A polar filter in DNA polymerases prevents ribonucleotide incorporation.

The presence of ribonucleotides in DNA can lead to genomic instability and cellular lethality. To prevent adventitious rNTP incorporation, the majority of the DNA polymerases (dPols) possess a steric filter. The dPol named MsDpo4 (Mycobacterium smegmatis) naturally lacks this steric filter and hence is capable of rNTP addition. The introduction of the steric filter in MsDpo4 did not result in complete abrogation of the ability of this enzyme to incorporate ribonucleotides. In comparison, DNA polymerase IV (PolIV) from Escherichia coli exhibited stringent selection for deoxyribonucleotides. A comparison of MsDpo4 and PolIV led to the discovery of an additional polar filter responsible for sugar selectivity. Thr43 represents the filter in PolIV and this residue forms interactions with the incoming nucleotide to draw it closer to the enzyme surface. As a result, the 2'-OH in rNTPs will clash with the enzyme surface, and therefore ribonucleotides cannot be accommodated in the active site in a conformation compatible with productive catalysis. The substitution of the equivalent residue in MsDpo4-Cys47, with Thr led to a drastic reduction in the ability of the mycobacterial enzyme to incorporate rNTPs. Overall, our studies evince that the polar filter serves to prevent ribonucleotide incorporation by dPols.

Reflective practice: a framework for case manager development.

PURPOSE/OBJECTIVES: The role of a nurse case manager (NCM) incorporates practice that is built upon knowledge gained in other roles as well as components unique to case management. The concept of reflective practice was used in creating a framework to recognize the developmental stages that occur within community based case management practice. The formation of this framework and its uses are described in this article. PRIMARY PRACTICE SETTING: The practice setting is a community based case management department in a large midwestern metropolitan health care system with Magnet recognition. Advanced practice nurses provide care for clients with chronic health conditions. FINDINGS/CONCLUSIONS: Twenty-four narratives were used to identify behaviors of community based case managers and to distinguish stages of practice. The behaviors of advanced practice found within the narratives were labeled and analyzed for similarities. Related behaviors were grouped and descriptor statements were written. These statements grouped into 3 domains of practice: relationship/partnership, coordination/collaboration, and clinical knowledge/decision making. The statements in each domain showed practice variations from competent to expert, and 3 stages were determined. Reliability and validity of the framework involved analysis of additional narratives. The reflective practice process, used for monthly case review presentations, provides opportunity for professional development and group learning focused on improving case manager practice. The framework is also being used in orientation as new case managers acclimate to the role. IMPLICATIONS FOR CASE MANAGEMENT PRACTICE: Reflective writing has unveiled the richness and depth of nurse case manager practice. The depth of knowledge and skills involved in community-based case management is captured within this reflective practice framework. This framework provides a format for describing community based case manager practice development over the course of time and has been used as a tool for orientation and peer review.