Time window for recanalization in basilar artery occlusion: Speculative synthesis.

Basilar artery occlusion (BAO) is one of the most devastating forms of stroke and few patients have good outcomes without recanalization. Most centers apply recanalization therapies for BAO up to 12-24 hours after symptom onset, which is a substantially longer time window than the 4.5 hours used in anterior circulation stroke. In this speculative synthesis, we discuss recent advances in BAO treatment in order to understand why and under which circumstances longer symptom duration might not necrotize the brainstem and turn therapeutic attempts futile. We raise the possibility that distinct features of the posterior circulation, e.g., highly developed, persistent collateral arterial network, reverse filling of the distal basilar artery, and delicate plasma flow siding the clot, might sustain brittle patency of brainstem perforators in the face of stepwise growth of the thrombus. Meanwhile, the tissue clock characterizing the rapid necrosis of a typical anterior circulation penumbra will not start. During this perilous time period, recanalization at any point would salvage the brainstem from eventual necrosis caused by imminent reinforcement and further building up of the clot.

Platelet-rich concentrates differentially release growth factors and induce cell migration in vitro

BACKGROUND Platelet-rich concentrates are used as a source of growth factors to improve the healing process. The diverse preparation protocols and the gaps in knowledge of their biological properties complicate the interpretation of clinical results. QUESTIONS/PURPOSES In this study we aimed to (1) analyze the concentration and kinetics of growth factors released from leukocyte- and platelet-rich fibrin (L-PRF), leukocyte- and platelet-rich plasma (L-PRP), and natural blood clot during in vitro culture; (2) investigate the migration of mesenchymal stem cells (MSCs) and human umbilical vein endothelial cells (HUVECs) as a functional response to the factors released; and (3) uncover correlations between individual growth factors with the initial platelet/leukocyte counts or the induced cell migration. METHODS L-PRF, L-PRP, and natural blood clot prepared from 11 donors were cultured in vitro for 28 days and media supernatants collected after 8 hours and 1, 3, 7, 14, and 28 days. Released transforming growth factor β1 (TGF-β1), vascular endothelial growth factor (VEGF), insulin growth factor (IGF-1), platelet-derived growth factor AB (PDGF-AB), and interleukin-1β (IL-1β) were measured in the supernatants with enzyme-linked immunosorbent assay. Migration of MSC and HUVEC induced by the supernatants was evaluated in Boyden chambers. RESULTS More TGF-ß1 was released (mean ± SD in pg/mL of blood) from L-PRF (37,796 ± 5492) compared with L-PRP (23,738 ± 6848; p < 0.001) and blood clot (3739 ± 4690; p < 0.001), whereas more VEGF and IL-1ß were released from blood clot (1933 ± 704 and 2053 ± 908, respectively) compared with both L-PRP (642 ± 208; p < 0.001 and 273 ± 386; p < 0.001, respectively) and L-PRF (852 ± 376; p < 0.001 and 65 ± 56, p < 0.001, respectively). No differences were observed in IGF-1 and PDGF-AB released from any of the concentrates. TGF-β1 release peaked at Day 7 in L-PRF and at 8 hours and Day 7 in L-PRP and 8 hours and Day 14 in blood clot. In all concentrates, main release of VEGF occurred between 3 and 7 days and of IL-1β between Days 1 and 7. IGF-1 and PDGF-AB were released until Day 1 in L-PRP and blood clot, in contrast to sustained release over the first 3 days in L-PRF. The strongest migration of MSC occurred in response to L-PRF, and more HUVEC migration was seen in L-PRF and blood clot compared with L-PRP. TGF-β1 correlated with initial platelet counts in L-PRF (Pearson r = 0.66, p = 0.0273) and initial leukocyte counts in L-PRP (Pearson r = 0.83, p = 0.0016). A positive correlation of IL-1β on migration of MSC and HUVEC was revealed (Pearson r = 0.16, p = 0.0208; Pearson r = 0.31, p < 0.001). CONCLUSIONS In comparison to L-PRP, L-PRF had higher amounts of released TGF-β1, a long-term release of growth factors, and stronger induction of cell migration. Future preclinical studies should confirm these data in a defined injury model. CLINICAL RELEVANCE By characterizing the biologic properties of different platelet concentrates in vitro, we may gain a better understanding of their clinical effects and develop guidelines for specific future applications.

Three-year changes of prothrombotic factors in a cohort of South Africans with a high clinical suspicion of obstructive sleep apnea.

A hypercoagulable state might be one important mechanism linking obstructive sleep apnea (OSA) with incident myocardial infarction and stroke. However, previous studies on prothrombotic factors in OSA are not uniform and cross-sectional. We longitudinally studied prothrombotic factors in relation to OSA risk, adjusting for baseline levels of prothrombotic factors, demographics, metabolic parameters, aspirin use, and life style factors. The Berlin Questionnaire and/or neck circumference were used to define high OSA risk in 329 South African teachers (48.0 % male, 44.6 % black) at baseline and at three-year follow-up. Von Willebrand factor (VWF), fibrinogen, D-dimer, plasminogen activator inhibitor-1, clot lysis time (CLT), and soluble urokinase-type plasminogen activator receptor (suPAR) were measured in plasma. At baseline 35.7 % of participants had a high risk of OSA. At follow-up, persistently high OSA risk, persistently low OSA risk, OSA risk remission, and new-onset OSA risk were present in 26.1 %, 53.2 %, 9.4 %, and 11.3 % of participants, respectively. New-onset OSA risk was associated with a significant and longitudinal increase in VWF, fibrinogen, CLT, and suPAR relative to persistently low OSA risk; in VWF, fibrinogen, and suPAR relative to remitted OSA risk; and in VWF relative to persistently high OSA risk. Persistently high OSA risk was associated with an increase in CLT and suPAR relative to persistently low OSA risk and in D-dimer relative to remitted OSA risk. Remitted OSA risk was associated with D-dimer decrease relative to persistently low OSA risk. In OSA, hypercoagulability is a dynamic process with a most prominent three-year increase in individuals with new-onset OSA risk.

MASP-1 Induced Clotting--The First Model of Prothrombin Activation by MASP-1.

Mannan-binding lectin-associated serine protease-1 (MASP-1), a protein of the complement lectin pathway, resembles thrombin in terms of structural features and substrate specificity. Due to its interplay with several coagulation factors, it has the ability to induce fibrin clot formation independent of the usual coagulation activation pathways. We have recently shown that MASP-1 activates prothrombin and identified arginine (R) 155, R271, and R393 as potential cleavage sites. FXa cleaves R320 instead of R393, and thrombin cleaves R155 and R284 in prothrombin. Here we have used three arginine-to-glutamine mutants of prothrombin, R271Q, R320Q, R393Q and the serine-to-alanine active site mutant S525A to investigate in detail the mechanism of MASP-1 mediated prothrombin activation. Prothrombin wildtype and mutants were digested with MASP-1 and the cleavage products were analysed by SDS-PAGE and N-terminal sequencing. A functional clotting assay was performed by thrombelastography. We have found that MASP-1 activates prothrombin via two simultaneous pathways, either cleaving at R271 or R393 first. Both pathways result in the formation of several active alternative thrombin species. Functional studies confirmed that both R393 and R320 are required for prothrombin activation by MASP-1, whereas R155 is not considered to be an important cleavage site in this process. In conclusion, we have described for the first time a detailed model of prothrombin activation by MASP-1.

Studies of the structure and function of a fibrinogen binding MSCRAMM(RTM) from Staphylococcus epidermidis

Coagulase-negative staphylococci (CNS) are recognized as important pathogens and are particularly associated with foreign body infections. S. epidermidis accounts for approximately 75% of the infections caused by CNS. Three genes, sdrF, sdrG, and sdrH, were identified by screening a S. epidermidis genomic library with a probe encompassing the serine-aspartate dipeptide repeat-encoding region (region R) of clfA from S. aureus. SdrG has significant amino acid identity to ClfA, ClfB and other surface proteins of S. aureus. SdrG is also similar to a protein (Fbe) recently described by Nilsson, et al. (Infection and Immunity, 1998, 66:2666–73) from S. epidermidis. The N-terminal domain (A region) of SdrG was expressed as a his-tag fusion protein in E. coli. In an ELISA, this protein, rSdrG(50-597) was shown to bind specifically to fibrinogen (Fg). Western ligand blot analysis showed that SdrG binds the Bβ chain of Fg. To further characterize the rSdrG(50-597)-Fg interaction, truncates of the Fg Bβ chain were made and expressed as recombinant proteins in E. coli. SdrG was shown to bind the full-length Bβ chain (1462), as well as the N-terminal three-quarters (1-341), the N-terminal one-half (1-220) and the N-terminal one-quarter (1-95) Bβ chain constructs. rSdrG(50-597) failed to bind to the recombinant truncates that lacked the N-terminal 25 amino acid residues of this polypeptide suggesting that SdrG recognizes a site within this region of the Bβ chain. Inhibition ELISAs have shown that peptide mimetics, including β1–25, and β6–20, encompassing this 25 residue region can inhibit binding of rSdrG(50-597) to Fg coated wells. Using fluorescence polarization we were able to determine an equilibrium constant (KD) for the interaction of rSdrG(50-597) with the Fg Bβ chain peptide β1–25. The labeled peptide was shown to bind to rSdrG(50-597) with a KD of 0.14 ± 0.01μM. Because rSdrG(50-597) recognizes a site in the Fg Bβ chain close to the thrombin cleavage site, we investigated the possibility of the rSdrG(50-597) site either overlapping or lying close to this cleavage site. An ELISA showed that rSdrG(50-597) binding to thrombin-treated Fg was significantly reduced. In a clot inhibition assay rSdrG(50-597) was able to inhibit fibrin clot formation in a concentration dependent manner. Furthermore, rSdrG(50-597) was able to inhibit clot formation by preventing the release of fibrinopeptide B as determined by HPLC. To further define the interaction between rSdrG(50-597) and peptide β6–20, we utilized an alanine amino acid replacement strategy. The residues in β6–20 that appear to be important in rSdrG(50-597) binding to Fg, were confirmed by the rSdrG(273-597)-β6–20 co-crystal structure that was recently solved by our collaborators at University of Alabama-Birmingham. Additionally, rSdrG(50-597) was not able to bind to Fg from different animal species, rather it bound specifically to human Fg in an ELISA. This suggests that the sequence variation between Fg Bβ chains of different species, specifically with in the N-terminal 25 residues, affects the ability of rSdrG(50-597) binding to Fg, and this may explain why S. epidermidis is primarily a human pathogen. ^

Interactions and structural analysis of the zinc -binding motif in decorin, a small leucine rich proteoglycan in extracellular matrix

Decorin, a dermatan/chondroitin sulfate proteoglycan, is ubiquitously distributed in the extracellular matrix (ECM) of mammals. Decorin belongs to the small leucine rich proteoglycan (SLRP) family, a proteoglycan family characterized by a core protein dominated by Leucine Rich Repeat motifs. The decorin core protein appears to mediate the binding of decorin to ECM molecules, such as collagens and fibronectin. It is believed that the interactions of decorin with these ECM molecules contribute to the regulation of ECM assembly, cell adhesions, and cell proliferation. These basic biological processes play critical roles during embryonic development and wound healing and are altered in pathological conditions such as fibrosis and tumorgenesis. ^ In this dissertation, we discover that decorin core protein can bind to Zn2+ ions with high affinity. Zinc is an essential trace element in mammals. Zn2+ ions play a catalytic role in the activation of many enzymes and a structural role in the stabilization of protein conformation. By examining purified recombinant decorin and its core protein fragments for Zn2+ binding activity using Zn2+-chelating column chromatography and Zn2+-equilibrium dialysis approaches, we have located the Zn2+ binding domain to the N-terminal sequence of the decorin core protein. The decorin N-terminal domain appears to contain two Zn2+ binding sites with similar high binding affinity. The sequence of the decorin N-terminal domain does not resemble any other reported zinc-binding motifs and, therefore, represents a novel Zn 2+ binding motif. By investigating the influence of Zn2+ ions on decorin binding interactions, we found a novel Zn2+ dependent interaction with fibrinogen, the major plasma protein in blood clots. Furthermore, a recombinant peptide (MD4) consisting of a 41 amino acid sequence of mouse decorin N-terminal domain can prolong thrombin induced fibrinogen/fibrin clot formation. This suggests that in the presence of Zn2+ the decorin N-terminal domain has an anticoagulation activity. The changed Zn2+-binding activities of the truncated MD4 peptides and site-directed mutagenesis generated mutant peptides revealed that the functional MD4 peptide might contain both a structural zinc-binding site in the cysteine cluster region and a catalytic zinc site that could be created by the flanking sequences of the cysteine cluster region. A model of a loop-like structure for MD4 peptide is proposed. ^

Dispositivos de formación profesional : Diálogos entre el campo de la investigación y de la formación profesional

En esta comunicación se intentan identificar algunas de las características específicas que pueden presentar los dispositivos de formación profesional de futuros docentes. A partir de los resultados de una investigación sobre la construcción de saberes pedagógicos en la acción desarrollada en el seno de un Grupo Profesional de Referencia en la que se recurre a autoconfrontaciones simples y cruzadas (Clot, 2004), se analizan los aportes teórico-metodológicos para la puesta en acción de dispositivos de formación profesional durante la formación inicial. La orientación teórico-metodológica adoptada pretende con la intención de construir saberes pedagógicos desde una perspectiva profesional contribuir a la generación de situaciones de formación. Dichas situaciones serían potencialmente capaces de hacer emerger, movilizar y poner en acción los saberes pedagógicos construidos por los formadores y por los estudiantes. Se propone entonces diseñar situaciones de formación que recuperen distintas culturas de acción pedagógica, construyendo nuevos sentidos en interacción con el medio profesional, el corpus disciplinar de la Pedagogía y la propia situación de formación. Finalmente se intenta avanzar sobre dos grupos de interrogantes a saber: La acción entendida como conjunto de actividades dotadas de una unidad de sentido y/o de significaciones en el ámbito de interacciones con otros sujetos (Barbier and Galatanu, 2000) se presentaría casi "invisible" a los ojos de los educadores en los dispositivos de formación más tradicionales ¿Cómo descubrir la acción en las situaciones de formación? Los dispositivos de formación profesional centrados en la emergencia de saberes pedagógicos en la acción trascienden la lógica secuencial de pensamiento y acción ¿Cómo poner en acción, cómo movilizar el saber pedagógico del formador en situaciones de formación? ¿Cuáles son los aportes que puede ofrecer el análisis de la actividad (Clot, 2004)? Es intención de esta ponencia proponer una serie de cuestiones que resitúen los aportes de la Pedagogía en el campo de la Formación contribuyendo a la formulación de nuevas preguntas que permitan orientar la acción.

Dispositivos de formación profesional : Diálogos entre el campo de la investigación y de la formación profesional

En esta comunicación se intentan identificar algunas de las características específicas que pueden presentar los dispositivos de formación profesional de futuros docentes. A partir de los resultados de una investigación sobre la construcción de saberes pedagógicos en la acción desarrollada en el seno de un Grupo Profesional de Referencia en la que se recurre a autoconfrontaciones simples y cruzadas (Clot, 2004), se analizan los aportes teórico-metodológicos para la puesta en acción de dispositivos de formación profesional durante la formación inicial. La orientación teórico-metodológica adoptada pretende con la intención de construir saberes pedagógicos desde una perspectiva profesional contribuir a la generación de situaciones de formación. Dichas situaciones serían potencialmente capaces de hacer emerger, movilizar y poner en acción los saberes pedagógicos construidos por los formadores y por los estudiantes. Se propone entonces diseñar situaciones de formación que recuperen distintas culturas de acción pedagógica, construyendo nuevos sentidos en interacción con el medio profesional, el corpus disciplinar de la Pedagogía y la propia situación de formación. Finalmente se intenta avanzar sobre dos grupos de interrogantes a saber: La acción entendida como conjunto de actividades dotadas de una unidad de sentido y/o de significaciones en el ámbito de interacciones con otros sujetos (Barbier and Galatanu, 2000) se presentaría casi "invisible" a los ojos de los educadores en los dispositivos de formación más tradicionales ¿Cómo descubrir la acción en las situaciones de formación? Los dispositivos de formación profesional centrados en la emergencia de saberes pedagógicos en la acción trascienden la lógica secuencial de pensamiento y acción ¿Cómo poner en acción, cómo movilizar el saber pedagógico del formador en situaciones de formación? ¿Cuáles son los aportes que puede ofrecer el análisis de la actividad (Clot, 2004)? Es intención de esta ponencia proponer una serie de cuestiones que resitúen los aportes de la Pedagogía en el campo de la Formación contribuyendo a la formulación de nuevas preguntas que permitan orientar la acción.

Using the Spectrum Analyzer as an Educational Tool for Mobile Communications

Application of the spectrum analyzer for illustrating several concepts associated with mobile communications is discussed. Specifically, two groups of observable features are described. First, time variation and frequency selectivity of multipath propagation can be revealed by carrying out simple measurements on commercial-network GSM and UMTS signals. Second, the main time-domain and frequency-domain features of GSM and UMTS radio signals can be observed. This constitutes a valuable tool for teaching mobile communication courses.

An investigation of the performance of a new Mechanical thrombectomy device using Bond Graph modelling: application to the extraction of blood clots in the middle cerebral artery

A number of thrombectomy devices using a variety of methods have now been developed to facilitate clot removal. We present research involving one such experimental device recently developed in the UK, called a ‘GP’ Thrombus Aspiration Device (GPTAD). This device has the potential to bring about the extraction of a thrombus. Although the device is at a relatively early stage of development, the results look encouraging. In this work, we present an analysis and modeling of the GPTAD by means of the bond graph technique; it seems to be a highly effective method of simulating the device under a variety of conditions. Such modeling is useful in optimizing the GPTAD and predicting the result of clot extraction. The aim of this simulation model is to obtain the minimum pressure necessary to extract the clot and to verify that both the pressure and the time required to complete the clot extraction are realistic for use in clinical situations, and are consistent with any experimentally obtained data. We therefore consider aspects of rheology and mechanics in our modeling.

Regulación, Participación y Agricultura Urbana. Análisis Legislativo, Normativo y de Modelos de Gestión en Londres, Berlín y Madrid

En general las ciudades europeas cuentan con una amplia tradición de agricultura urbana, en la que las asociaciones de hortelanos juegan un papel importante en la gestión de los espacios y en la interlocución con las administraciones públicas. Por su parte los gobiernos locales desarrollan políticas de fomento de los huertos urbanos y han definido herramientas concretas para su regulación, protección y gestión. Esta doble tradición ha sido decisiva a la hora de resistir las presiones urbanizadoras que se producen en el suelo urbano. En el estado español no encontramos el mismo contexto normativo y las iniciativas de agricultura urbana están aún en periodo de ensayo, aunque son numerosos los programas municipales de huertos de ocio y están aumentando los proyectos liderados por asociaciones vecinales y ecologistas. Basándonos en los resultados del análisis se propone un modelo administrativo para el desarrollo de la agricultura urbana en las ciudades españolas, que parta de la definición de políticas concretas, el reconocimiento normativo en los planes de ordenación urbanística y en las ordenanzas municipales, y el diseño de un modelo de gestión en el que participe el gobierno local (adquisición de terrenos, infraestructuras, asistencia técnica y financiera, espacio de coordinación) y las asociaciones de hortelanos. Palabras clave: normativa urbanística, participación ciudadana, políticas públicas, huertos comunitarios.

Historia de los huertos urbanos. De los huertos para pobres a los programas de agricultura urbana ecológica

Para entender el papel actual de la agricultura urbana (AU) es necesaria una revisión histórica de sus funciones y características en la ciudad occidental, estudiando los motivos de su implantación, su sentido, evolución y potencialidades. Los momentos de mayor auge de la AU están ligados a crisis económicas y energéticas, que obligan a recurrir a ella para asegurar el autoabastecimiento. En los inicios de la ciudad industrial del XIX la AU cumple funciones de subsistencia, higiene y control social. En países como Gran Bretaña, Alemania o Francia las autoridades locales y las grandes fábricas se ven obligadas a ofrecer terrenos a los trabajadores para completar sus recursos y mejorar las condiciones de vida en los barrios obreros. En la primera mitad del siglo XX la AU aparece ligada a las guerras mundiales, su función es de subsistencia y patriótica, de apoyo a la economía de guerra y a los procesos de posguerra. Se desarrollan programas gubernamentales y campañas de fomento de la AU, como Dig for Victory en Gran Bretaña, o Victory Gardens en Estados Unidos. A partir de los años 70 los proyectos de AU cumplen funciones de desarrollo local, integración social y educación ambiental, y son lideradas por organizaciones comunitarias y ecologistas. En el momento actual la AU tiene la potencialidad de ser un instrumento de mejora ambiental y social, colaborando en la sostenibilidad urbana, la lucha contra el cambio climático, la calidad de vida y la creación de ciudades a escala humana. Palabras clave: rehabilitación urbana, autonomía alimentaria, metabolismo urbano, calidad de vida.

Applicability of the GP Device to the Circle of Willis Arteries by Using a Mathematical Model

According to the World Health Organization, 15 million people suffer stroke worldwide each year, of these, 5 million die and 5 million are permanently disabled. Stroke is therefore a major cause of mortality world-wide. The majority of strokes are caused by a blood clot that occludes an artery in the brain, and although thrombolytic agents such as Alteplase are used to dissolve clots that arise in the arteries of the brain, there are limitations on the use of these thrombolytic agents. However over the past decade, other methods of treatment have been developed which include Thrombectomy Devices e.g. the 'GP' Thrombus Aspiration Device ('GP' TAD). Such devices may be used as an alternative to thrombolytics or in conjunction with them to extract blood clots in arteries such as the middle cerebral artery of the midbrain brain, and the posterior inferior cerebellar artery (PICA) of the posterior aspect of the brain. In this paper, we mathematically model the removal of blood clots using the 'GP' TAD from selected arteries of the brain where blood clots may arise taking into account factors such as the resistances, compliances and inertances effects. Such mathematical modelling may have potential uses in predicting the pressures necessary to extract blood clots of given lengths, and masses from arteries in the Circle of Willis - posterior circulation of the brain

Analysis and design of a thrombectomy device by using simulation techniques

Purpose: In this work, we present the analysis, design and optimization of one experimental device recently developed in the UK, called the 'GP' Thrombus Aspiration Device (GPTAD). This device has been designed to remove blood clots without the need to make contact with the clot itself thereby potentially reducing the risk of problems such as downstream embolisation. Method: To obtain the minimum pressure necessary to extract the clot and to optimize the device, we have simulated the performance of the GPTAD analysing the resistances, compliances and inertances effects. We model a range of diameters for the GPTAD considering different forces of adhesion of the blood clot to the artery wall, and different lengths of blood clot. In each case we determine the optimum pressure required to extract the blood clot from the artery using the GPTAD, which is attached at its proximal end to a suction pump. Result: We then compare the results of our mathematical modelling to measurements made in laboratory using plastic tube models of arteries of comparable diameter. We use abattoir porcine blood clots that are extracted using the GPTAD. The suction pressures required for such clot extraction in the plastic tube models compare favourably with those predicted by the mathematical modelling. Discussion & Conclusion: We conclude therefore that the mathematical modelling is a useful technique in predicting the performance of the GPTAD and may potentially be used in optimising the design of the device.

Simulation of a mechanical thrombectomy device based in the use of self-expandable stents for the blood clots extraction

Recently, we have presented some studies concerning the analysis, design and optimization of one experimental device developed in the UK - GPTAD - which has been designed to remove blood clots without the need to make contact with the clot itself, thereby potentially reducing the risk of problems such as downstream embolisation. Based on the idea of a modification of the previous device, in this work, we present a model based in the use of stents like the SolitaireTM FR, which is in contact with the clot itself. In the case of such devices, the stent is self-expandable and the extraction of the blood clot is faciliatated by the stent, which must be inside the clot. Such stents are generally inserted in position by using the guidewire inserted into the catheter. This type of modeling could potentially be useful in showing how the blood clot is moved by the various different forces involved. The modelling has been undertaken by analyzing the resistances, compliances and inertances effects. We model an artery and blood clot for range of forces for the guidewire. In each case we determine the interaction between blood clot, stent and artery.