Purification of Global Regulator, Spx, and RNA Polymerase from Staphylococcus aureus for Use in In Vitro Transcription of Redox Genes

Spx is a global regulator discovered in Bacillus subtilis to suppress basal growth and development processes and activate transcription of genes involved in thiol homeostasis when a cell encounters oxidative stress. Its activity relies on reversible thiol-disulfide bond formation and binding RNA polymerase rather than DNA. The discovery that Staphylococcus aureus global virulence regulator, SarA, is more active upon cysteine reduction suggests that redox response could mediate virulence in this important human pathogen. We describe the cloning of spx from S. aureus strain RN6390, overexpression in Escherichia coli, and purification of native protein. Antibodies against Spx were raised for western analysis. Spx from S. aureus was highly active in a B. subtilis in vitro transcription system, stimulating expression of trxB, the gene encoding thioredoxin reductase, without reducing agents. RNA polymerase was partially purified from S. aureus, and the enzyme was active, catalyzing transcription of rpsD, but not trxB

Purification of Global Regulator, Spx, and RNA Polymerase from Staphylococcus aureus for Use in In Vitro Transcription of Redox Genes

Spx is a global regulator discovered in Bacillus subtilis to suppress basal growth and development processes and activate transcription of genes involved in thiol homeostasis when a cell encounters oxidative stress. Its activity relies on reversible thiol-disulfide bond formation and binding RNA polymerase rather than DNA. The discovery that Staphylococcus aureus global virulence regulator, SarA, is more active upon cysteine reduction suggests that redox response could mediate virulence in this important human pathogen. We describe the cloning of spx from S. aureus strain RN6390, overexpression in Escherichia coli, and purification of native protein. Antibodies against Spx were raised for western analysis. Spx from S. aureus was highly active in a B. subtilis in vitro transcription system, stimulating expression of trxB, the gene encoding thioredoxin reductase, without reducing agents. RNA polymerase was partially purified from S. aureus, and the enzyme was active, catalyzing transcription of rpsD, but not trxB

Does an Online Clinical Educator Preparation and Support Program Change Practice?

Preparation of clinicians to act as student placement supervisors is important to ensure quality student placements for the development of the skills needed for competent performance in the workplace. Clinical educator preparation programs are offered in many formats, but these programs are rarely evaluated for impact on practice. In this article, we describe the results of the evaluation of an online clinical educator preparation and support (CEPS) program. Thirty allied health professionals, across a range of professions, responded to a survey regarding their experience of the program, usage patterns and their application of learning to practice. As a result of participation in the program, there was a significant increase in confidence levels in a number of topic areas covered in the program, and a quarter of respondents had changed their student supervision practices as a result of participation. Due to a low response rate at the three month follow-up survey, planned interviews to explore the impact of change in practice on the student placement experience could not be completed. While the study was not able to measure the impact of the CEPS program on placement quality, it did show that the CEPS program is able to significantly increase supervisor confidence in a number of areas, and is able to effect change in practice

Parenting during a Pandemic: Mothers and Disabled Children in Aotearoa/New Zealand—A Hidden Minority

Every country has its own COVID-19 pandemic story; similarly, every family has their own experiences of lockdowns, isolation, illness, death, struggles, and resiliency related to the pandemic. Although myriad narratives appear about these familial and societal experiences, few explore those of mothers of disabled children; these have been largely invisible, and as a result, this minority group and their needs have failed to be addressed by those who make decisions and plan for public health crises and for the subsequent recovery. Autoethnography, a qualitative method that coalesces personal experience and research literature to advance sociological understanding, underpins this exploration. The authors are New Zealand/Aotearoa mothers of disabled children. Our approach employs autoethnographic reflection about our pandemic experiences to create mean-ing, forge identities, and explore power structures. Connections of our family stories enable the creation of an understanding of what has happened in our communities. The authors’ reflections on their pandemic experiences are woven together with stories of how governments, schools, public health organizations, disability organizations, healthcare providers and communities directed us and responded to or failed to address our needs. We have identified five interwoven themes throughout our stories: anxiety, invisibility, devalued lives, coping, and advocacy. Together, as an outcome of the autoethnographic study of our pandemic experiences, we offer ideas for survival to pass on to mothers for future disasters and crises. Furthermore, we have developed recommendations for organizations and others living with disability

An Investigation of Increased Rates of Autism in U.S. Public Schools

This study is intended to enhance our understanding of the dramatic increase in autism prevalence rates across the United States. A robust national and multistate sample of children and youth (5–22 years of age) was gathered and analyzed, using visual and statistical analysis of autism eligibility over a 15‐ to 17‐year period. Although the impact of environmental or genetic influences cannot be entirely ruled out, we identified significant shifts in eligibility trends that substantially contribute to the remarkable increase in autism prevalence. Assessment procedures and criteria for autism have sustained an indelible influence from this diagnostic migration, which has had a lasting impact on public schools. We use this information to provide meaningful implications for practicing psychology in the schools

The Role of Discrete Emotions in Job Satisfaction: A Meta-Analysis

[Summary] The relationship between emotions and job satisfaction is widely acknowledged via affective events theory (AET). Despite its widespread use, AET was not designed to address why specific emotions might differentially relate to job satisfaction. We utilize appraisal theory of emotion to refine AET and provide this nuanced theorizing. We meta‐analytically test our ideas with 235 samples across 99 883 individuals and 22 600 intra‐individual episodes. We test two approaches—specific emotion experiences (16 discrete emotions) versus general emotion experiences (positive or negative emotions)—and present empirical evidence of their similarities and differences with job satisfaction. Our findings suggest that specific emotions with circumstance‐agency appraisals (e.g., depression and happiness) have the strongest associations with job satisfaction compared to emotions with self‐ and other‐agency appraisals and general emotion experiences. However, more variability is observed for negative emotions and job satisfaction compared to positive emotions. Further, we address and even challenge influential critiques of emotions and job satisfaction via a meta‐analytic test of five moderators—emotion intensity versus frequency, target of emotion, job satisfaction measure, level of analysis, and time referent for emotion and job satisfaction recall. In sum, we advance academic and practitioner understanding of the relationship between emotions and job satisfaction

Co‑designed, culturally tailored cervical screening education with migrant and refugee women in Australia : a feasibility study

Background: Participation of culturally and linguistically diverse (CALD) women from migrant and refugee backgrounds in cervical screening is crucial to eliminate cervical cancer as a public health problem within the next 20 years. However, CALD women report low participation in cervical screening. Barriers to participation can be addressed with culturally tailored, community-based programs. There is a need for research to explore the process, feasibility, acceptability and barriers to cultural tailoring in the delivery and evaluation of cervical screening health education. Methods: CALD community health workers took part in a 2 day training program then co-designed, culturally tailored and co-facilitated cervical screening health promotion forums within their communities. Forums were delivered to a total of seven groups, involving 12 sessions and 71 CALD women. The forums were evaluated for feasibility, acceptability, implementation and effectiveness using a survey, interviews and observations. Data were collected from CALD women, facilitators and researchers. Results: The co-design and co-delivery of cervical screening health promotion forums was time and resource intensive however allowed for deeper cultural tailoring resulting in engagement with ‘hard to reach’ CALD women, improved health literacy and intention to screen. Flexibility in the intervention implementation was crucial to ensure forums were responsive to community interests and needs. Online delivery of the forums in response to the COVID-19 pandemic was acceptable to most groups. Conclusions: Co-designed, culturally tailored cervical screening health promotion forums are feasible and acceptable to CALD women, in both face-to-face and online formats. Adjustments to the intervention protocol were recommended to improve future implementation

PenQuest Volume 5, Number 1

Table of Contents for this Volume: Success by Shatney Maria by Jane O’Neal Intrusions by Mark McBride The Mystery of the Back Porch Light by Nature Johnston Truth and the Violin by Shatney Corporate America by Julie Crowell Pete’s Cafe by Nature Johnston Geranium by Anne Benjamin The Man Who Buried His Books by William Slaughter Erasures by William Slaughter Mind You and other poems by Kate Mathews Coffee in the Tea Room by Kathleen O’Brien The Children by Katharine Rodier Sisters, Reclamation, Not Wanting to Say, “I Told You So,” But… by Kathleen O’Brien Genetics by Kathleen O’Brien The Anguish of Flames by Kathleen O’Brien turning plows by Mark McBride A Valediction for My Father by Jonathan Williams Untitled by Mark Sablow Artificial Portrait by Kevin Christenson Untitled by Latrell Mickler Untitled by Kevin Christenson Galvanistic Ascension by Mark Grisham Power Surge by Mark Grisham Untitled by Lori Kirsbau