), Random Forest, Deep Learning, etc.). Some of the nonlinear methods outperform the linear model but at the cost of a notable increment of the complexity of the model. We illustrated the practical use of the new model with a proof-of-concept theoretical–experimental study. We reported for the first time the organic synthesis, chemical characterization, and pharmacological assay of a new series of l-prolyl-l-leucyl-glycinamide (PLG) peptidomimetic compounds. In addition, we performed a molecular docking study for some of these compounds with the software Vina AutoDock. The work ends with a PTML model predictive study of the outcomes of the new compounds in a large number of assays. Therefore, this study offers a new computational methodology for predicting the outcome for any compound in new assays. This PTML method focuses on the prediction with a simple linear model of multiple pharmacological parameters (IC50, EC50, Ki, etc.) for compounds in assays involving different cell lines used, organisms of the protein target, or organism of assay for proteins in the dopamine pathway.Ministerio de Economía y Competitividad; CTQ2016-74881-PGobierno Vasco; IT1045-16Xunta de Galicia; GPC2014/003Xunta de Galicia; CN 2012/069Xunta de Galicia; ED431D 2017/16Xunta de Galicia; ED431D 2017/23Xunta de Galicia; GRC2014/049Xunta de Galicia; ED431D 2017/2

Association between neuroserpin and molecular markers of brain damage in patients with acute ischemic stroke.

BACKGROUND: Neuroserpin has shown neuroprotective effects in animal models of cerebral ischemia and has been associated with functional outcome after ischemic stroke. Our aim was to study whether neuroserpin serum levels could be associated to biomarkers of excitotoxicity, inflammation and blood brain barrier disruption. METHODS: We prospectively included 129 patients with ischemic stroke (58.1% male; mean age, 72.4 ± 9.6 years) not treated with tPA within 12 hours (h) of symptoms onset (mean time, 4.7 ± 2.1 h). Poor functional outcome at 3 months was considered as a modified Rankin scale score >2. Serum levels of neuroserpin, Interleukin 6 (IL-6), Intercellular adhesion molecule-1 (ICAM-1), active Matrix metalloproteinase 9 (MMP-9), and cellular fibronectin (cFn) (determined by ELISA) and glutamate (determined by HPLC) were measured on admission, 24 and 72 h. The main variable was considered the decrease of neuroserpin levels within the first 24 h. ROC analysis was used to select the best predictive value for neuroserpin to predict poor functional outcome due to a lack of linearity. RESULTS: The decrease of neuroserpin levels within the first 24 h was negatively correlated with serum levels at 24 hours of glutamate (r = -0.642), IL-6 (r = -0.678), ICAM-1 (r = -0.345), MMP-9 (r = -0.554) and cFn (r = -0.703) (all P < 0.0001). In the multivariate analysis, serum levels of glutamate (OR, 1.04; CI95%, 1.01-1.06, p = 0.001); IL-6 (OR, 1.4; CI95%, 1.1-1.7, p = 0.001); and cFn (OR, 1.3; CI95%, 1.1-1.6, p = 0.002) were independently associated with a decrease of neuroserpin levels <70 ng/mL at 24 h after adjusting for confounding factors. CONCLUSIONS: These findings suggest that neuroprotective properties of neuroserpin may be related to the inhibition of excitotoxicity, inflammation, as well as blood brain barrier disruption that occur after acute ischemic stroke

Development of a prototype Lateral Flow Immunoassay (LFI) for the rapid diagnosis of melioidosis

Burkholderia pseudomallei is a soil-dwelling bacterium and the causative agent of melioidosis. Isolation of B. pseudomallei from clinical samples is the &ldquo;gold standard&rdquo; for the diagnosis of melioidosis; results can take 3&ndash;7 days to produce. Alternatively, antibody-based tests have low specificity due to a high percentage of seropositive individuals in endemic areas. There is a clear need to develop a rapid point-of-care antigen detection assay for the diagnosis of melioidosis. Previously, we employed In vivo Microbial Antigen Discovery (InMAD) to identify potential B. pseudomallei diagnostic biomarkers. The B. pseudomallei capsular polysaccharide (CPS) and numerous protein antigens were identified as potential candidates. Here, we describe the development of a diagnostic immunoassay based on the detection of CPS. Following production of a CPS-specific monoclonal antibody (mAb), an antigen-capture immunoassay was developed to determine the concentration of CPS within a panel of melioidosis patient serum and urine samples. The same mAb was used to produce a prototype Active Melioidosis Detect Lateral Flow Immunoassay (AMD LFI); the limit of detection of the LFI for CPS is comparable to the antigen-capture immunoassay (~0.2 ng/ml). The analytical reactivity (inclusivity) of the AMD LFI was 98.7% (76/77) when tested against a large panel of B. pseudomallei isolates. Analytical specificity (cross-reactivity) testing determined that 97.2% of B. pseudomallei near neighbor species (35/36) were not reactive. The non-reactive B. pseudomallei strain and the reactive near neighbor strain can be explained through genetic sequence analysis. Importantly, we show the AMD LFI is capable of detecting CPS in a variety of patient samples. The LFI is currently being evaluated in Thailand and Australia; the focus is to optimize and validate testing procedures on melioidosis patient samples prior to initiation of a large, multisite pre-clinical evaluation