Approaches for Conjugating Tailor-Made Polymers to Proteins.

A series of methods are outlined for attaching functional polymers to proteins. Polymers with good control over structure, functionality, and composition can be created using reversible addition-fragmentation chain transfer (RAFT) polymerization. These polymers can be covalently linked to enzymes and proteins using either the "grafting-to" approach, where a preformed polymer is attached to the protein surface, or the "grafting-from" approach, where the polymer is grown from the protein surface. Methods for grafting-to, or attaching the RAFT chain transfer agent to the protein surface outlined include the commonly used carbodiimide/activated ester (EDC/NHS) coupling. Methods are also outlined to graft-from the surface of the protein using RAFT polymerization. Additionally, it is possible to site specifically introduce a reactive azide group to the protein surface using enzymatic ligation as a posttranslational modification. This reactive azide group can be conjugated to an alkyne-containing polymer using highly efficient click chemistry. These robust protocols can produce protein-polymer conjugates with various architectures and functionalities. Methods are also outlined for characterization of the resulting bioconjugates.

Strategies for Biophysical Characterization of Protein-Polymer Conjugates.

Protein-polymer conjugates are increasingly viewed as promising avenues to producing industrial enzymes with high activity capable of withstanding potentially harsh reaction conditions, or to designing novel therapeutics with triggered release, controlled masking, or increased resistance to proteolytic degradation. Common among these applications are the desire to improve the stability of protein-polymer conjugates to unfolding by exposure to chemicals or thermal stress. Thus, assays that allow researchers to robustly and easily characterize protein-polymer conjugates by obtaining thermodynamic parameters for folding stand to play an important role in the development of improved protein-polymer conjugates. Herein, we describe two techniques, differential scanning fluorimetry and intrinsic tryptophan fluorescence, used in our laboratories to obtain thermodynamic parameters of unfolding that allow for direct comparison of protein-polymer conjugates and the myriad effects of variations in attachment site, polymer identity, and polymer length. These two experiments, which are easily amenable to parallelization, are presented as high-throughput replacements for more traditionally employed circular dichroism experiments and as complements to functional chemical stability or functional thermal stability experiments. Each assay is presented in a parallelized format that allows for rapid scaling and high-throughput analysis of protein-polymer conjugate libraries. Descriptions of the assays include a discussion of advantages and disadvantages alongside protocol details and approaches to data analysis.

Patient journeys: the process of clinical redesign.

*Clinical process redesign is a successful improvement method that has been used to increase access to health services in 60 public hospitals across New South Wales, and at Flinders Medical Centre (FMC) in South Australia. *The method focuses on the patient journey as the primary improvement locus, and uses process mapping to identify the value-adding steps in that journey; it involves redesign teams identifying and eliminating non-value-adding steps to improve flow and reduce delays in access to emergency and elective care. *The method engages clinicians, managers, patients and carers, and delivers real gains in health care delivery. *This article outlines the clinical process redesign programs being used by NSW Health and at FMC.

Redesigning care at the Flinders Medical Centre: clinical process redesign using "lean thinking".

*The Flinders Medical Centre (FMC) Redesigning Care program began in November 2003; it is a hospital-wide process improvement program applying an approach called "lean thinking" (developed in the manufacturing sector) to health care. *To date, the FMC has involved hundreds of staff from all areas of the hospital in a wide variety of process redesign activities. *The initial focus of the program was on improving the flow of patients through the emergency department, but the program quickly spread to involve the redesign of managing medical and surgical patients throughout the hospital, and to improving major support services. *The program has fallen into three main phases, each of which is described in this article: "getting the knowledge"; "stabilising high-volume flows"; and "standardising and sustaining". *Results to date show that the Redesigning Care program has enabled the hospital to provide safer and more accessible care during a period of growth in demand.

Coping with prostate cancer: a quantitative analysis using a new instrument, the centre for clinical excellence in urological research coping with cancer instrument.

OBJECTIVES: To assess the reliability (internal and interrater) and validity (concurrent) of a new interview measure for assessing patients' ability to cope with cancer, the Centre for Clinical Excellence in Urological Research Coping with Cancer Instrument (CCCI), and to determine whether there is an underlying structure to the various coping strategies used by patients with prostate cancer. METHODS: Eighty patients with prostate cancer were interviewed using the CCCI. The participants also completed measures of quality of life and anxiety and depression. RESULTS: The psychometric properties of the CCCI were acceptable. Factor analysis revealed that coping with prostate cancer can be described along five dimensions: positive problem solving (fighting against the illness, seeking information); self-reliance (developing a lay explanation, distrusting doctors); emotional availability (not withdrawing from others); distress (brooding, self-pity); and solace (taking alcohol or drugs to improve mood). These coping styles were correlated with age, quality of life, self-reported prostate-specific antigen level, and measures of anxiety and depression. CONCLUSIONS: The results of the present study have led to a greater understanding of the underlying coping styles used by patients with prostate cancer. Although some of these are similar to those found in other cancer populations, others, such as self-reliance and solace, represent unique and potentially clinically significant responses to prostate cancer diagnosis and treatment. A larger scale longitudinal study is needed to determine the wider clinical implications associated with each coping style.