One-Step Preservation of Phosphoproteins and Tissue Morphology at Room Temperature for Diagnostic and Research Specimens

BACKGROUND: There is an urgent need to measure phosphorylated cell signaling proteins in cancer tissue for the individualization of molecular targeted kinase inhibitor therapy. However, phosphoproteins fluctuate rapidly following tissue procurement. Snap-freezing preserves phosphoproteins, but is unavailable in most clinics and compromises diagnostic morphology. Formalin fixation preserves tissue histomorphology, but penetrates tissue slowly, and is unsuitable for stabilizing phosphoproteins. We originated and evaluated a novel one-step biomarker and histology preservative (BHP) chemistry that stabilizes signaling protein phosphorylation and retains formalin-like tissue histomorphology with equivalent immunohistochemistry in a single paraffin block. RESULTS: Total protein yield extracted from BHP-fixed, routine paraffin-embedded mouse liver was 100% compared to snap-frozen tissue. The abundance of 14 phosphorylated proteins was found to be stable over extended fixation times in BHP fixed paraffin embedded human colon mucosa. Compared to matched snap-frozen tissue, 8 phosphoproteins were equally preserved in mouse liver, while AMPKβ1 Ser108 was slightly elevated after BHP fixation. More than 25 tissues from mouse, cat and human specimens were evaluated for preservation of histomorphology. Selected tissues were evaluated in a multi-site, independent pathology review. Tissue fixed with BHP showed equivalent preservation of cytoplasmic and membrane cytomorphology, with significantly better nuclear chromatin preservation by BHP compared to formalin. Immunohistochemical staining of 13 non-phosphorylated proteins, including estrogen receptor alpha, progesterone receptor, Ki-67 and Her2, was equal to or stronger in BHP compared to formalin. BHP demonstrated significantly improved immunohistochemical detection of phosphorylated proteins ERK Thr202/Tyr204, GSK3-α/β Ser21/Ser9, p38-MAPK Thr180/Tyr182, eIF4G Ser1108 and Acetyl-CoA Carboxylase Ser79. CONCLUSION: In a single paraffin block BHP preserved the phosphorylation state of several signaling proteins at a level comparable to snap-freezing, while maintaining the full diagnostic immunohistochemical and histomorphologic detail of formalin fixation. This new tissue fixative has the potential to greatly facilitate personalized medicine, biobanking, and phospho-proteomic research

The Importance of Context in Managerial Work: The Case of Senior Hotel Managers in Greece

Qualitative research on the interplay between context and hospitality managerial work has not been previously addressed in the literature. Based on the works of Johns (2006) and Dierdorff et al. (2009), this research suggests that luxury senior hotel managers (GMs and department managers) are to a certain degree recipients of contextual/cultural influences, depending on the ownership status of the hotel. It is also argued that managerial work is shaped and exercised by a set of moderating factors, which are unique for each individual. Overall, the aim of this research is to increase the level of awareness and trigger further research in this topic area. © 2014 Copyright © Taylor & Francis Group, LLC

Comparison of techniques for the assessment of polymorphonuclear leukocyte polarisation in suspension

Polarisation of polymorphonuclear leukocytes (PMN) is suspension was assessed using three techniques: 1) visual classification; 2) computerized morphometry; and 3) flow cytometry. While visual classification detected the formation, polarisation and type of cytoplasmic extensions produced by PMN, morphometry and flow cytometry detected only the formation of extensions. The area, perimeter and ellipticity were, in general, statistically different for each subtype of PMN-shape identified by visual classification. Furthermore, the magnitude and direction of changes detected by flow cytometry were affected by the use of erythrocyte lysis (during isolation of the cells) and the fixative used prior to analysis. The findings of this study demonstrate that visual classification is a more sensitive, reliable and appropriate assay of PMN polarisation than current morphometric and flow cytometric methods.</p

Accurate Position Monitoring and Improved Supine-Dependent Obstructive Sleep Apnea with a New Position Recording and Supine Avoidance Device

Study Objectives: Approximately 30% of obstructive sleep apnea (OSA) patients have supine-predominant OSA, and simply avoiding supine sleep should normalise respiratory disturbance event rates. However, traditional supine-avoidance therapies are inherently uncomfortable, and treatment adherence is poor and difficult to monitor objectively. This study evaluated the efficacy of a novel, potentially more acceptable position monitor and supine-avoidance device for managing supine-predominant OSA and snoring. Design and Setting: In-laboratory evaluation of position recording accuracy versus video recordings (validation study), and randomized controlled crossover trial of active versus inactive supine-avoidance therapy in the home setting (efficacy study). Patients: 17 patients undergoing in-laboratory sleep studies (validation) and 15 patients with supine-predominant OSA (efficacy). Interventions: Efficacy study: 1 week of inactive and 1 week of active treatment in randomized order, separated by 1 week. Measurements and Results: Agreement between 30-sec epoch-based posture classifications from device versus video records was high (median κ 0.95, interquartile range: 0.88-1.00), and there was good supine time agreement (bias 0.3%, 95%CI: −4.0% to 4.6%). In the efficacy study, apnea-hypopnea index (AHI) and snoring frequency were measured in-home using a nasal pressure and microphone based system during inactive and active treatment weeks. The position monitoring and supine alarm device markedly inhibited supine time (mean ± SEM 19.3% ± 4.3% to 0.4% ± 0.3%, p < 0.001) and reduced AHI (25.0 ± 1.7 to 13.7 ± 1.1 events/h, p = 0.030) but not snoring frequency. Conclusions: This new position monitoring and supine alarm device records sleep position accurately and improves OSA but not snoring in patients with supine-predominant OSA.James J. Bignold, Jeremy D. Mercer, Nick A. Antic, R. Doug McEvoy, Peter G. Catchesid

Poor Long-Term Patient Compliance with the Tennis Ball Technique for Treating Positional Obstructive Sleep Apnea

Study Objectives: Little is known regarding long-term patient compliance with the tennis ball technique (TBT), one of the original simple methods of positional therapy (i.e., avoiding the supine posture during sleep) for posture-dependent obstructive sleep apnea patients. The purpose of this study was to investigate long-term patient compliance with TBT. Methods: A follow-up questionnaire was mailed to all patients prescribed TBT at the Adelaide Institute for Sleep Health between July 2004 and March 2008 (n = 108). Results: Sixty-seven patients replied to the questionnaire. Baseline demographic/clinical characteristics were not significantly different from non-respondents. Among the respondents, follow-up time was (mean ± SD) 2.5 ± 1.0 years. Four (6.0%) reported they were still using TBT (group A); 9 (13.4%) were no longer using TBT, claiming to have learned to avoid the supine position during sleep (group B); and 54 (80.6%) were neither using TBT nor avoiding the supine posture (group C). The main reason for ceasing TBT use in group C was that TBT was too uncomfortable (34/54 patients). Conclusions: Long-term patient compliance with TBT appears to be very poor, with less than 10% of patients reporting continued use (group A) ~30 months after prescription. With most TBT non-compliers reporting it to be too uncomfortable, alternative forms of positional therapy appear to be needed.James J. Bignold, Georgina Deans-Costi, Mitchell R. Goldsworthy, Claire A. Robertson, Douglas McEvoy, Peter G. Catcheside and Jeremy D. Mercerhttp://www.aasmnet.org/JCSM/ViewAbstract.aspx?pid=2759

A study to explore the role of the community oncology nurse specialist

Cancer patients are increasingly being cared for in the community. However, the role of the community oncology nurse specialist (CONS) remains a rare appointment in the United Kingdom. There is little research evidence to substantiate the view that this is a worthwhile and effective role. This study aimed to explore the role of one CONS using a qualitative case study approach. The sample consisted of the CONS, seven patients, no relatives and five community nurses. Semi-structured interviews were used to elicit the view of these participants. In addition, documents containing details of the CONS's work to date were reviewed. A number of the CONS's role components were identified by the different respondent groups. Five themes were identified within the data collected: functions of the CONS, communication between the CONS and other health care professionals, structural and organizational factors, characteristics of the service and benefits to patients and their families. Furthermore, all respondent groups were positive about the service offered by the CONS, and felt that the role was valuable. Some negative features were identified and these were concerned with organizational aspects of the service. The findings suggest the need for further research and evaluation in this area