An improved, optimised and robust keratin azure assay for accurate assessment of keratinase activity.

Keratin, in the form of coarse sheep wool, has been identified as an undervalued natural resource, which with the appropriate tools (e.g. a keratinase biocatalyst) can be repurposed for various textile and industrial biotechnology applications. For these purposes, we describe a novel method for identifying keratinase activity through the use of α-keratin azure (KA), an anthraquinone dyed substrate. A colourimetric method monitored the keratinase activity of Proteinase K (PK), which degrades the KA substrate and releases soluble products that are observed at 595 nm. Initially, the azure dye standard, Remazol Brilliant Blue R (RBBR), was used to calibrate the assay and allowed the kinetics of the keratinase-catalysed reaction to be determined. The assay was also used to investigate substrate pre-treatment, as well as different reaction quenching/work up conditions. Milling and washing of the KA substrate provided the best reproducibility and centrifugation was the most effective method for removing unreacted starting material. This assay was then applied to investigate the reduction of the keratin disulfide bond on keratinase-catalysed degradation. This optimised, improved and robust method will enable identification of keratinases ideally suited for application in the valorisation of the α-keratin found in natural wool fibres.

Exploring treatment decision-making in cancer management for rural residents: Patient and provider perspectives on a recently established regional radiotherapy service.

AIM: The aim of this study was to examine stakeholders' views of how the establishment of the South West Radiation Oncology Service in Bunbury, Western Australia, has affected treatment decision-making, and the extent to which patients are involved in the decision-making process. METHODS: Semi-structured in-depth interviews were undertaken with 21 service providers involved in the treatment and care of people with cancer, and 17 adults diagnosed with cancer who opted to undergo radiotherapy (RT) treatment at the Service. Data were subject to thematic analysis using the qualitative data analysis software NVivo 10. RESULTS: Patients were overwhelmingly positive about their experiences at the Service, praising the quality of the care provided and noting the convenience associated with receiving treatment locally. Most patients reported feeling involved in decision-making, but tended to rely on and generally adopt advice from medical professionals regarding type and location of treatment. Although service providers in the region had become more aware of the regional RT service and referrals had increased, some patients continued to travel to the metropolitan area for treatment. Reasons identified for this included the need for more specialized RT treatment and the more extensive range of allied health services offered. CONCLUSION: Increased convenience and a lower financial burden are key reasons why rural cancer patients prefer treatment at a regional RT service rather than travelling to metropolitan centers. These factors highlight the need for ongoing improvement in access to local RT services so that disparities in cancer outcomes between rural and metropolitan patients are reduced.