Heparin and cancer revisited: Mechanistic connections involving platelets, P-selectin, carcinoma mucins, and tumor metastasis

Independent studies indicate that expression of sialylated fucosylated mucins by human carcinomas portends a poor prognosis because of enhanced metastatic spread of tumor cells, that carcinoma metastasis in mice is facilitated by formation of tumor cell complexes with blood platelets, and that metastasis can be attenuated by a background of P-selectin deficiency or by treatment with heparin. The effects of heparin are not primarily due to its anticoagulant action. Other explanations have been suggested but not proven. Here, we bring together all these unexplained and seemingly disparate observations, showing that heparin treatment attenuates tumor metastasis in mice by inhibiting P-selectin-mediated interactions of platelets with carcinoma cell-surface mucin ligands. Selective removal of tumor mucin P-selectin ligands, a single heparin dose, or a background of P-selectin deficiency each reduces tumor cell-platelet interactions in vitro and in vivo. Although each of these maneuvers reduced the in vivo interactions for only a few hours, all markedly reduce long-term organ colonization by tumor cells. Three-dimensional reconstructions by using volume-rendering software show that each situation interferes with formation of the platelet “cloak” around tumor cells while permitting an increased interaction of monocytes (macrophage precursors) with the malignant cells. Finally, we show that human P-selectin is even more sensitive to heparin than mouse P-selectin, giving significant inhibition at concentrations that are in the clinically acceptable range. We suggest that heparin therapy for metastasis prevention in humans be revisited, with these mechanistic paradigms in mind.

Mechanistic coupling of protease signaling and initiation of coagulation by tissue factor

The crucial role of cell signaling in hemostasis is clearly established by the action of the downstream coagulation protease thrombin that cleaves platelet-expressed G-protein-coupled protease activated receptors (PARs). Certain PARs are cleaved by the upstream coagulation proteases factor Xa (Xa) and the tissue factor (TF)–factor VIIa (VIIa) complex, but these enzymes are required at high nonphysiological concentrations and show limited recognition specificity for the scissile bond of target PARs. However, defining a physiological mechanism of PAR activation by upstream proteases is highly relevant because of the potent anti-inflammatory in vivo effects of inhibitors of the TF initiation complex. Activation of substrate factor X (X) by the TF–VIIa complex is here shown to produce enhanced cell signaling in comparison to the TF–VIIa complex alone, free Xa, or Xa that is generated in situ by the intrinsic activation complex. Macromolecular assembly of X into a ternary complex of TF–VIIa–X is required for proteolytic conversion to Xa, and product Xa remains transiently associated in a TF–VIIa–Xa complex. By trapping this complex with a unique inhibitor that preserves Xa activity, we directly show that Xa in this ternary complex efficiently activates PAR-1 and -2. These experiments support the concept that proinflammatory upstream coagulation protease signaling is mechanistically coupled and thus an integrated part of the TF–VIIa-initiated coagulation pathway, rather than a late event during excessive activation of coagulation and systemic generation of proteolytic activity.

Ras-catalyzed hydrolysis of GTP: a new perspective from model studies.

Despite the biological and medical importance of signal transduction via Ras proteins and despite considerable kinetic and structural studies of wild-type and mutant Ras proteins, the mechanism of Ras-catalyzed GTP hydrolysis remains controversial. We take a different approach to this problem: the uncatalyzed hydrolysis of GTP is analyzed, and the understanding derived is applied to the Ras-catalyzed reaction. Evaluation of previous mechanistic proposals from this chemical perspective suggests that proton abstraction from the attacking water by a general base and stabilization of charge development on the gamma-phosphoryl oxygen atoms would not be catalytic. Rather, this analysis focuses attention on the GDP leaving group, including the beta-gamma bridge oxygen of GTP, the atom that undergoes the largest change in charge in going from the ground state to the transition state. This leads to a new catalytic proposal in which a hydrogen bond from the backbone amide of Gly-13 to this bridge oxygen is strengthened in the transition state relative to the ground state, within an active site that provides a template complementary to the transition state. Strengthened transition state interactions of the active site lysine, Lys-16, with the beta-nonbridging phosphoryl oxygens and a network of interactions that positions the nucleophilic water molecule and gamma-phosphoryl group with respect to one another may also contribute to catalysis. It is speculated that a significant fraction of the GAP-activated GTPase activity of Ras arises from an additional interaction of the beta-gamma bridge oxygen with an Arg side chain that is provided in trans by GAP. The conclusions for Ras and related G proteins are expected to apply more widely to other enzymes that catalyze phosphoryl (-PO(3)2-) transfer, including kinases and phosphatases.

Mechanistic Importance of Redox Potentials and Conformational Flexibility in Electron-Transferring Flavoproteins

The mitochondrial matrix flavoproteins electron transfer flavoprotein (ETF) and electron transfer flavoprotein-ubiquinone oxidoreductase (ETF-QO) are responsible for linking fatty acid β-oxidation with the main mitochondrial respiratory chain. Electrons derived from flavoprotein dehydrogenases are transferred sequentially through ETF and ETF-QO to ubiquinone and then into the respiratory chain via complex III. In this study, the effects of changes in ETF-QO redox potentials on its activity and the conformational flexibility of ETF were investigated. ETF-QO contains one [4Fe-4S]2+,1+ and one flavin adenine dinucleotide (FAD). In the porcine protein, threonine 367 is hydrogen bonded to N1 and O2 of the flavin ring of the FAD. The analogous site in Rhodobacter sphaeroides ETF-QO is asparagine 338. Mutations N338T and N338A were introduced into the R. sphaeroides protein by site-directed mutagenesis to determine the impact of hydrogen bonding at this site on redox potentials and activity. FAD redox potentials were measured by potentiometric titration probed by electron paramagnetic resonance (EPR) spectroscopy. The N338T and N338A mutations lowered the midpoint potentials, which resulted in a decrease in the quinone reductase activity and negligible impact on disproportionation of ETF1e- catalyzed by ETF-QO. These observations indicate that the FAD is involved in electron transfer to ubiquinone, but not in electron transfer from ETF to ETF-QO. Therefore it is proposed that the iron-sulfur cluster is the immediate acceptor from ETF. It has been proposed that the αII domain of ETF is mobile, allowing promiscuous interactions with structurally different partners. Double electron-electron resonance (DEER) was used to measure the distance between spin labels at various sites and an enzymatically reduced FAD cofactor in Paracoccus denitrificans ETF. Two or three interspin distance distributions were observed for spin-labels in the αI (A43C) and βIII (A111C) domains, but only one is observed for a label in the βII (A210C) domain. This suggests that the αII domain adopts several stable conformations which may correspond to a closed/inactive conformation and an open/active conformation. An additional mutation, E162A, was introduced to increase the mobility of the αII domain. The E162A mutation doubled the activity compared to wild-type and caused the distance distributions to become wider. The DEER method has the potential to characterize conformational changes in ETF that occur when it interacts with various redox partners.

Utilizing Evidence-Based Psychology Practice in the University of Denver's Professional Psychology Center: A Proposal

As acceptance of the Evidence-based Psychology Practice (EBPP) model continues to grow (Pagoto, Spring, Coups, Mulvaney, Coutu, & Ozakinci, 2007), it seems pertinent to explore how this model can be applied in different settings. This topic is timely as practitioners in the field are being held ever more accountable for the efficacy of the treatments they employ (Pagoto et al., 2007). Increased scrutiny has resulted in a need to integrate research into practice in order to ensure continued relevance in the ever-changing realm of American health care (Luebbe, Radcliffe, Callands, Green & Thorn, 2007; Collins, Leffingwell & Belar, 2007; Chwalisz, 2003). This paper explores how the requirements set forth by the American Psychological Association Presidential Task Force on Evidence-Based Practice (2006) can be implemented at the University of Denver's (DU) Professional Psychology Center (PPC), a training clinic for students enrolled in the Psy.D. program at DU's Graduate School of Professional Psychology (GSPP). In doing so, the methods employed by Collins et al. (2007) at Oklahoma State University (OSU) are used as a template and modified to accommodate differences between these two institutions.

Photogenerated o-Azaxylylenes: Mechanistic Studies and Synthetic Applications

This research sets out to build upon excited state o-azaxylylene cycloaddition. The mechanism behind the excitation and cycloaddition process of photogenerated o-azaxylylenes was determined experimentally. Time-correlated single-photon counting, steady-state spectroscopy, triplet quenching experiments, and quantum yield studies provided evidence suggesting that excited state intramolecular proton transfer is followed by intersystem crossing and stepwise addition to the tethered unsaturated pendant. In keeping with the principles of diversity oriented synthesis, a modular approach was taken to gain access to a diverse array of N,O,S-Polyheterocycles which were modified postphotochemically via Suzuki coupling to yield fused biaryls. Cycloaddition products, outfitted with halogens in the aromatic ring of the o-azaxylylene, proved to be reactive with a variety of boronic acids resulting in a rapid growth in structural complexity. A novel procedure was developed that utilized multiple o-azaxylylene cores in a photochemical cascade transformation yielding complex scaffolds of unprecedented topology. The photoprecursors were produced in a one-pot two-step sequence from commercially available starting materials, and upon irradiation yield structures containing up to five fused hetrocyclic rings, and showed complete diastereoselectivity.

A proposal of algorithm for solving definite descriptions through dialogue structure

En este trabajo proponemos un algoritmo para la resolución de las descripciones definidas en español a través de la estructura del diálogo, mediante la definición de un espacio de accesibilidad anafórico. Este algoritmo está basado en la hipótesis de que la resolución de la anáfora está relacionada con la estructura del diálogo. Así, la resolución de la anáfora mejora si se especifica un espacio de accesibilidad para cada tipo descripción definida según la estructura del diálogo. La utilización de este espacio de accesibilidad anafóico reduce tanto el tiempo de procesamiento como la posibilidad de obtener un antecedente erróneo. Además, la definición de este espacio de accesibilidad depende únicamente de la propia estructura textual del diálogo.

A general proposal to multilingual information access based on syntactic-semantic patterns

In the last few years, there has been a wide development in the research on textual information systems. The goal is to improve these systems in order to allow an easy localization, treatment and access to the information stored in digital format (Digital Databases, Documental Databases, and so on). There are lots of applications focused on information access (for example, Web-search systems like Google or Altavista). However, these applications have problems when they must access to cross-language information, or when they need to show information in a language different from the one of the query. This paper explores the use of syntactic-sematic patterns as a method to access to multilingual information, and revise, in the case of Information Retrieval, where it is possible and useful to employ patterns when it comes to the multilingual and interactive aspects. On the one hand, the multilingual aspects that are going to be studied are the ones related to the access to documents in different languages from the one of the query, as well as the automatic translation of the document, i.e. a machine translation system based on patterns. On the other hand, this paper is going to go deep into the interactive aspects related to the reformulation of a query based on the syntactic-semantic pattern of the request.

Assessing participation in online discussion forums: a proposal for multidimensional analysis

This paper aims to assess the discussion forums being used in the programme Especialista Universitario online sobre Tecnologías de la Información y la Comunicación, taught at the University of Alicante, on the basis of the analysis model suggested by Kay (2004). In such a model, it is essential to represent graphically the forum activity so that the visual representation may improve analysis. This research has allowed reaching results which define the forum activity and has contributed with a proposal for multi analysis in the area of assessing participation in communication within online discussion forums.

Perspectivas sobre educación en valores en tiempos de crisis

En medio de la actual crisis económica mundial, todas las instituciones de la sociedad están afectadas por la crisis moral, de manera que existe una evidente y preocupante contradicción entre ellas. Esta crisis moral implica la carencia de valores personales y ciudadanos, que afectan, significativamente, al desarrollo del individuo y a la convivencia social. Tratamos de verificar si hay indicios de ello en la realidad escolar con relación a la axiología curricular y su desarrollo. Para analizar la situación de la enseñanza-aprendizaje de los valores en la educación española, hemos aplicado una encuesta a 1320 profesores de educación secundaria, con el fin de averiguar los objetivos, los tipos de valores y las habilidades personales y sociales, que desarrollan en el aula. La situación nos indica que son necesarios nuevos enfoques de educación en valores, que se adapten a estos tiempos. La formación axiológica debe estar incorporada en todos los ámbitos del conocimiento y ha de ir a la par que la formación de la inteligencia. El binomio actividad intelectual-actividad moral ha de formar un todo insociable. En esta propuesta, reflexionamos en torno a la aplicación de un nuevo paradigma axiológico que se separe del racionalismo mecanicista, y que se oriente a una educación personal holística, sistemática, interdisciplinaria y transversal a todo el currículo educativo.

Towards a new proposal to evaluate the learning objects quality in learning strategies for education (QEES)

In Computer Science world several proposals have been developed for the assessment of the quality of the digital objects, based on the capabilities and facilities offered by current technologies and the available resources. Years ago researchers and specialists from both educational and technological areas have been committed to the development of strategies that improve the quality of education. At present, in the field of teaching-learning, another important aspect is the need to improve the manner of gaining knowledge and learning in education, which the use of learning strategies is a major advance in the teaching-learning process in institutions of higher education. This paper presents QEES, a proposal for evaluating the quality of the learning objects employed on learning strategies to support students during their education processes by using information extraction techniques and ontologies.

La 'otra ciudad' funcionalista. El caso del Cabo de Las Huertas (1959-1979)

Se analiza la zona de la playa San Juan, el Cabo de las Huertas y la Albufereta, situadas a 5 km al NE de la ciudad de Alicante, durante el periodo (1959-79), en el contexto de la Dictadura y con el único marco legal de la Ley del Suelo española 1956. Cabría decir que, en 1963, aparece la Ley de Centros y Zonas de Interés Turístico Nacional. El cambio de las condiciones nacionales de 1959 propicia la urbanización de la playa de San Juan en el cabo de Huertas de Alicante que, ya formaba parte de una ambiciosa propuesta de 1933. Juan Guardiola Gaya, arquitecto catalán, redacta el plan parcial de esta franja de terrenos frente al mar para convertirla en un barrio turístico autónomo. Resulta significativo que elementos tradicionalmente antiurbanos como la naturaleza, los espacios libres y la arquitectura aislada son los que definan esta nueva ciudad para las vacaciones. De ahí que durante mucho tiempo estos núcleos turísticos vacíos en gran parte del año y constituidos solamente casi por segundas residencias no hayan sido considerados auténticas ciudades. Obviamente estos núcleos, a diferencia de las ciudades propiamente dichas, eran ciudades-satélites alojadas en la periferia litoral y cultural. No disponían de equipamiento suficiente pero tampoco contaban con industrias. Por el contrario, tenían más espacios libres. Utilizamos el término ciudad, a pesar de su tamaño, porque estaban pensadas para el comercio del tiempo libre. Eran las otras ciudades, las del verano para las vacaciones, como este caso.

Proposal of a New Parameter for the Weathering Characterization of Carbonate Flysch-Like Rock Masses: The Potential Degradation Index (PDI)

The susceptibility of clay bearing rocks to weathering (erosion and/or differential degradation) is known to influence the stability of heterogeneous slopes. However, not all of these rocks show the same behaviour, as there are considerable differences in the speed and type of weathering observed. As such, it is very important to establish relationships between behaviour quantified in a laboratory environment with that observed in the field. The slake durability test is the laboratory test most commonly used to evaluate the relationship between slaking behaviour and rock durability. However, it has a number of disadvantages; it does not account for changes in shape and size in fragments retained in the 2 mm sieve, nor does its most commonly used index (Id2) accurately reflect weathering behaviour observed in the field. The main aim of this paper is to propose a simple methodology for characterizing the weathering behaviour of carbonate lithologies that outcrop in heterogeneous rock masses (such as Flysch slopes), for use by practitioners. To this end, the Potential Degradation Index (PDI) is proposed. This is calculated using the fragment size distribution curves taken from material retained in the drum after each cycle of the slake durability test. The number of slaking cycles has also been increased to five. Through laboratory testing of 117 samples of carbonate rocks, extracted from strata in selected slopes, 6 different rock types were established based on their slaking behaviour, and corresponding to the different weathering behaviours observed in the field.

A proposal for teaching with authentic texts in order to develop social awareness in a language subject in tertiary education

The University of the 21st century has to establish links with society and prepare students for the demands of the working world. Therefore, this article is a contribution to the integral preparation of university students by proposing the use of authentic texts with social content in English lessons so that students acquire emotional and social competencies while still learning content. This article will explain how the choice of texts on global issues such as racism and gender helps students to develop skills such as social awareness and critical thinking to deepen their understanding of discrimination, injustice or gender differences in both oral and written activities. A proposal will be presented which involves using the inauguration speech from Mandela's presidency and texts with photographs of women so that students analyse them whilst utilising linguistic tools that allow them to explore a text's social dimension.