Surface Mutation Thr34His Facilitates Purification of Haemophilus influenza Carbonic Anhydrase via Metal Affinity Chromatography

In order to pursue Haemophilus influenza carbonic anhydrase (HICA) as a potential drug target, easy and efficient purification methods must be developed. While immobilized metal affinity chromatography (IMAC) may be used, complications with polyhistidine tags is a concern. Inspired by the endogenous metal affinity of Escherichia coli β-carbonic anhydrase (ECCA), we suggest that the generation of histidine clusters on HICA’s surface will facilitate its purification by metal affinity chromatography without the potential interference of His-tags. Here we investigate the Thr34His mutation as a method to generate metal affinity in HICA. Since Thr34His is located only 5.3 Å away from His32, the two residues make a vicinal histidine pair that can interact with nickel resin. We report successful generation of Thr34His HICA mutant plasmid via site-directed mutagenesis. To obtain mutant protein for metal affinity chromatography, Thr34His HICA was overexpressed in E. coli cells and isolated as a cell lysate with a concentration of 20.2 ± 0.6 mg/mL. Metal affinity chromatography was performed on the sample, and the chromatography fractions were analyzed by SDS-PAGE in order to assess the metal affinity of the mutant. SDS-PAGE revealed that while Thr34His HICA eluted at low 10 mM and 25 mM concentrations of imidazole, 150 mM imidazole was required to fully elute the mutant. These results suggest that through the generation of surface histidine pairs, HICA can be engineered to have metal affinity and thus be easily purified via IMAC

Collecting Cultural Feedback on Ethiopian Views on Girls, Sport, and Voice

This qualitative study used data collected from five interviews conducted to gather cultural and linguistic feedback on a 21-question instrument created for a future study focused on Ethiopian girl athletes. Participants met the following criteria: (a) native to Ethiopia (b) at least 15 years old and (c) proficient English reading and speaking skills. Participants did not complete the questionnaire, rather, they read it and provided cultural and linguistic feedback on its content. Participants provided feedback via interviews both in person and over the phone during a two-week period. The instrument was designed to collect data from members of Girls Gotta Run Foundation, a non-profit organization in Ethiopia that uses sport to promote gender equity. It consists of questions that focus on girls’ perceptions on the constructs of gender, sport, and voice empowerment. Findings demonstrated the importance of providing greater flexibility in questions that related to age and career goals, while the impact running has on Ethiopian culture was reinforced. Conducting this pilot study demonstrated the importance of testing cross-cultural instruments prior to final instrument administration to gain the most culturally accurate data possible

Channeling John Dewey: What Would Vermont\u27s Philosopher of Democracy Have to Say About Personalized Learning?

In John Dewey’s educational framework, the process and product are inseparable; achieving democratic ends cannot result from undemocratic means. For him, the full humanization of people depended not upon externally imposed curriculum and management systems, but rather on responding to the intrinsic needs, interests, and powers of the individual to be educated. The trend in many states on personalized learning, flexible pathways, and proficiency-based assessment, provides a foundation for transforming the conventional system of education, with its standardization, testing, and grading towards Dewey’s vision of a more socially just, inclusive, and (small d) democratic system. So, what might Dewey have to say about personalized learning as a model with the potential to revolutionize the entrenched system? This essay addresses five problem-situations and questions that might merit his consideration: The contradiction between personalization and the creation of democratic community-building; Corporate interest in personalized learning, and the ‘perils of the personalized playlist’; Shifting from individual to ecological intelligence; Challenging the ‘school-to-college-and-career’ pipeline; and Personalization and the elusive quest for equity Personalized learning is one of the most important developments in educational reform and renewal toward a more socially just, egalitarian system with the potential to engage students fully in their learning and in their communities. However, there are many pitfalls along the road to implementation, from the problem of stagnant mindsets and mental models to corporate hijacking of the discourses around personalization. This essay highlights ways that we might best avoid these snares, so that the full power of personalized learning might be realized

Administrative Effectiveness and Public Administration Practice in Midwestern State Employment Agencies

Researchers have examined the multiple dimensions of unemployment. Performance rates had declined significantly in a majority of CR 5 states in the 3-year period, 2007-2009. What was not known, however, was the effectiveness of the public administration practice in individual state employment agencies when addressing the economic crisis and rising unemployment. In this evaluative study, the effectiveness of each of 10 state employment agencies in the Central Region 5 of the U.S. Department of Labor Employment and Training Administration was investigated.https://scholarworks.waldenu.edu/archivedposters/1108/thumbnail.jp

Unleashing the Organizational Potential : Extra-Role Creativity, Innovation, and Performance in the Context of Social Influences

New ideas are important for companies to evolve and overcome problems. This dissertation focuses on utilizing the companies’ potential for these ideas: their employees. It examines in detail extra-role creativity as a subdimension of creativity. Extra-role creativity describes the discretionary generation of new and useful ideas beyond one’s job role expectations. This subdimension has, to our knowledge, only been theoretically proposed, but not empirically assessed. The dissertation further evaluates its relation to innovation—the implementation of ideas—and performance. In addition, it examines social factors relevant to fostering extra-role creativity, innovation, and performance. It specifically looks at the influences of the direct social environment of an employee exerted through leadership behavior and different team climate dimensions. Taken together, this dissertation addresses three research questions. Each manuscript relates to one or more of the following research questions: 1) How is extra-role creativity perceived by employees? 2) What is the connection between extra-role creativity and innovation as well as between extra-role creativity and performance? 3) Which leadership behaviors or team climate dimensions are relevant for extra-role creativity, innovation, and performance

Mapping Evidence From the Literature About the Experience of Internationally Educated Health Professionals to Canadian Professional Competency Profiles of Physiotherapists, Occupational Therapists, Physicians, and Pharmacists.

Abstract Purpose: To review experiences of internationally educated healthcare professionals (physiotherapists, occupational therapists, physicians and pharmacists) in their new country and to map these experiences using a competency framework. Method: Database (Medline, EMBASE, CINAHL, and PsycINFO ) were systematically searched to include articles published between 2000 and 2017 and described the experiences of internationally educated healthcare professionals (IEHPs) in physiotherapists, occupational therapists, physicians and pharmacist in attaining registration in new country. The data were synthesised using conventional content analysis. Emerging themes were mapped across a competency framework based on the profiles of the selected professions. Result: Thirteen articles were included; most were conducted in Canada and among internationally educated medical doctors. Themes were mapped to all the roles in the professional competency profiles except the Health Advocate role. Communicator role was the most frequently discussed; internationally educated healthcare professionals often needed to improve in culture-specific communication including verbal and non-verbal cues during assessment, documentation and treatment of clients. A pictorial representation was created for describing internationally educated healthcare professionals’ deficiencies in roles/ competencies for professional practice. In this representation, cultural-language and confidence deficits contribute to the deficiencies seen in roles and competencies among internationally educated healthcare professionals in their new country. Conclusion: Internationally educated healthcare professionals’ ability to fulfill the explicit roles in the competency profiles may depend largely on having good cultural-language competence as well as confidence. Exposing the internationally educated healthcare professionals to local practice is one of the strategies for cultural-language and confidence enhancement, consistent with findings reported in the nursing profession

A survey of emergency department use in patients with cyclic vomiting syndrome

<p>Abstract</p> <p>Background</p> <p>Cyclic vomiting syndrome (CVS), a chronic disorder characterized by recurrent episodes of vomiting, is frequently unrecognized and is associated with high utilization of emergency department (ED) services.</p> <p>Methods</p> <p>A web-based survey was posted on the Cyclic Vomiting Syndrome Association (CVSA) website to assess utilization of ED services in patients with CVS.</p> <p>Results</p> <p>Of 251 respondents, 104 (41.4%) were adult CVS patients and 147 (58.6%) were caregivers of pediatric and adult patients. In the adult group, the median number of ED visits for CVS symptoms was 15(range 1 - 200), with a median of 7 ED visits prior to a diagnosis of CVS (range 0 - 150). In the caregiver group, the median number of ED visits was 10 (range 1 - 175) and the median number of ED visits prior to a diagnosis of CVS was 5 (range 0 - 65). CVS was not diagnosed in the ED in 89/104 (93%) adults and 119/147 (93%) patients in the caregiver group. CVS was not recognized in the ED in 84/95 (88%) of adults and 97/122 (80%) of patients in the caregiver group, despite an established diagnosis of CVS.</p> <p>Conclusion</p> <p>There is a sub-group of adult and pediatric CVS patients who are high utilizers of ED services and CVS is not recognized in the ED in the majority of patients. Improved efforts to educate ED physicians are indicated to optimize treatment of patients with CVS and to decrease potential overuse of ED services.</p

Bim and Bmf synergize to induce apoptosis in Neisseria gonorrhoeae infection

Abstract: Bcl-2 family proteins including the pro-apoptotic BH3-only proteins are central regulators of apoptotic cell death. Here we show by a focused siRNA miniscreen that the synergistic action of the BH3-only proteins Bim and Bmf is required for apoptosis induced by infection with Neisseria gonorrhoeae (Ngo). While Bim and Bmf were associated with the cytoskeleton of healthy cells, they both were released upon Ngo infection. Loss of Bim and Bmf from the cytoskeleton fraction required the activation of Jun-N-terminal kinase-1 (JNK-1), which in turn depended on Rac-1. Depletion and inhibition of Rac-1, JNK-1, Bim, or Bmf prevented the activation of Bak and Bax and the subsequent activation of caspases. Apoptosis could be reconstituted in Bim-depleted and Bmf-depleted cells by additional silencing of antiapoptotic Mcl-1 and Bcl-XL, respectively. Our data indicate a synergistic role for both cytoskeletal-associated BH3-only proteins, Bim, and Bmf, in an apoptotic pathway leading to the clearance of Ngo-infected cells. Author Summary: A variety of physiological death signals, as well as pathological insults, trigger apoptosis, a genetically programmed form of cell death. Pathogens often induce host cell apoptosis to establish a successful infection. Neisseria gonorrhoeae (Ngo), the etiological agent of the sexually transmitted disease gonorrhoea, is a highly adapted obligate human-specific pathogen and has been shown to induce apoptosis in infected cells. Here we unveil the molecular mechanisms leading to apoptosis of infected cells. We show that Ngo-mediated apoptosis requires a special subset of proapoptotic proteins from the group of BH3-only proteins. BH3-only proteins act as stress sensors to translate toxic environmental signals to the initiation of apoptosis. In a siRNA-based miniscreen, we found Bim and Bmf, BH3-only proteins associated with the cytoskeleton, necessary to induce host cell apoptosis upon infection. Bim and Bmf inactivated different inhibitors of apoptosis and thereby induced cell death in response to infection. Our data unveil a novel pathway of infection-induced apoptosis that enhances our understanding of the mechanism by which BH3-only proteins control apoptotic cell death