Structure and function in rhodopsin: high level expression of a synthetic bovine opsin gene and its mutants in stable mammalian cell lines.

Stable mammalian cell lines harboring a synthetic bovine opsin gene have been derived from the suspension-adapted HEK293 cell line. The opsin gene is under the control of the immediate-early cytomegalovirus promoter/enhancer in an expression vector that also contains a selectable marker (Neo) governed by a relatively weak promoter. The cell lines expressing the opsin gene at high levels are selected by growth in the presence of high concentrations of the antibiotic geneticin. Under the conditions used for cell growth in suspension, opsin is produced at saturated culture levels of more than 2 mg/liter. After reconstitution with 11-cis-retinal, rhodopsin is purified to homogeneity in a single step by immunoaffinity column chromatography. Rhodopsin thus prepared (> 90% recovery at concentrations of up to 15 microM) is indistinguishable from rhodopsin purified from bovine rod outer segments by the following criteria: (i) UV/Vis absorption spectra in the dark and after photobleaching and the rate of metarhodopsin II decay, (ii) initial rates of transducin activation, and (iii) the rate of phosphorylation by rhodopsin kinase. Although mammalian cell opsin migrates slower than rod outer segment opsin on SDS/polyacrylamide gels, presumably due to a different N-glycosylation pattern, their mobilities after deglycosylation are identical. This method has enabled the preparation of several site-specific mutants of bovine opsin in comparable amounts.

In vivo examination of membrane protein localization and degradation with green fluorescent protein.

To test the utility of green fluorescent protein (GFP) as an in vivo reporter protein when fused to a membrane domain, we made a fusion protein between yeast hydroxymethylglutaryl-CoA reductase and GFP. Fusion proteins displayed spatial localization and regulated degradation consistent with the native hydroxymethylglutaryl-CoA reductase proteins. Thus, GFP should be useful in the study of both membrane protein localization and protein degradation in vivo.

High-level production of recombinant human lysosomal acid alpha-glucosidase in Chinese hamster ovary cells which targets to heart muscle and corrects glycogen accumulation in fibroblasts from patients with Pompe disease.

Infantile Pompe disease is a fatal genetic muscle disorder caused by a deficiency of acid alpha-glucosidase, a glycogen-degrading lysosomal enzyme. We constructed a plasmid containing a 5'-shortened human acid alpha-glucosidase cDNA driven by the cytomegalovirus promoter, as well as the aminoglycoside phosphotransferase and dihydrofolate reductase genes. Following transfection in dihydrofolate reductase-deficient Chinese hamster ovary cells, selection with Geneticin, and amplification with methotrexate, a cell line producing high levels of the alpha-glucosidase was established. In 48 hr, the cells cultured in Iscove's medium with 5 mM butyrate secreted 110-kDa precursor enzyme that accumulated to 91 micrograms.ml-1 in the medium (activity, > 22.6 mumol.hr-1.ml-1). This enzyme has a pH optimum similar to that of the mature form, but a lower Vmax and Km for 4-methylumbelliferyl-alpha-D-glucoside. It is efficiently taken up by fibroblasts from Pompe patients, restoring normal levels of acid alpha-glucosidase and glycogen. The uptake is blocked by mannose 6-phosphate. Following intravenous injection, high enzyme levels are seen in heart and liver. An efficient production system now exists for recombinant human acid alpha-glucosidase targeted to heart and capable of correcting fibroblasts from patients with Pompe disease.

Everything in di Dark Muss Come to Light: A Postcolonial Investigation of the Practice of Extra Lessons at the Secondary Level in Jamaica's Education System

Despite the vast research examining the evolution of Caribbean education systems, little is chronologically tied to the postcolonial theoretical perspectives of specific island-state systems, such as the Jamaican education system and its relationship with the underground shadow education system. This dissertation study sought to address the gaps in the literature by critically positioning postcolonial theories in education to examine the macro- and micro-level impacts of extra lessons on secondary education in Jamaica. The following postcolonial theoretical (PCT) tenets in education were contextualized from a review of the literature: (a) PCT in education uses colonial discourse analysis to critically deconstruct and decolonize imperialistic and colonial representations of knowledge throughout history; (b) PCT in education uses an anti-colonial discursive framework to re-position indigenous knowledge in schools, colleges, and universities to challenge hegemonic knowledge; (c) PCT in education involves the "unlearning" of dominant, normative ideologies, the use of self-reflexivity, and deconstruction; and (d) PCT in education calls for critical pedagogical approaches that reject the banking concept of education and introduces inclusive pedagogy to facilitate "the passage from naïve to critical transitivity" (Freire, 1973, p. 32). Specifically, using a transformative mixed-methods design, grounded and informed by a postcolonial theoretical lens, I quantitatively uncovered and then qualitatively highlighted how if at all extra lessons can improve educational outcomes for students at the secondary level in Jamaica. Accordingly, the quantitative data was used to test the hypotheses that the practice of extra lessons in schools is related to student academic achievement and the practice of critical-inclusive pedagogy in extra lessons is related to academic achievement. The two-level hierarchical linear model analysis revealed that hours spent in extra lessons, average household monthly income, and critical-inclusive pedagogical tents were the best predictors for academic achievement. Alternatively, the holistic multi-case study explored how extra-lessons produces increased academic achievement. The data revealed new ways of knowledge construction and critical pedagogical approaches to galvanize systemic change in secondary education. Furthermore, the data showed that extra lessons can improve educational outcomes for students at the secondary level if the conditions for learning are met. This study sets the stage for new forms of knowledge construction and implications for policy change.

Psychosocial Distress Screening in a Community Cancer Center: An Examination of Gender Differences

Objective: The aim of this study was to gain a better understanding of the needs of male and female oncology patients within a community cancer setting to inform the provision of psychosocial services. Data obtained from 835 single-page measures of oncology patient distress were collected and analyzed to examine the relationship between gender and reported level of distress, the source of this distress, and requests for follow-up from psychosocial service providers.Method: Patients in medical and radiation oncology were given a distress screener tool that included a distress thermometer, a problem checklist, and a list of psychosocial service providers with whom the patient could request to speak.Results: Women reported higher levels of distress than men (p=.003). Women were also more likely than men to endorse practical problems as the cause of their distress (p=.003). A marginally significant relationship between gender and requesting the cancer resource navigator was also found (p=. 059)Conclusion: Gender is a salient factor in reported distress among cancer patients. Although no single variable can entirely explain an individual's response to cancer, male and female patients do appear to have distinctive, gender-specific needs. Psychosocial interventions that account for differences related to gender-role may be particularly beneficial. These results also illustrate the utility of consistent screening practices to better understand and meet the psychosocial needs of oncology

A Cross-Cultural Examination of the Use of Multimodal Metaphor in English and Arabic Cartoons of the ‘Arab Spring’ in Egypt

El estudio de Lakoff y Johnson Metáforas de la vida cotidiana (1980) ha dotado al campo de la metáfora de un renovado interés, y a éste siguieron numerosas publicaciones abordándolo desde diversas perspectivas. Lakoff y Johnson (1980) aseguran que la metáfora es, de hecho, una característica del pensamiento y, por consiguiente, ya no es una propiedad exclusiva del lenguaje. La metáfora sería en realidad un reflejo de cómo concebimos e interpretamos el mundo en que vivimos, en la medida en que está conformada por nuestras experiencias corporales, un fenómeno al que Lakoff y Johnson se refieren como la “mente corpórea”. La metáfora cumple la función de “comprender” y facilitar la interpretación de un ámbito o experiencia en los términos de otro ámbito o experiencia (Lakoff & Johnson, 1980: 36). Los conceptos abstractos son difíciles de ser pensados per se; consecuentemente, las personas recurren a la metáfora para concebirlos en los términos de otras nociones más concretas que están, en cierto grado, vinculadas a nuestras experiencias somáticas. Si bien no escasean las investigaciones en el campo de las metáforas, parece haber, sin embargo, cierto vacío en lo tocante a la manifestación de la metáfora en modos distintos del modo verbal. Forceville (2009: 19) afirma que uno de los principios fundamentales de la Teoría de la Metáfora Conceptual es la idea de que el variado empleo de metáforas por parte del ser humano sugiere “que piensa en gran medida metafóricamente”. Esto lleva a la conclusión de que la manifestación de la metáfora debería estar presente en los diversos modos del pensamiento, y no únicamente en el verbal. Estos otros modos de pensamiento incluyen, entre otros, las imágenes, la música, los sonidos y los gestos. Asimismo, la mayoría de estudios que se han realizado en el área de las metáforas multimodales ha sido aplicada al terreno de la publicidad. Los anunciantes parecen advertir el ! 2! poder que se invierte en las metáforas, y las usan profusamente para transmitir mensajes a los consumidores. No obstante, recientemente existe un nuevo interés por investigar el uso de metáforas multimodales en las caricaturas (véase, por ejemplo, El Refaie 2003, 2009; Schilperoord y Maes 2009; Yus 2009; Bergen 2003; Marin Aresse 2008). Las tiras cómicas se distinguen de los anuncios principalmente por el hecho de que las caricaturas transmiten una postura negativa frente a un sujeto particular, mientras que los anuncios presentan una actitud positiva. Igualmente, les diferencia el hecho de que las tiras cómicas requieren un conocimiento social y político específico. El presente escrito examina y compara el uso de metáforas multimodales en caricaturas políticas – tanto inglesas como árabes – que retratan la Primavera Árabe en Egipto, con el fin de determinar: (1) si la mayoría de caricaturas son conceptualmente específicas o culturalmente específicas; (2) los principales dominios fuente empleados por los caricaturistas ingleses y árabes para interpretar la Primavera Árabe; (3) si hay semejanzas o diferencias entre las caricaturas inglesas y las árabes en su elección del dominio meta; y (4) observar también cómo los distintos modos (aquí esencialmente los modos verbal y pictórico) contribuyen a representar la Primavera Árabe. El corpus para este estudio se compone de un total de 50 tiras cómicas, 25 de ellas inglesas y las 25 restantes árabes. En cada una de estas tiras debe haber al menos una metáfora que representa la Primavera Árabe o alguno de sus subtemas. Las tiras han sido seleccionadas aleatoriamente a través de Internet. Para el análisis, la investigadora ha seguido el marco teórico propuesto por Bounegru y Forceville (2011) en cuanto a la determinación de metáforas en general, así como la determinación de metáforas multimodales de tipo verbovisual en particular. Por cada tira cómica se ha llevado a cabo un análisis independiente, determinando el dominio fuente y el dominio meta, las metáforas, las asignaciones, qué se está representando verbalmente y qué se representa pictóricamente. Además, la autora ha analizado ! 3! posteriormente si estas metáforas son culturalmente específicas o no, y/o si están relacionadas con metáforas más genéricas o universales. El análisis que comprende este trabajo se divide en dos secciones. La primera parte es un estudio detallado de los dos corpora, abordando los distintos escenarios de los dominios fuente empleados por los dibujantes según su similitud. Esta sección presenta a su vez un análisis de los diversos modos que se utilizan para revelar el dominio fuente así como el dominio meta. Del mismo modo, se incluyen las metáforas específicas manejadas en cada tira cómica y, cuando se da el caso, aquellas metáforas genéricas o universales a las que remite. La segunda sección del análisis presenta una comparativa entre ambos corpora basada en el análisis expuesto en la primera parte. Además, trata varios de los fenómenos lingüísticos a los que han recurrido frecuentemente los dibujantes ingleses y árabes. Estos fenómenos son principalmente la metonimia y la personificación. Igualmente, en esta sección la autora investiga en mayor profundidad las metáforas usadas por los dibujantes ingleses y los árabes, determinando si son ora conceptualmente específicas, ora culturalmente específicas, a partir del hecho de que las metáforas sirven como herramienta para reconocer la forma en que personas distintas, o bien culturas distintas, interpretan varias cuestiones. Por ejemplo, a la hora de conceptualizar el futuro, algunas culturas lo representan espacialmente como delante del hablante, mientras que otras se refieren al mismo como localizado detrás del sujeto (Lakoff y Johnson 1980: 14). Este trabajo ha permitido varios hallazgos. En cuanto a las metáforas empleadas para representar la Primavera Árabe, tanto las tiras inglesas como las árabes han recurrido a una cierta variedad de metáforas. Algunas de las tiras presentan más de una metáfora operando a la vez. Los dibujantes ingleses y árabes parecen haberse apoyado siempre en el modo pictórico para presentar el dominio fuente, así como en el modo verbal para mostrar el dominio meta. ! 4! Además, respecto a la naturaleza de las metáforas que figuran en sendos corpora, casi todas las metáforas son conceptuales en cuanto que se corresponden con nuestras experiencias corporales; no hay, por otro lado, ninguna metáfora culturalmente específica. Asimismo, la única diferencia a este respecto entre ambos corpora es una variación en lo que constituye el prototipo de una categoría particular – aquí concretamente la categoría “primavera” en cada una de las culturas –. En las tiras inglesas, una flor es empleada para representar la primavera, mientras que en las árabes el elemento natural más frecuente para simbolizar la primavera es un árbol.

Human Resources (HR) in the Age of Technology: An Examination of the Relationship Between HR and Technology

Technology helps the Human Resources (HR) department drive for strategic relevance. These two departments are successfully collaborating on major projects in such business-critical areas as e-recruiting, self-service, training, compensation and talent management. Technology is critical in helping increase efficiency, increase attraction and retention, reduce administration and cut costs. In recent years, HR information systems (HRIS) have become more important than ever, this time as an essential part of a company's information security and knowledge fields. Ill-suited benefits and disorganized resources are history; now is the time for customized, dynamic plans and connected systems. Employees will appreciate the HRIS, business will benefit from the HRIS and the HR department will no longer have to be the ugly duckling of the company.

Industry Professionals Who Attain Leadership in Energy and Environmental Design (LEED) Accreditation: An Examination of Motivations in Human Resource (HR) Training and Development

Despite the economy, the green building industry continues to grow and drive the demand for environmentally conscious, highly skilled professionals (USGBC 2009). LEED Accredited Professionals (APs) have the knowledge and skills to meet such demand; however, information is limited regarding LEED APs or their motivations and expectations toward prospective employers. The author surveyed a sample of LEED Accredited architects and found a combination of job and personal factors motivated them to attain accreditation. LEED APs value both a competitive salary and commitment to sustainability in prospective employers. To attract, retain, and utilize LEED APs, executives in this industry must reexamine corporate culture, their willingness to pay for credentialing, and the alignment of their reputation with the desires of potential applicants.

Relationship between piezometric level and ground deformations measured by means of DInSAR in the Vega Media of the Segura River (Spain)

Differential SAR Interferometry (DInSAR) is a remote sensing method with the well demonstrated ability to monitor geological hazards like earthquakes, landslides and subsidence. Among all these hazards, subsidence involves the settlement of the ground surface affecting wide areas. Frequently, subsidence is induced by overexploitation of aquifers and constitutes a common problem that affects developed societies. The excessive pumping of underground water decreases the piezometric level in the subsoil and, as a consequence, increases the effective stresses with depth causing a consolidation of the soil column. This consolidation originates a settlement of ground surface that must be withstood by civil structures built on these areas. In this paper we make use of an advanced DInSAR approach - the Coherent Pixels Technique (CPT) [1] - to monitor subsidence induced by aquifer overexploitation in the Vega Media of the Segura River (SE Spain) from 1993 to the present. 28 ERS-1/2 scenes covering a time interval of about 10 years were used to study this phenomenon. The deformation map retrieved with CPT technique shows settlements of up to 80 mm at some points of the studied zone. These values agree with data obtained by means of borehole extensometers, but not with the distribution of damaged buildings, well points and basements, because the occurrence of damages also depends on the structural quality of the buildings and their foundations. The most interesting relationship observed is the one existing between piezometric changes, settlement evolution and local geology. Three main patterns of ground surface and piezometric level behaviour have been distinguished for the study zone during this period: 1) areas where deformation occurs while ground conditions remain altered (recent deformable sediments), 2) areas with no deformation (old and non-deformable materials), and 3) areas where ground deformation mimics piezometric level changes (expansive soils). The temporal relationship between deformation patterns and soil characteristics has been analysed in this work, showing a delay between them. Moreover, this technique has allowed the measurement of ground subsidence for a period (1993-1995) where no instrument information was available.

Season-Long Examination of the Intervention Tone of Coach-Athlete Interactions and Athlete Development in Youth Sport

Objectives: Coaches are a primary influence on athletes' development in youth sport (Horn, 2008). However, the intervention tone of coaches' behaviour has not been directly observed. The purpose of this study was to examine associations between the intervention tone exhibited by youth sport coaches and athletes' individual developmental trajectories over the course of a season. Design: Short-term longitudinal study with behavioural observation. Method: Fifty-five athletes and their coaches from five youth volleyball teams were observed at three time points, and the intervention tone of interactive behaviour was systematically coded and organized by coach-athlete dyad. Athletes completed measures of the 4C's of athlete development (competence, confidence, connection, character) at each time point, which were used to create individualized developmental trajectories. Person-centred analyses were used to examine associations between athletes' developmental trajectories and their unique interactive experiences with their coach. Results: Cluster analysis revealed the presence of three distinct clusters based on athletes' developmental trajectories: 1) high and increasing, 2) low and decreasing, and 3) moderate and maintaining, with athletes from each team distributed across clusters. Analysis of dyadic interaction profiles revealed significant differences in interactive behaviour between clusters. Conclusions: Results suggest that differences in coach-athlete interactive experiences are associated with different developmental trajectories over the course of a season, even for athletes working with the same coach, highlighting the individualized nature of coaches' influence on young athletes. Practical implications for coaches include a critical awareness of their unique interactive relationship with each athlete independently, as well as the importance of fostering these relationships with regard to young people as more than just athletes.

Adolescent Illicit Drug Use and Subsequent Academic and Psychosocial Adjustment : an Examination of Socially-Mediated Pathways

Background: Questions remain regarding the consequences of illicit drug use on adolescent adjustment and the nature of mechanisms that may explain these consequences. In this study, we examined whether early-onset illicit drug use predicts subsequent academic and psychosocial adjustment and whether associations are socially-mediated by decreased school engagement and increased peer deviancy. Method: 4885 adolescents were followed throughout secondary school. We used regressions to determine whether illicit drug use in grade 7 predicted academic achievement, school dropout, depressive symptoms, and conduct problems in grades 10–11, adjusting for potential confounders. We used path analysis to test whether significant associations were mediated by school engagement and peer deviancy in grade 8. Results: Illicit drug use predicted conduct problems and school dropout, but not academic achievement and depressive symptoms. The association between illicit drug use and conduct problems was fully mediated by increased peer deviancy. The association between illicit drug use and school dropout was partially mediated by increased peer deviancy, but remained mostly direct. No indirect association via decreased school engagement was found. Examination of reverse pathways revealed that conduct problems and academic achievement in grade 7 predicted drug use in grades 10–11. These associations were mediated by peer deviancy and school engagement (conduct problems only). Conclusion: Adolescent illicit drug use influences the risk of school dropout and conduct problems in part by contributing to deviant peer affiliation. Reciprocal social mediation characterizes the association between drug use and conduct problems. A reverse mechanism best explains the association with academic achievement.