Establishing a Biochemical System for the Purification and ATPase activity of GST-Dbp5

The export of mRNA out of the nucleus is a crucial step for eukaryotic gene expression. The export of mRNA transcripts is aided by Mex67, which allows export through the nuclear pore complex doorways in the nuclear envelope. Once out of the nucleus, a protein known as Dbp5, bound to ATP, Gle1, and Nup42 aids in the directionality of mRNA export by helping remove Mex67 from the mRNA strand. Following interaction with RNA, Dbp5 then hydrolyzes ATP so that it unbinds the mRNA, allowing for enzyme recycling. Previous efforts worked towards the purification of Dbp5, but the attempts were unsuccessful due to low expression of recombinant protein in E.coli. In this project, I am focusing on enhancing the bacterial induction in order to establish robust purification of recombinant Dbp5. This will help in developing ATPase assays involving Dbp5, Nup42, and Gle1. These ATPase assays will aid in better understanding the effects of Nup42 and Gle1 on Dbp5’s ATPase activity and will allow for future study on Dbp5’s ATPase activity. In order to enhance the bacterial induction, E. coli cells were transformed with a GST-Dbp5 plasmid and were induced with varying amounts of IPTG to determine the best procedure for bacterial induction. Results from the bacterial induction have indicated that alternative methods for bacterial induction should be explored. Future experiments will look into further enhancing the bacterial induction of Dbp5 in order to establish a biochemical system analyzing the ATPase activity of GST-Dbp5

Practice and research in Australian massage therapy: a national workforce survey

Background: Massage is the largest complementary medicine profession in Australia, in terms of public utilisation, practitioner distribution, and number of practitioners, and is being increasingly integrated into the Australian health care system. However, despite the increasing importance of massage therapists in Australian health care delivery, or the increased practice and education obligations this may entail, there has been little exploration of practice, research, and education characteristics of the Australian massage therapist workforce.Purpose: To identify practice, research, and education characteristics among the Australian massage therapist workforce.Settings: The Australian massage therapy profession.Participants: 301 randomly selected members of the Association of Massage Therapists (Australia).Research Design: A 15-item, cross-sectional telephone survey.Main Outcomes Measures: Massage therapists’ demographic information, practice characteristics, and education and research characteristics.Results: Most respondents (73.8%) worked 20 hours per week or less practising massage, nearly half of all respondents (46.8%) treated fewer than 10 massage clients per week, and over three-quarters (81.7%) of respondents were self-employed. Massage therapy was the sole source of income for just over half (55.0%) of the study respondents. Only 5.7% of respondents earned over the average wage ($50,000) through their massage activities. Nearly half of all respondents (43.3%) reported regularly exceeding their continuing professional education (CPE) quota mandated by their professional association. However, 21.1% reported struggling to achieve their CPE quota each year. Over one-third of respondents (35.6%) were not interested in acquiring further CPE points beyond minimum requirements. Respondents were significantly more likely to have an active approach to research if they had higher income (p = .015). Multivariate analysis showed factors associated with access to CPE to be the only significant predictors for increased CPE.Conclusions: The massage profession in Australia remains largely part-time and practitioners earn less than the average Australian wage. The factors that underlie research and education involvement appear to be highly individualised and, therefore, policies targeting specific groups may be arbitrary and ineffective

Establishing a Biochemical System for the Purification of GST-Dbp5 to assess ATPase activity

The export of mRNA out of the nucleus, the site of its production, and to the cytoplasm, where proteins are translated, is a crucial step for eukaryotic gene expression. This is aided by Mex67 binding to the transcript, which allows export through the nuclear pore complexes (NPCs), selective doorways embedded in the nuclear envelope. Once the transcript reaches the cytoplasm, a protein known as Dbp5, bound to ATP, Gle1, and Nup42 aids in the directionality of mRNA export by binding to the mRNA strand and removing Mex67. Following interaction with RNA, Dbp5 then hydrolyzes ATP so that it unbinds the mRNA, allowing for enzyme recycling. Therefore, Dbp5 ATPase activity is essential for mRNA export and cell viability. Factors that influence this activity can be accessed through biochemical purification and in vitro analysis. The goal of this work has been to enhance purification of recombinant Dbp5 for this analysis. Specifically, I have focused on optimizing induction of GST-Dbp5 in E. coli Rosetta cells. These efforts have yielded successful induction of Dbp5 at 16°C, and ongoing experiments are aimed at biochemical purification from induced cells. Future experiments will work towards establishing a biochemical system to analyze the ATPase activity of GST-Dbp5

Diagnosis and Management of Depression in Three Countries: Results from a Clinical Vignette Factorial Experiment

Abstract Objective: International differences in disease prevalence rates are often reported and thought to reflect different lifestyles, genetics, or cultural differences in care-seeking behavior. However, they may also be produced by differences among health care systems. We sought to investigate variation in the diagnosis and management of a "patient," with exactly the same symptoms indicative of depression, in three different health care systems (Germany, the United Kingdom, and the United States). Method: A factorial experiment was conducted in which 384 randomly selected primary care physicians viewed a video vignette of a "patient" presenting with symptoms suggestive of depression. Results: Most physicians listed depression as one of their diagnoses, but German physicians were more likely to diagnose depression in women, while British and American physicians were more likely to diagnose depression in men (p=.0251). American physicians were almost twice as likely to prescribe an anti-depressant (p=.0241) as British physicians. German physicians were significantly more likely to refer the patient to a mental health professional (p<.0001) than British or American physicians. German physicians wanted to see the patient in follow-up sooner (p<.0001) than British or American physicians. Conclusions: Primary care physicians in different countries diagnose exactly same symptoms of depression differently, depending on the patient's gender. There are also significant differences between countries in the management of a "patient" with symptoms suggestive of depression. International differences in prevalence rates for 3 depression, and perhaps other diseases, may in part result from differences among health care systems in different countries

Nonelective coronary artery bypass graft outcomes are adversely impacted by Coronavirus disease 2019 infection, but not altered processes of care: A National COVID Cohort Collaborative and National Surgery Quality Improvement Program analysisCentral MessagePerspective

Objective: The effects of Coronavirus disease 2019 (COVID-19) infection and altered processes of care on nonelective coronary artery bypass grafting (CABG) outcomes remain unknown. We hypothesized that patients with COVID-19 infection would have longer hospital lengths of stay and greater mortality compared with COVID-negative patients, but that these outcomes would not differ between COVID-negative and pre-COVID controls. Methods: The National COVID Cohort Collaborative 2020-2022 was queried for adult patients undergoing CABG. Patients were divided into COVID-negative, COVID-active, and COVID-convalescent groups. Pre-COVID control patients were drawn from the National Surgical Quality Improvement Program database. Adjusted analysis of the 3 COVID groups was performed via generalized linear models. Results: A total of 17,293 patients underwent nonelective CABG, including 16,252 COVID-negative, 127 COVID-active, 367 COVID-convalescent, and 2254 pre-COVID patients. Compared to pre-COVID patients, COVID-negative patients had no difference in mortality, whereas COVID-active patients experienced increased mortality. Mortality and pneumonia were higher in COVID-active patients compared to COVID-negative and COVID-convalescent patients. Adjusted analysis demonstrated that COVID-active patients had higher in-hospital mortality, 30- and 90-day mortality, and pneumonia compared to COVID-negative patients. COVID-convalescent patients had a shorter length of stay but a higher rate of renal impairment. Conclusions: Traditional care processes were altered during the COVID-19 pandemic. Our data show that nonelective CABG in patients with active COVID-19 is associated with significantly increased rates of mortality and pneumonia. The equivalent mortality in COVID-negative and pre-COVID patients suggests that pandemic-associated changes in processes of care did not impact CABG outcomes. Additional research into optimal timing of CABG after COVID infection is warranted