Use of cytology fluid samples for predictive biomarker testing in lung cancer patients.

OBJECTIVE: To provide a method of directly using cytology fluid samples for predictive biomarker testing in lung cancer patients and to determine the efficacy of a variety of fluid sample types. METHOD: A review of our in-house data from a range of cytology samples including endobronchial ultrasound (EBUS) fine-needle aspirate (FNA) needle washings (NW) and serous effusions tested on the Biocartis Idylla platform. All fluid samples were originally tested using Sanger sequencing. RESULTS: Using our method for fluid samples all of our cytology samples tested for epithelial growth factor receptor (EGFR) yielded valid results on this platform and all variant cases identified. The data showed serous fluids provided the best quality DNA, and variant genotype reports were obtained within 150 minutes. CONCLUSION: Cytology fluid samples can be used for predictive biomarker testing for lung cancer patients to provide in-house results with all fluids providing good-quality DNA.

Use of the Idylla EGFR Mutation Test for Variant Detection in Non-Small Cell Lung Cancer Samples.

OBJECTIVES: Stratified management of patients with non-small cell lung carcinoma (NSCLC) through epithelial growth factor receptor (EGFR) variant analysis has become standard clinical practice. The Idylla system is a fully compliant European in vitro diagnostics device, a fully automated platform designed to rapidly genotype formalin-fixed paraffin-embedded tissue samples. This retrospective study aims to validate the Idylla EGFR Mutation Test for use with extracted DNA from known NSCLC samples. METHODS: In this study, 20 µL of archival extracted DNA was placed directly inside the Idylla EGFR assay single-use cartridge. Idylla results were compared with the original Sanger sequencing reference method. RESULTS: The Idylla EGFR Mutation Test yielded valid results for all samples tested, confirming the variants identified by the reference method that lay within the Idylla target range. No false-positive cases were noted with the Idylla assay. Variant genotype reports were obtained within 150 minutes. CONCLUSIONS: The Idylla EGFR assay is sensitive for extracted DNA and can be reliably applied to cytologic specimens, enabling its implementation as an ancillary first-line test for patients with NSCLC.

Mediator Subunits MED16, MED14, and MED2 Are Required for Activation of ABRE-Dependent Transcription in Arabidopsis.

The Mediator complex controls transcription of most eukaryotic genes with individual subunits required for the control of particular gene regulons in response to various perturbations. In this study, we reveal the roles of the plant Mediator subunits MED16, MED14, and MED2 in regulating transcription in response to the phytohormone abscisic acid (ABA) and we determine which cis elements are under their control. Using synthetic promoter reporters we established an effective system for testing relationships between subunits and specific cis-acting motifs in protoplasts. Our results demonstrate that MED16, MED14, and MED2 are required for the full transcriptional activation by ABA of promoters containing both the ABRE (ABA-responsive element) and DRE (drought-responsive element). Using synthetic promoter motif concatamers, we showed that ABA-responsive activation of the ABRE but not the DRE motif was dependent on these three Mediator subunits. Furthermore, the three subunits were required for the control of water loss from leaves but played no role in ABA-dependent growth inhibition, highlighting specificity in their functions. Our results identify new roles for three Mediator subunits, provide a direct demonstration of their function and highlight that our experimental approach can be utilized to identify the function of subunits of plant transcriptional regulators.

Evolution of a rapid onsite evaluation (ROSE) service for endobronchial ultrasound guided (EBUS) fine needle aspiration (FNA) cytology in a UK Hospital: A 7 year audit.

OBJECTIVE: Endobronchial ultrasound fine needle aspiration (EBUS FNA) is a well-established procedure for the diagnosis and staging of lung cancer. We review our provision of this service at the Royal Devon and Exeter NHS Foundation Trust and the role of rapid onsite evaluation (ROSE) with the increasing demand for molecular markers in this era of personalized medicine. METHODS: A review of the changes in the Endoscopy clinic over the 7 years from the introduction of EBUS at the end of 2010 until 2017 was carried out. This included the availability of material obtained for diagnosis, accurate subtyping, and molecular testing. We also assessed the success of molecular genetics DNA techniques from EBUS material versus formalin fixed paraffin embedded tissue (FFPE). RESULTS: A total of 1218 EBUS cases with ROSE were reported between 2011 and 2017 Percentage diagnostic rates were calculated as 83, 82, 84, 92, 93, 94, and 92 for 2011, 2012, 2013, 2014, 2015, 2016, and 2017, respectively. Availability of material for immunocytochemistry ranged from 86 to 100% over the 7 years. Molecular testing was successfully performed for EGFR in 89-100% of requested cases and ALK testing in 87-100% of requested cases. EBUS sourced material gave on average twice the amount of DNA and fewer amplicon repeats per patient compared to FFPE material. CONCLUSION: ROSE at EBUS FNA provides access to suitable material for molecular testing with increased yields in the form of needle washings for EGFR with FFPE materials for ALK and PDL1 testing.

Health care providers' and patients' perspectives on care in HIV ambulatory clinics across Ontario.

This descriptive study represents one component of a larger project that examined the perceptions of current and best-care practices in HIV ambulatory clinics across Ontario by health care providers and patients living with HIV/AIDS. Focus groups were held with providers and patients at eight clinics. Results showed that providers' and patients' perceptions were similar. Participants were able to describe current care practices and identify two elements of best care: patient-focused care and access to care. However, both health care providers and patients acknowledged that financial constraints, appointment scheduling, and distance to clinics were some of the barriers to achieving best care. Case management and shared-care schemes are two strategies that are proposed to meet the challenge of providing collaborative integrated care that is accessible and equal to all, while still maintaining positive patient outcomes.

The nature of multidisciplinary HIV care in the ambulatory setting: a synthesis of the literature.

The purpose of this article is to provide a critical appraisal and synthesis of the literature relating to the provision of multidisciplinary HIV care in the ambulatory setting. As more people are living with HIV infection, and living longer, the demand for chronic care is great. Knowing what optimal care and treatment for people living with HIV/AIDS is and how to provide that care is an ongoing challenge. The literature suggests that the provision of care for HIV-infected individuals varies across and within HIV/AIDS clinics. This variability in care practices may influence the health and well-being of patients, and negatively impact on access to such care by people with HIV/AIDS. The goal of reviewing the literature on HIV-ambulatory clinics was to provide a description of a typical clinic, its staffing components, the care practices carried out at such a clinic (and by whom), and some barriers to providing optimal care. The ultimate goal of this review is to establish what is currently known about the provision of these services so that this information may build on existing knowledge and thereby standards of care for HIV-ambulatory clinics can be developed that could be implemented on a wide-scale basis.

Achieving best practice in routine observation of hospital patients.

This article reviews the evidence supporting best practice when undertaking patient observations in a district general hospital. Correct monitoring of the patients' progress is essential in the detection of adverse events and can potentially prevent admission to intensive care or high dependency units.

Improving policy and practice in the prevention of pressure ulcers.

This article reports on a project to update policy and practice in pressure-ulcer prevention in a district general hospital. A literature review was undertaken on pressure-ulcer prevention to identify the best methods for use in a district general hospital and to bring practice in line with current evidence. The literature review covered evidence-based tools for risk assessment, treatment protocols, policy, and patient information. An analysis of the training and educational needs of staff within the trust was also undertaken.