The effects of DNA sequence on interferon mRNA biosynthesis and stability in E. coli

The aim of this investigation was determine the effect of DNA sequences on interferon mRNA biosynthesis and stability in E.coli. Interferon al and a2 DNA sequences are 80% homologous. Despite this large degree of similarity different levels of protein and different transcription products were obtained when the two genes were cloned into identical vectors. Increasing the plasmid copy number and introducing a transcription terminator immediately downstream from the interferon sequence increased al production. Deleting the sequence upstream from the -35 region of the promoter or reducing the length of the 3' non-coding region had little effect on a2 production. Highest levels of a2 interferon were obtained by altering the ribosome binding site. This necessitated decreasing the plasmid copy number to achieve maximum plasmid stability. In contrast, all al containing plasmids were stability maintained in the bacteria. Analysis of the transcription products of all the interferon producing clones revealed that multiple mRNA species were generated by all the constructs except those which contained the termination signal. Removal of the sequence 5' to the -35 region of the promotor resulted in reduced transcription, while shortening the 3' non-coding region lowered termination efficiency within the remaining sequence facilitating the production of polycistronic messages. Stability studies were undertaken to determine the half-lives of the mRNA species. Most decayed with a half-life of 1-1.5 minutes. However, more stable species were produced by the constructs containing the termination signal. Comparison of the mRNA species produced by Similar al and a2 producing strains revealed that while all the al and a2 mRNA were of sufficient length for translation to yield mature interferon three a2 mRNA species appeared too short. These prematurely terminating mRNA were further characterized. The smallest terminated at a rho-dependant termination site while the other two species terminated at rho-independant sites. Termination at the rho-dependant site could be eliminated if the rare arginine codons AGG present in the early coding sequence were replaced by the more commonly used arginine codons CGT. Transcription profiles and mRNA half-lives were determined in different media at different growth rates to see whether production of the prematurely terminating species was affected. While all species were produced under all conditions the distribution of mRNA species altered with both media and growth phase.<p

Attitudes towards sub-domains of professionalism in medical education: defining social accountability in the globalizing world

Background: Unmet health needs of populations around the world are a major contributor to lagging health outcomes globally. Medical professionals have a duty to address the health needs of their communities. In a globalizing world, the needs may seem limitless. Yet, most training involves immersion in one health system and its resources. How do practitioners reconcile this potentially limitless demand with their focused training and in understanding their duty to the populations they serve?Methods: A mixed-method design was used. We distributing a pre-validated survey examining all facets of professionalism to the Department of Family Medicine at the University of Ottawa. This was followed by interviewing a purposive sample of residents and faculty with different levels of interest in working with underserved populations, to examine attitudes towards social accountability.Results: Quantitative results did not replicate the factor structure of the pre-validated survey in our cohort. This and other gaps in individual responses were used to construct an interview guide. Interviews revealed differences between residents and faculty. Residents were likely to see social accountability as flowing from personal interest as opposed to a professional duty; and residents’ sense of duty can be met through good care of individual patients under their sphere of care. Faculty were more likely to discuss facets of care that they could influence at the health system level nationally and beyond. Conclusion: More usable and succinct instruments are needed to capture individual attitudes on social accountability. Our results identify how new physicians in family medicine in Ottawa, Canada wish to apply learning in global health to local needs, responding to the call to “think global, act local.

Deaths from stroke in US young adults, 1989-2009.

ObjectiveTo determine what the trends in stroke mortality have been over 2 decades in young adults.MethodsIn this cohort study, we analyzed death certificate data for ischemic and hemorrhagic stroke (intracerebral hemorrhage [ICH] and subarachnoid hemorrhage [SAH]) in adults aged 20-44 in the United States for 1989 through 2009, covering approximately 2.2 billion person-years. Poisson regression was used to calculate and compare time trend data between groups and to compare trends in young adults to those in adults over age 45.ResultsMortality from stroke in young adults declined by 35% over the study period, with reductions in all 3 stroke subtypes (ischemic stroke decreased by 15%, ICH by 47%, and SAH by 50%). Black race was a risk factor for all 3 stroke subtypes (relative risk 2.4 for ischemic stroke, 4.0 for ICH, and 2.1 for SAH), but declines in all stroke subtypes were more dramatic in black compared to white participants (p &lt; 0.001 for all stroke subtypes).ConclusionsAlthough hospitalizations for stroke in young patients have been increasing, the apparent decrease in mortality rates and in racial disparities suggests that recognition and treatment in this group may be improving

Simulation education: challenges and suggested practices as nursing faculty members’ experience in the Malaysian nursing curriculum

INTRODUCTION: The benefits of utilizing simulation in nursing education to mimic clinical environments are well known. Simulation education has been readily embraced and is increasingly expanding as an innovative approach to the teaching and learning process involved in preparing nursing students for clinical placement. To date, little of the research specifically addresses, in the Malaysian context, the experiences of nursing faculty members (NFMs) in terms of their preparation for the use of this approach. Objective: The aim was to explore NFMs’ experiences of simulation education implemented in the curriculum. Methodology: This study adopted a qualitative design. Focus group discussions were conducted with 23 NFMs in Malaysia and recruited using purposeful sampling. The participants took part in focus group discussions and audio-recorded interviews. The data were analysed using the NVIVO software and the thematic analysis approach. The data analysis indicated they valued its positive impact on learning outcomes, the student experience and the ability to simulate more complex scenarios during training. Results: Inadequate training, lack of resources and lack of prior planning, negatively influenced the intention to use and effectively implement simulation in the curriculum. Discussion & Conclusion The study indicated that NFMs perceived simulation education as a learning pedagogy that can improve students' learning if used effectively. They believed that to realise the potential of simulation, more support should be provided through training, the availability of necessary resources, and improved planning and organisatio

Tetrahydrophthalic Anhydrides as Addition Curing Polyimide End Caps: Thermal Isomerization of Methylendianiline 3,6-Diphenyltetrahydrophthalic Bisimides

In depth NMR studies confirm that heating a 1:2 mixture of cis, cis, cis 3,6-diphenyltetrahydrophthalic anhydride (end cap 9c) with methylenedianiline at 316 C initially yields the corresponding highly congested cis, cis, cis 3,6-diphenyltetrahydrophthalic bisimide 11, which is converted at this temperature to the observed product, the less hindered trans, cis, trans isomer 12