The development of plasmatic glutamate grabbers for the treatment of ischemic stroke / Desarrollo de atrapadores de glutamato plasmático como terapia para el ictus isquémico

Glutamate is subjected to strong homeostasis, which is regulated by excitatory amino acid transporters (EAATs), being the EAAT2 responsible for 90% of the uptake of extracellular glutamate. The balance disturbance of glutamate, as in ischemic stroke occurs, provokes that glutamate increases in the extracellular space which induces a complex process of pathogenic mechanisms. In order to mitigate these effects, neuroprotective drugs were developed, which showed no efficacy in human clinical practice. Because of this, studies describe the existence of glutamate concentration gradient between the endothelial cells that form the blood-brain barrier and the bloodstream. When the glutamate concentration of the brain endothelial cells is greater than the concentration of blood glutamate, the glutamate is carried from the brain to the bloodstream. On account of, the hypothesis that reducing blood levels of glutamate would increase the concentration gradient between brain and blood endothelium arises, thus favoring the removal of cerebral glutamate. This potential neuroprotective mechanism is called blood glutamate grabbering. For all the above, the combination of mechanisms of neuroprotection (by blood glutamate grabbing) and neurorepair (mediated by mesenchymal stem cells) constitute a promising therapeutic strategy in cerebral ischemia. Therefore, the objective of this work was the development of alternative blood glutamate grabbers for use in ischemic stroke. So we have used two strategies: enzymatic and cellular. The aim of enzymatic strategy was to analyze the protective effect of the human rGOT1 alone and in combination with a non-effective dose of oxaloacetate in an animal model of ischemic stroke. Sixty rats were subjected to a transient middle cerebral artery occlusion (MCAO). Infarct volumes were assessed by magnetic resonance imaging (MRI) before treatment administration, and 24 h and 7 days after MCAO. Brain glutamate levels were determined by in vivo MR spectroscopy (MRS) during artery occlusion (80 min) and reperfusion (180 min). GOT activity and serum glutamate concentration were analyzed during the occlusion and reperfusion period. Somatosensory test was performed at baseline and 7 days after MCAO. The three treatments (6.44 μg/100g, 12.88 μg/100g and 25.76 μg/100g) tested induced a reduction in serum and brain glutamate levels, resulting in a reduction in infarct volume and sensorimotor deficit; being the dose of 12.88 μg/100 g the most effective. Protective effect of rGOT1 supplemented with oxaloacetate at 7 days persists even when treatment was delayed until at least 2 h after onset of ischemia. In conclusion, our findings indicate that the combination of human rGOT1 with low doses of oxaloacetate seems to be a successful approach for stroke treatment

Towards the Recycling of Bio-Waste: The Case of Pontevedra, Spain (REVITALIZA)

Waste management is one of the main environmental problems that municipalities have to address. The fulfilment of the recycling objectives imposed by the European Community requires the segregation and treatment of the municipal bio-waste. Pontevedra Provincial Council started in 2015 an innovative plan, called REVITALIZA, for the recycling of bio-waste through the promotion of composting in municipalities. REVITALIZA, which is developed in different phases, advocates the implementation of local composting (home and community composting) and small composting facilities, so that the generation of waste and the economic and environmental costs of its collection and transport are reduced. The plan is a pioneer in the training of technical personnel in the area of bio-waste management. Currently, 36 municipalities are participating in REVITALIZA in different phases of the plan, committed to locally managing bio-waste

Higher expression of Toll-like receptors 2 and 4 in blood cells of Keratoconus patiens

Inflammation may play a significant role in Keratoconus (KC), but the implication of immunity on this inflammatory response is unknown. Therefore, our aim was to determine the expression levels of Toll-like receptors 2 (TLR2) and 4 (TLR4) in monocytes and neutrophils from patients with KC and control subjects for demonstrating the role of innate immunity in KC. We also study the correlation between TLR2/TLR4 expression and serum levels of proinflammatory markers (IL-1β, IL-6, TNF-α, MMP-9 and NF-κB). Forty patients with bilateral KC (55% males; mean age; 33.1 ± 10.9 years) and 20 control subjects (55% males; mean age; 30.4 ± 7.6 years) were included in the study. Our results showed that mean expression of TLR2 and TLR4 in both neutrophils and monocytes was significantly higher in patients with KC compared to control subjects (all p < 0.0001). Furthermore, KC patients also showed higher serum levels of IL-1B, IL-6, TNF-α, MMP-9 (all p < 0.0001) and NF-κB (p = 0.036). In addition, we found a strong correlation between TLR2 expression in both monocytes and neutrophils (all p < 0.0001), and TLR4 in monocytes (all p < 0.05) with serum levels of IL-1B, IL-6, TNF-α and MMP-9. In conclusion, these findings suggest that TLRs may play an important role in the pathophysiology of KC.This project has been supported by grants from the Spanish Ministry of Economy and Competitiveness (Instituto de Salud Carlos III: PI14/00247); Xunta de Galicia (Consellería de Economía e Industria: 10 PX113918156PR and Consellería de Educación: GRC2014/027) and the European Union program FEDER. T. Sobrino (CP12/03121) and F. Campos (CP14/00154) are recipients of a research contract from Miguel Servet Program of Instituto de Salud Carlos III. Likewise A. Vieites-Prado is recipient of a FPI fellow (BES-2012-056027) from the Spanish Ministry of Economy and Competitiveness; and U. Regueiro is recipient of a predoctoral Fellow from the Xunta de Galicia (ED481A-2015/001)S

Hyperthermia in human ischemic and hemorrhagic stroke: similar outcome, different mechanisms

Hyperthermia is a predictor of poor outcome in ischemic (IS) and intracerebral hemorrhagic (ICH) stroke. Our aim was to study the plausible mechanisms involved in the poor outcome associated to hyperthermia in stroke. We conducted a case-control study including patients with IS (n = 100) and ICH (n = 100) within the first 12 hours from symptom onset. Specifically, IS and ICH patients were consecutively included into 2 subgroups, according to the highest body temperature within the first 24 hours: Tmax 2) at 3 months. Serum levels of glutamate and active MMP-9 were measured at admission. Our results showed that Tmax ≥37.5°C within the first 24 hours was independently associated with poor outcome in both IS (OR, 12.43; 95% CI, 3.73-41.48; p<0.0001) and ICH (OR, 4.29; 95% CI, 1.32-13.91; p = 0.015) after adjusting for variables with a proven biological relevance for outcome. However, when molecular markers levels were included in the logistic regression model, we observed that glutamate (OR, 1.01; 95% CI, 1.00-1.02; p = 0.001) and infarct volume (OR, 1.06; 95% CI, 1.01-1.10; p = 0.015) were the only variables independently associated to poor outcome in IS, and active MMP-9 (OR, 1.04; 95% CI, 1.00-1.08; p = 0.002) and National Institute of Health Stroke Scale (NIHSS) at admission (OR, 1.29; 95% CI, 1.13-1.49; p<0.0001) in ICH. In conclusion, these results suggest that although the outcome associated to hyperthermia is similar in human IS and ICH, the underlying mechanisms may be different

Human recombinant glutamate oxaloacetate transaminase 1 (GOT1) supplemented with oxaloacetate induces a protective effect after cerebral ischemia

Blood glutamate scavenging is a novel and attractive protecting strategy to reduce the excitotoxic effect of extracellular glutamate released during ischemic brain injury. Glutamate oxaloacetate transaminase 1 (GOT1) activation by means of oxaloacetate administration has been used to reduce the glutamate concentration in the blood. However, the protective effect of the administration of the recombinant GOT1 (rGOT1) enzyme has not been yet addressed in cerebral ischemia. The aim of this study was to analyze the protective effect of an effective dose of oxaloacetate and the human rGOT1 alone and in combination with a non-effective dose of oxaloacetate in an animal model of ischemic stroke. Sixty rats were subjected to a transient middle cerebral artery occlusion (MCAO). Infarct volumes were assessed by magnetic resonance imaging (MRI) before treatment administration, and 24 h and 7 days after MCAO. Brain glutamate levels were determined by in vivo MR spectroscopy (MRS) during artery occlusion (80 min) and reperfusion (180 min). GOT activity and serum glutamate concentration were analyzed during the occlusion and reperfusion period. Somatosensory test was performed at baseline and 7 days after MCAO. The three treatments tested induced a reduction in serum and brain glutamate levels, resulting in a reduction in infarct volume and sensorimotor deficit. Protective effect of rGOT1 supplemented with oxaloacetate at 7 days persists even when treatment was delayed until at least 2 h after onset of ischemia. In conclusion, our findings indicate that the combination of human rGOT1 with low doses of oxaloacetate seems to be a successful approach for stroke treatment.Ministeiro de Economía y Competitividad de EspañaXunta de Galicia /Consellería Economía IndustriaXunta de Galicia/ Consellería EducaciónInstituto de Salud Carlos IIISpanish Research Network on Cerebrovascular Diseases RETICS-INVICTUSFundación Mútua MadrileñaEuropean Union program FEDEREspaña. Ministerio de Economía y Competitividad/SAF2011-30517Xunta de Galicia /Consellería Economía Industria/10PXIB918282PRXunta de Galicia / Consellería Educación/ CN2011/010Instituto de Salud Carlos III/PI11/00909Instituto de Salud Carlos III/CP12/03121Spanish Research Network on Cerebrovascular Diseases RETICS-INVICTUS /RD12/0014Instituto de Salud Carlos III/PI10/00449Instituto de Salud Carlos III/PI12/0311

Recognition of AMP, ADP and ATP through cooperative binding by Cu(II) and Zn(II) complexes containing urea and/or phenylboronic acid moieties

[Abstract] We report a series of Cu(II) and Zn(II) complexes with different ligands containing a dipicolyl unit functionalized with urea groups that may contain or not a phenylboronic acid function. These complexes were designed for the recognition of phosphorylated anions through coordination to themetal ion reinforced by hydrogen bonds involving the anion and NHgroups of urea. The complexes were isolated and several adducts with pyrophosphate were characterized using X-ray diffraction measurements. Coordination of one of the urea nitrogen atoms to themetal ion promoted the hydrolysis of the ligands containing 1,3-diphenylurea units, while ligands bearing 1-ethyl-3-phenylurea groups did not hydrolyze significantly at room temperature. Spectrophotometric titrations, combined with 1H and 31P NMR studies, were used in investigating the binding of phosphate, pyrophosphate (PPi), and nucleoside 50-polyphosphates (AMP, ADP, ATP, CMP, and UMP). The association constants determined in aqueous solution (pH 7.0, 0.1MMOPS) point to a stronger association with PPi, ADP, and ATP as compared with the anions containing a single phosphate unit. The [CuL4]2+ complex shows important selectivity for pyrophosphate (PPi) over ADP and ATP.Galicia. Consellería de Cultura, Educación e Ordenación Universitaria; EM 2012/08

Increased Endothelial Progenitor Cell Levels are Associated with Good Outcome in Intracerebral Hemorrhage

Circulating endothelial progenitor cells (EPCs) play a role in the regeneration of damaged brain tissue. However, the relationship between circulating EPC levels and functional recovery in intracerebral hemorrhage (ICH) has not yet been tested. Therefore, our aim was to study the influence of circulating EPCs on the outcome of ICH. Forty-six patients with primary ICH (males, 71.7%; age, 72.7 ± 10.8 years) were prospectively included in the study within 12 hours of symptom onset. The main outcome variable was good functional outcome at 12 months (modified Rankin scale ≤2), considering residual volume at 6 months as a secondary variable. Circulating EPC (CD34+/CD133+/KDR+) levels were measured by flow cytometry from blood samples obtained at admission, 72 hours and day 7. Our results indicate that patients with good outcome show higher EPC numbers at 72 hours and day 7 (all p < 0.001). However, only EPC levels at day 7 were independently associated with good functional outcome at 12 months (OR, 1.15; CI95%, 1.01–1.35) after adjustment by age, baseline stroke severity and ICH volume. Moreover, EPC levels at day 7 were negatively correlated to residual volume (r = −0.525; p = 0.005). In conclusion, these findings suggest that EPCs may play a role in the functional recovery of ICH patients.This study has been partially supported by grants from Instituto de Salud Carlos III (PI14/01879), Spanish Research Network on Cerebrovascular Diseases RETICS-INVICTUS (RD12/0014), Xunta de Galicia (Consellería Educación GRC2014/027) and the European Union program FEDER. Furthermore, F. Campos (CP14/00154) and T. Sobrino (CP12/03121) are recipients of a research contract from Miguel Servet Program of Instituto de Salud Carlos IIIS

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

Supercritical water oxidation for energy production by hydrothermal flame as internal heat source. Experimental results and energetic study

Producción CientíficaThe cooled wall reactor has been modified by adding an additional upper outlet of products at 500–700 °C to improve energy recovery and make possible energy generation with the supercritical water oxidation of different waste. Experimental and modeling results of the performance of this new reactor configuration are presented as well as a theoretical analysis of the energy recovery of the reactor compared to other supercritical water oxidation reactors. Different flow distributions were tested to find the best elimination conditions. Total organic carbon removal over 99.99% was obtained at room injection temperatures, when the fraction of products leaving the reactor in the upper effluent is lower than 70% of feed flow. The performance of the reactor was tested with the oxidation of a recalcitrant compound such as ammonia. Removals higher than 99% of were achieved at temperatures near 700 °C. The behavior of the reactor working with feeds with up to 2.5% wt Na2SO4 could be injected in the reactor without plugging problems. Upper effluent always presented a concentration of salt lower than 30 ppm. Theoretical energetic analysis shows that the performance of this reactor is superior to other designs obtaining a maximum power efficiency of 27% (0.339 kW/kg-feed).Ministerio de Economía, Industria y Competitividad - FEDER (Proyect CTQ2013-44143-R

Mordida cruzada anterior y tratamiento en la atención primaria

Introducción: la mordida cruzada anterior es la descripción de una anomalía de la oclusión en el plano antero-posterior donde los dientes inferiores están delante de los superiores. Objetivo: evaluar la efectividad del plano inclinado en la corrección de  las Mordidas Cruzadas Anteriores Simples (MCAS). Material y Método: se diseñó un estudio analítico, transversal y aplicado en  escolares matriculados desde preescolar a sexto grado en escuelas primarias del área de salud del Policlínico “Pedro Borrás Astorga”, Municipio Pinar del Río 2013-2014. El universo estuvo constituido por 55 niños afectados por Mordidas Cruzadas Anteriores (MCA) de ambos sexos y con edades entre 4-11 años mientras la  muestra se conformó por 28 niños afectados por MCAS a los que se le confeccionaron aparatos de plano inclinado de acrílico sobre modelos de yeso, los pacientes fueron citados semanalmente para control y activación. Los resultados se calcularon a través del test de chi cuadrado con nivel de significación de p < 0,05. Resultados: se definió que 9.2% de los escolares estaban afectados por mordidas cruzadas anteriores, correspondiendo el 51 % a las Mordidas Cruzadas Anteriores Simples, el sexo masculino fue el más afectado y el grupo de edad de 7-9 años. El 96% de esta anomalía fue corregida en un tiempo no mayor de 21 días. Conclusiones: las MCA son un problema de salud que enfrenta la población infantil. Se recomienda el plano inclinado como un método sencillo, rápido, eficaz y bien tolerado por los pacientes al alcance del Estomatólogo General Integral