Investigation of the roles of CheD in the Bacillus subtilis chemotactic signal transduction

Abstract The two-component based chemotaxis signal transduction system allows flagellated bacteria to sense their surrounding chemical environment and move towards more favorable conditions. The attractant signals can be sensed by transmembrane chemoreceptors, and then transmitted to the histidine kinase CheA. Once activated, CheA interacts with the response regulator CheY through phosphorelay, which causes a change in the rotation of the flagella. The direction of flagella rotation determines whether a cell swims straight or just tumbles. Cells also need adaptation to respond to a change in chemical concentrations, and return to their prestimulated level. Adaptation in the B. subtilis chemotaxis system is achieved by three coordinated systems: the methylation system, the CheC/CheD/CheY-p system and the CheV system. CheD, the previously identified receptor deamidase, was shown to be critical to the ability of B. subtilis to perform chemotaxis and is the main focus of this study. This study started from characterization of the enzymatic mechanism of CheD. Results showed that CheD deamidase uses a cysteine hydrolase mechanism. The catalytic triad consisting of Cys33-His50-Thr27, and Ser27 is essential for receptor recognition and binding. In addition, in this study CheC was found to inhibit CheD’s deamidase activity. Through mutant screening, Phe102 on CheD was found to be the essential site to interact with CheC. Furthermore, the CheD/CheC interaction is necessary for the robust chemotaxis in vivo as demonstrated by the cheD (F102E) mutant, which lacks the ability to swim on swarm plates. Despite its deamidase activity, we hypothesized that CheD’s main role is its involvement in the CheD-CheC-CheY-p negative feedback pathway during adaptation. In particular, CheD is likely to help stabilize the transient kinase-activating state through binding to receptors. When CheY-p level is increased, CheC-CheY-p complex may attract CheD away from receptors. In this study, CheC-CheD binding kinetics with CheY or CheYp presence was successfully obtained by a series of SPR experiments. The increased affinity of CheD for CheC in presence of CheYp but not CheY makes likely the hypothesis that CheC-CheD-CheY interact as part of a negative feedback pathway during adaptation. Last, the interaction between CheD and chemoreceptor McpC was studied in order to better understand the role of CheD in adaptation. Results showed that Q304 and Q305 on McpC are essential to recruit CheD. Additionally, the reduced levels of CheD in mcpC (Q304A) or (Q305A) mutants suggested that the dynamic interaction between CheD and receptors is vital to maintain the normal CheD level. These findings suggest more complicated roles of CheD than its previously identified function as a receptor deamidase, and will lead to a clearer picture of the coordination of the three adaptational systems in the B. subtilis chemotactic sensory transduction system.

Visión y tratamiento educativo de los roles masculino y femenino desde el punto de vista de los videojugadores: tecnologías emergentes favorecedoras de la igualdad de género

Los videojuegos representan uno de los productos de ocio de mayor consumo entre ambos géneros. Los personajes de los videojuegos suelen ser exagerados y generadores de estereo tipos, principalmente el femenino, que no proyectan un personaje creíble, con el que se puedan identificar las jugadoras. Se propone una clasificación de arquetipos para catalogar los personajes femeninos presentes en los videojuegos. Se ha utilizado una metodología exploratoria para indagar los estereotipos existentes en los videojuegos junto con los estereotipos de género. Finalmente, se ha diseñado un programa de actividades para trabajar con videojuegos y género en el aula de modo que esté enfocado para que los alumnos empaticen con personajes de otro género y así puedan derribarse tanto los prejuicios como los estereotipos para trabajar en pro de la igualdad.

The Descent of Pluto: Interactive dynamics, specialisation and reciprocity of roles in a Wikipedia debate

International audience

Working with Student Teachers: A Mixed Methods Study to Examine the Roles and Self-Identified Dispositions of Cooperating Teachers

A critical component of teacher education is the field experience during which candidates practice under the supervision of experienced teachers. Programs use the InTASC Standards to define the requisite knowledge, skills, and dispositions for teaching. Practicing teachers are familiar with the concepts of knowledge and skills, but they are less familiar with dispositions. Practicing teachers who mentor prospective teachers are underrepresented in the literature, but they are critical to teacher preparation. The research goals were to describe the self-identified dispositions of cooperating teachers, identify what cooperating teachers consider their role in preparing prospective teachers, and explain challenges that cooperating teachers face. Using a mixed methods design, I conducted a quantitative survey followed by a qualitative case study. When I compared survey and case study data, cooperating teachers report possessing InTASC critical dispositions described in Standard 2: Learning Differences, Standard 3: Learning Environments, and Standard 9: Professional Learning and Ethical Practice, but not Standard 6: Assessment and Standard 10: Leadership and Collaboration. Cooperating teachers assume the roles of modeler, mentor and advisor, and informal evaluator. They explain student teachers often lack skills and dispositions to assume full teaching responsibilities and recommend that universities better prepare candidates for classrooms. Cooperating teachers felt university evaluations were not relevant to teaching reality. I recommend modifying field experiences to increase the quantity and duration of classroom placements. I suggest further research to detail cooperating teacher dispositions, compare cooperating teachers who work with different universities, and determine if cooperating teacher dispositions influence student teacher dispositions.

Tutoría en E-Learning. Funciones y roles del tutor en la formación online

La teleformación, de manera paralela a Internet, ha tenido una rápida y constante evolución a lo largo de los últimos años. Al mismo tiempo, dicha modalidad formativa ha ido cobrando mucha relevancia en el ámbito de la formación, en particular, en la capacitación de las personas trabajadoras. Teniendo en cuenta las ventajas que presenta la teleformación, se hace necesario que se realicen avances y mejoras de la calidad en el desarrollo de los procesos incluidos en ella, como es el caso de la tutorización. Por estas razones esta investigación se plantea: ¿Qué funciones realizan los tutores y tutoras de e-learning durante las acciones formativas en las que participan?, y ¿cómo perciben esas funciones los estudiantes? Este trabajo investiga las formas de tutorización que ayudan a un mejor desempeño por parte del teletutor, promoviendo procesos de enseñanza-aprendizaje que contribuyan a disminuir el índice de abandonos de estudiantes. La metodología seguida emplea técnicas cuantitativas y cualitativas buscando en cada momento, el método que mejor dé respuesta a los objetivos planteados inicialmente. Para la realización de la misma se han tomado dos muestras. Una de 707 estudiantes de cursos online pertenecientes a un proyecto de formación continua de trabajadores de pequeña y mediana empresas, y autónomos, La segunda muestra la conforman 8 tutores que han participado en el mencionado proyecto. A través de esta investigación se han podido analizar y estudiar las dimensiones y funciones que han realizado los tutores durante las distintas acciones formativas llevadas a cabo y se ha llegado a conocer las percepciones que tienen los estudiantes sobre las distintas dimensiones o funciones que ha empleado cada tutor en las mismas. Se consiguió identificar y comprender las tareas y funciones puestas en práctica por los tutores durante las acciones formativas llevadas a cabo, y las diferencias que se produjeron en ellas en cada uno de los docentes. Así mismo los resultados han permitido organizar diversos sistemas de categorías en otros tantos marcos “teóricos”, representados a través de diagramas comprensivos, que nos ayudan a entender de manera más adecuada las relaciones entre categorías y dimensiones de cada uno de los tutores del estudio. Ha sido posible incluso llegar a establecer tipologías de tutores según las funciones y roles desarrollados, los recursos más utilizados en su labor de tutorización y los rasgos que caracterizan a los estudiantes que realizan este tipo de formación.

TESTING THE JOINT ROLES OF CAREER DECISION SELF-EFFICACY AND PERSONALITY TRAITS IN THE PREDICTION OF CAREER INDECISION

Career decision-making self-efficacy and the Big Five traits of neuroticism, extraversion, and conscientiousness were examined as predictors of career indecision in a sample of 181 undergraduates. Participants completed an online survey. I predicted that the Big Five traits and career decision-making self-efficacy would (a) interrelate moderately and (b) each relate significantly and moderately to career indecision. In addition, I predicted that career decision-making self-efficacy would partially mediate the relationships between the Big Five traits and career indecision, while the Big Five traits were predicted to moderate the relationship between career decision-making self-efficacy and career indecision. Finally, I predicted that career decision-making self-efficacy would account for a greater amount of unique variance in career indecision than the Big Five traits. All predicted correlations were significant. Career decision-making self-efficacy fully mediated the relationship of Extraversion to career indecision and partially mediated the relationships of Neuroticism and Conscientiousness to career indecision. Conscientiousness was found to moderate the relationship of career decision-making self-efficacy to career indecision such that the negative relation between self-efficacy and career indecision was stronger in the presence of high conscientiousness. This study builds upon existing research on the prediction of career indecision by examining potential mediating and moderating relationships.

REDUCING TEACHER ATTRITION: THE ROLES AND RESPONSIBILITIES OF BUILDING PRINCIPALS IN A LARGE URBAN DISTRICT TO SUPPORT TEACHER RETENTION

The high rate of teacher attrition in urban schools is well documented. While this does not seem like a problem in Carter County, this equates to hundreds of teachers that need to be replaced annually. Since school year (SY) 2007-08, Carter County has lost over 7,100 teachers, approximately half of (50.1%) of whom resigned, often going to neighboring, higher-paying jurisdictions as suggested by exit survey data (SY2016-2020 Strategic Plan). Included in this study is a range of practices principals use to retain teachers. While the role of the principal is recognized as a critical element in teacher retention, few studies explore the specific practices principals implement to retain teachers and how they use their time to accomplish this task. Through interviews, observations, document analysis and reflective notes, the study identifies the practices four elementary school principals of high and relatively low attrition schools use to support teacher retention. In doing so, the study uses a qualitative cross-case analysis approach. The researcher examined the following leadership practices of the principal and their impact on teacher retention: (a) providing leadership, (b) supporting new teachers, (c) training and mentoring teaching staff, (d) creating opportunities for collaboration, (d) creating a positive school climate, and (e) promoting teacher autonomy. The following research questions served as a foundational guide for the development and implementation of this study: 1. How do principals prioritize addressing teacher attrition or retention relative to all of their other responsibilities? How do they allocate their time to this challenge? 2. What do principals in schools with low attrition rates do to promote retention that principals in high attrition schools do not? What specific practices or interventions are principals in these two types of schools utilizing to retain teachers? Is there evidence to support their use of the practices? The findings that emerge from the data revealed the various practices principals use to influence and support teachers do not differ between the four schools.

Las redes sociales y sus roles. Algunas experiencias

Desde 2007, el fenómeno más destacado de intemet ha sido el rápido crecimiento de las redes sociales. Una de cada cuatro personas en el mundo las usa actualmente según Team Family Guy (Infographic Journal, dic. 2013). El tiempo mensual promedio dedicado a ellas por visitante alcanzó 23,7 horas en 2013 a nivel global y a 24 horas en América Latina, donde los brasileños son los que les dedican más tiempo (32,9hrs), seguidos de los argentinos (22hrs) y los peruanos ( 19,9hrs) (Datos de FayerWayer, oc. 2013). Facebook ha pasado el billón americano -un millón de millones de páginas vistas en un mes en agosto 2011 y domina en Europa y América. Facebook terminó 2013 con más de 1.150 millones de usuarios mensuales, Google+ con más de 300 millones, Linkedln con 259 millones y Twitter con 240 millones (ABC, oct.2013). Cada día, el 62,8% de los internautas españoles navega por las redes sociales, según el Primer Estudio de Medios de Comunicación Online presentado por IAB Spain (PRnoticias, 19/02/2014).

Unemployment, parental distress and youth emotional well-being: The moderation roles of parent-youth relationship and financial deprivation

We investigated, in a sample of 112 unemployed parents of adolescents aged 10-19 years, the links between parental distress and change in youth emotional problems related to parental unemployment, and the moderation roles of parent-youth relationship and financial deprivation. Data were analyzed using descriptive statistics and correlations. Further, simple moderation, additive moderation, and moderated moderation models of regression were performed to analyze the effects of parental distress, parent-youth relationship and financial deprivation in predicting change in youth emotional problems related to parental unemployment. Results show that parental distress moderated by parent-youth relationship predicted levels of change in youth emotional problems related to parental unemployment. This study provides evidence that during job loss, parental distress is linked to youth emotional well-being and that parent-youth relationships play an important moderation role. This raises the importance of further researching parental distress impacts on youth well-being, especially during periods of high unemployment rates.

Investigation of the functional roles of host cell proteins involved in Coronavirus infection using highly specific and scalable RNA interference (RNAi) approach

Since its identification in the 1990s, the RNA interference (RNAi) pathway has proven extremely useful in elucidating the function of proteins in the context of cells and even whole organisms. In particular, this sequence-specific and powerful loss-of-function approach has greatly simplified the study of the role of host cell factors implicated in the life cycle of viruses. Here, we detail the RNAi method we have developed and used to specifically knock down the expression of ezrin, an actin binding protein that was identified by yeast two-hybrid screening to interact with the Severe Acute Respiratory Syndrome Coronavirus (SARS-CoV) spike (S) protein. This method was used to study the role of ezrin, specifically during the entry stage of SARS-CoV infection.

Dual Roles of Diadenosine Polyphosphates in Corneal Epithelial Cell Migration

Purpose. To investigate the influence of diadenosine polyphosphates on the rate of corneal epithelial cell migration. Methods. Primary corneal epithelial cell cultures were obtained from New Zealand White rabbits. Immunocytochemical experiments were performed by fixing the cells with 4% paraformaldehyde (PFA) and incubated with cytokeratin 3 primary antibody, which was subsequently incubated with a secondary IgG mouse labeled with FITC, and the cells were observed under confocal microscopy. Migration studies were performed by taking confluent monolayers that were wounded with a pipette tip and challenged with different di- and mononucleotides with or without P2 antagonist (n = 8 each treatment). For concentration–response analysis, compounds were tested in doses ranging from 10−8 to 10−3 M (n = 8). The stability of the dinucleotides was assayed by HPLC, with an isocratic method (n = 4). Results. Cells under study were verified as corneal epithelial cells via the immunocytochemical analysis. Cell migration experiments showed that Ap4A, UTP, and ATP accelerated the rate of healing (5, 2.75, and 3 hours, respectively; P < 0.05; P < 0.001), whereas Ap3A, Ap5A, and UDP delayed it (6.5, 10, and 2 hours, respectively; P < 0.05). ADP did not modify the rate of migration. Antagonists demonstrated that Ap4A and Ap3A did activate different P2Y receptors mediating corneal wound-healing acceleration and delay. Concerning the possible degradation of the dinucleotides, it was almost impossible to detect any products resulting from their cleavage. Conclusions. Based on the pharmacological profile of all the compounds tested, the two main P2Y receptors that exist in these corneal cells are a P2Y2 receptor accelerating the rate of healing and a P2Y6 receptor that delays this process.

Pathways to adulthood and their precursors: Roles of gender and race

Research on the transition to adulthood dates back nearly four decades, but a growing body of research has taken a new approach by investigating multiple demographic markers in the transition to adulthood simultaneously. Using the life course perspective, this dissertation is built on the literature by first examining contemporary young adults’ pathways to adulthood from ages 18 to 30 and their differences by gender. Data for this study were drawn from the National Longitudinal Survey of Youth 1997; the final sample included 2,185 men and 2,086 women. The college-educated single workers pathway, the college-educated married working parents pathway, and the high-school-educated single parents pathway were identified in both genders. For men, the study also identified the high-school-educated single workers pathway and the high-school-educated married working parents pathway. For women, the study also identified the high-school-educated workers pathway and the high-school-educated married parents pathway. Not only did the definitions of some pathways differ by gender, but even in the pathways with the same definition, gender differences were found in the probabilities of being married, of being a parent, or of being employed full-time. Based on the pathways to adulthood identified, this research examined the family and adolescent precursors and whether race moderates the associations between family structure experiences and young adults’ pathways to adulthood. Parental education, family structure, GPA, delinquency, early sexual activity, and race/ethnicity were the family and adolescent precursors that distinguished among pathways taken by the youth. Two interactions between race and family structure/instability were identified. The positive association between growing up in a single-parent family and the odds of taking the high-school-educated single workers pathway compared to the college-educated married working parents pathway was weaker for Black males than for White males. The positive association between family instability and the odds of taking the college-educated single workers pathway compared to the college-educated married working parents pathway was weaker for Black females than for White females. This dissertation accounted for changes in the multiple statuses related to becoming an adult by following contemporary young adults for 12 years. More research on contemporary young adults’ pathways to adulthood and subgroup differences in the effects of precursors are recommended. Limitations and implications of this study are discussed.