Evaluation of a multi-marker immunomagnetic enrichment assay for the quantification of circulating melanoma cells

Background: Circulating melanoma cells (CMCs) are thought to be valuable in improving measures of prognosis in melanoma patients and may be a useful marker of residual disease to identify non-metastatic patients requiring adjuvant therapy. We investigated whether immunomagnetic enrichment targeting multiple markers allows more efficient enrichment of CMCs from patient peripheral blood than targeting a single marker. Furthermore, we aimed to determine whether the number of CMCs in patient blood was associated with disease stage.Methods: We captured CMCs by targeting the melanoma associated markers MCSP and MCAM as well as the melanoma stem cell markers ABCB5 and CD271, both individually and in combination, by immunomagnetic enrichment. CMCs were enriched and quantified from the peripheral blood of 10 non-metastatic and 13 metastatic melanoma patients.Results: Targeting all markers in combination resulted in the enrichment of more CMCs than when any individual marker was targeted (p \u3c 0.001-0.028). Furthermore, when a combination of markers was targeted, a greater number of CMCs were enriched in metastatic patients compared with non-metastatic patients (p = 0.007).Conclusions: Our results demonstrated that a combination of markers should be targeted for optimal isolation of CMCs. In addition, there are significantly more CMCs in metastatic patients compared with non-metastatic patients and therefore quantification of CMCs may prove to be a useful marker of disease progression

Evaluation of a multi-marker immunomagnetic enrichment assay for the quantification of circulating melanoma cells

Background: Circulating melanoma cells (CMCs) are thought to be valuable in improving measures of prognosis in melanoma patients and may be a useful marker of residual disease to identify non-metastatic patients requiring adjuvant therapy. We investigated whether immunomagnetic enrichment targeting multiple markers allows more efficient enrichment of CMCs from patient peripheral blood than targeting a single marker. Furthermore, we aimed to determine whether the number of CMCs in patient blood was associated with disease stage.Methods: We captured CMCs by targeting the melanoma associated markers MCSP and MCAM as well as the melanoma stem cell markers ABCB5 and CD271, both individually and in combination, by immunomagnetic enrichment. CMCs were enriched and quantified from the peripheral blood of 10 non-metastatic and 13 metastatic melanoma patients.Results: Targeting all markers in combination resulted in the enrichment of more CMCs than when any individual marker was targeted (p \u3c 0.001-0.028). Furthermore, when a combination of markers was targeted, a greater number of CMCs were enriched in metastatic patients compared with non-metastatic patients (p = 0.007).Conclusions: Our results demonstrated that a combination of markers should be targeted for optimal isolation of CMCs. In addition, there are significantly more CMCs in metastatic patients compared with non-metastatic patients and therefore quantification of CMCs may prove to be a useful marker of disease progression

Coating formulation change leads to inferior performance of long-lasting insecticidal nets in Papua New Guinea

Background: Long-lasting insecticidal nets (LLINs) play a key role in reducing malaria transmission in endemic countries. In a previous study, the authors demonstrated a substantial decrease in the bioefficacy of LLINs for malaria prevention delivered to Papua New Guinea (PNG) between 2013 and 2019. This coincided with a rise in malaria cases in the country. The present study was aimed at determining the underlying cause of the reduced bioefficacy observed in these LLINs. The main hypothesis was that a change in the coating formulation of the respective LLIN product was responsible, and had led to significantly altered product properties and performance. Methods: A set of PermaNet® 2.0 LLIN samples (n = 12) manufactured between 2007 and 2019 was subjected to combustion ion chromatography in order to understand the chemistry of the LLIN polymer coating formulation. In addition, World Health Organization (WHO) LLIN standard wash tests and cone bioassays were conducted to further characterize the change in product performance that occurred between 2012 and 2013. Results: High polymer fluorine content (average 3.2 g/kg) was measured in PermaNet® 2.0 manufactured up to 2012, whereas nets which were manufactured after 2012 contained very little polymer fluorine (average 0.04 g/kg) indicating a coating formulation change from a fluorocarbon (FC)-based to a non-FC-based formulation. The coating formulation change as part of the manufacturing process thus resulted in a significant reduction in bioefficacy. In addition, the manufacturing change affected wash resistance leading to a faster reduction in 24 h mosquito mortality in the non-FC-coated product with consecutive washes. Conclusion: A change in coating formulation of PermaNet® 2.0 resulted in reduced product performance in PNG. Post-2012 PermaNet® 2.0 LLINs should not be considered to be the same product as PermaNet® 2.0 LLINs produced prior to and in 2012. Coating formulation changes should be validated to not impact LLIN product performance

Diversity Exiting the Academy: Influential Factors for the Career Choice of Well-Represented and Underrepresented Minority Scientists

A national sample of PhD-trained scientists completed training, accepted subsequent employment in academic and nonacademic positions, and were queried about their previous graduate training and current employment. Respondents indicated factors contributing to their employment decision (e.g., working conditions, salary, job security). The data indicate the relative importance of deciding factors influencing career choice, controlling for gender, initial interest in faculty careers, and number of postgraduate publications. Among both well-represented (WR; n = 3444) and underrepresented minority (URM; n = 225) respondents, faculty career choice was positively associated with desire for autonomy and partner opportunity and negatively associated with desire for leadership opportunity. Differences between groups in reasons endorsed included: variety, prestige, salary, family influence, and faculty advisor influence. Furthermore, endorsement of faculty advisor or other mentor influence and family or peer influence were surprisingly rare across groups, suggesting that formal and informal support networks could provide a missed opportunity to provide support for trainees who want to stay in faculty career paths. Reasons requiring alteration of misperceptions (e.g., limited leadership opportunity for faculty) must be distinguished from reasons requiring removal of actual barriers. Further investigation into factors that affect PhDs’ career decisions can help elucidate why URM candidates are disproportionately exiting the academy

Outerwear—inner musings: A theoretical framework application for creative scholarship

Scholarship in design, namely clothing and textiles, is based in practice and research. Dire concerns and consequences face academics deciding to undertake creative production as a scholarship track, namely promotion and tenure. With a profession centered on creative practice, efforts to better define and document methodological rigor for creative scholarship must be made, in order to increase knowledge dissemination across the discipline. Studies in information systems design-science theory application indicate various guidelines for undertaking creative practice as design-science (Hevner et al., 2004)

Teaching design research through practice: a pilot study for collaborative exploration

As design educators, we experience tension between devoting the precious little time we have in class to educating our students in ways of making (i.e., skills in sewing and patternmaking) and ways of thinking (i.e. design ideation, creativity, etc.)

Temporal changes in innate immune signals in a rat model of alcohol withdrawal in emotional and cardiorespiratory homeostatic nuclei.

BACKGROUND: Chronic alcohol use changes the brain\u27s inflammatory state. However, there is little work examining the progression of the cytokine response during alcohol withdrawal, a period of profound autonomic and emotional upset. This study examines the inflammatory response in the central nucleus of the amygdala (CeA) and dorsal vagal complex (DVC), brain regions neuroanatomically associated with affective and cardiorespiratory regulation in an in vivo rat model of withdrawal following a single chronic exposure. METHODS: For qRT-PCR studies, we measured the expression of TNF-α, NOS-2, Ccl2 (MCP-1), MHC II invariant chain CD74, and the TNF receptor Tnfrsf1a in CeA and DVC samples from adult male rats exposed to a liquid alcohol diet for thirty-five days and in similarly treated animals at four hours and forty-eight hours following alcohol withdrawal. ANOVA was used to identify statistically significant treatment effects. Immunohistochemistry (IHC) and confocal microscopy were performed in a second set of animals during chronic alcohol exposure and subsequent 48-hour withdrawal. RESULTS: Following a chronic alcohol exposure, withdrawal resulted in a statistically significant increase in the expression of mRNAs specific for innate immune markers Ccl2, TNF-α, NOS-2, Tnfrsf1a, and CD74. This response was present in both the CeA and DVC and most prominent at 48 hours. Confocal IHC of samples taken 48 hours into withdrawal demonstrate the presence of TNF-α staining surrounding cells expressing the neural marker NeuN and endothelial cells colabeled with ICAM-1 (CD54) and RECA-1, markers associated with an inflammatory response. Again, findings were consistent in both brain regions. CONCLUSIONS: This study demonstrates the rapid induction of Ccl2, TNF-α, NOS-2, Tnfrsf1a and CD74 expression during alcohol withdrawal in both the CeA and DVC. IHC dual labeling showed an increase in TNF-α surrounding neurons and ICAM-1 on vascular endothelial cells 48 hours into withdrawal, confirming the inflammatory response at the protein level. These findings suggest that an abrupt cessation of alcohol intake leads to an acute central nervous system (CNS) inflammatory response in these regions that regulate autonomic and emotional state

The National Competency Framework for registered nurses in adult critical care: An overview

In the years following the abolition of the English National Board for Nursing, Midwifery and Health Visiting (ENB) in 2002, concerns were raised within the Critical Care nursing community about a lack of consistency in post-registration education programmes. In response to this the Critical Care Network National Nurse Leads (CC3N) formed a sub-group, the Critical Care Nurse Education Review Forum (CCNERF) to address these concerns. A review of UK course provision confirmed marked inconsistency in the length, content and associated academic award. The CCNERF commenced a two phase project, first developing national standards for critical care nurse education such as length of course and academic credit level; followed by the development of a national competency framework1, 2. Following significant review and revision, version two of the National Competency Framework for Registered Nurses in Adult Critical Care was published by CC3N in 20153. This paper introduces the National Competency Framework and provides an overview of its background, development and implementation. It then considers the future direction of UK post-registration Critical Care nurse education