Dataset Information

Whose Gene Is It Anyway? The Effect of Preparation Purity on Neutrophil Transcriptome Studies.

ABSTRACT: Protocols for the isolation of neutrophils from whole blood often result in neutrophil preparations containing low numbers (~5%) of contaminating leukocytes, and it is possible that these contaminating cells contribute to highly sensitive assays that measure neutrophil gene expression (e.g. qPCR). We investigated the contribution of contaminating leukocytes on the transcriptome profile of human neutrophils following stimulation with inflammatory cytokines (GM-CSF, TNF?), using RNA-Seq. Neutrophils were isolated using Polymorphprep or the StemCell untouched neutrophil isolation kit (negative selection of "highly pure" neutrophils). The level of contamination was assessed by morphology and flow cytometry. The major source of contamination in Polymorphprep neutrophil preparations was from eosinophils and was highly donor dependent. Contaminating cells were largely, but not completely, absent in neutrophil suspensions prepared using negative selection, but the overall yield of neutrophils was decreased by around 50%. RNA-seq analysis identified only 25 genes that were significantly differentially-expressed between Polymorphprep and negatively-selected neutrophils across all three treatment groups (untreated, GM-CSF, TNF?). The expression levels of 34 cytokines/chemokines both before and after GM-CSF or TNF? treatment were not significantly different between neutrophil isolation methods and therefore not affected by contributions from non-neutrophil cell types. This work demonstrates that low numbers (<5%) of contaminating leukocytes in neutrophil preparations contribute very little to the overall gene expression profile of cytokine-stimulated neutrophils, and that protocols for the isolation of highly pure neutrophils result in significantly lower yields of cells which may hinder investigations where large numbers of cells are required or where volumes of blood are limited.

SUBMITTER: Thomas HB

PROVIDER: S-EPMC4581699 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Whose Gene Is It Anyway? The Effect of Preparation Purity on Neutrophil Transcriptome Studies.

Thomas Huw B HB Moots Robert J RJ Edwards Steven W SW Wright Helen L HL

PloS one 20150924 9

Protocols for the isolation of neutrophils from whole blood often result in neutrophil preparations containing low numbers (~5%) of contaminating leukocytes, and it is possible that these contaminating cells contribute to highly sensitive assays that measure neutrophil gene expression (e.g. qPCR). We investigated the contribution of contaminating leukocytes on the transcriptome profile of human neutrophils following stimulation with inflammatory cytokines (GM-CSF, TNFα), using RNA-Seq. Neutrophi ...[more]

PMID: 26401909

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Project description:AIM:The aims of this literature review were to better understand the current literature about person-centred care (PCC) and identify a clear definition of the term PCC relevant to nursing practice. METHOD/DATA SOURCES:An integrative literature review was undertaken using The Cumulative Index to Nursing and Allied Health Literature (CINAHL), Medline, Scopus and Pubmed databases. The limitations were English language, full text articles published between 1998 and 2018 within Australian, New Zealand, Canada, USA, Europe, Ireland and UK were included. The international context off PCC is then specifically related to the Australian context. REVIEW METHODS:The review adopted a thematic analysis to categorise and summarise themes with reference to the concept of PCC. The review process also adhered to the Preferred Reporting System for Meta-Analysis (PRISMA) and applied the Critical Appraisal Skills Programme (CASP) tools to ensure the quality of the papers included for deeper analysis. RESULTS:While definitions of PCC do exist, there is no universally used definition within the nursing profession. This review has found three core themes which contribute to how PCC is understood and practiced, these are People, Practice and Power. This review uncovered a malalignment between the concept of PCC and the operationalisation of the term; this misalignment was discovered at both the practice level, and at the micro, meso and micro levels of the healthcare service. CONCLUSION:The concept of PCC is well known to nurses, yet ill-defined and operationalised into practice. PCC is potentially hindered by its apparent rhetorical nature, and further investigation of how PCC is valued and operationalised through its measurement and reported outcomes is needed. Investigation of the literature found many definitions of PCC, but no one universally accepted and used definition. Subsequently, PCC remains conceptional in nature, leading to disparity between how it is interpreted and operationalised within the healthcare system and within nursing services.

Project description:Social psychologists have not fully investigated the role of leadership in mobilizing widespread support for social change, particularly gender equality. The burden of achieving gender equality is typically placed on women (particularly female leaders) - the main targets of such inequality. Traditional approaches frame workplace gender equality as either a women's issue, which limits men's (non-target's) involvement in the movement, or a meritocratic non-issue that exists due to women's (target's) tendency to pursue less intensive careers. In contrast to such work focusing on women's experiences as targets of discrimination or men's role in preserving inequality, we propose a solidarity-based approach that positions men and women as agents of change. This approach relies on two processes: leadership processes - particularly leadership as a form of influence based on shared identities among leaders and followers (e.g., their gender group); and political solidarity as a way to mobilize the silent majority (men) to work as allies beside a minority (women) and embrace equality as a common cause for both groups. In two experiments (Ns = 338, 336) we studied how leader gender and message framing affect men's and women's support for equality by contrasting a solidarity-based framing of gender equality as a common cause for men and women, with a women's issue frame (Experiment 1) or a meritocratic frame (Experiment 2). The statement was attributed to a male or female leader (Experiments 1-2) or, additionally, to a government agency (Experiment 1). Women reported higher sense of common cause (Experiment 2) and collective action intentions than men (Experiments 1-2), and higher intentions under common cause compared to meritocracy frames (Experiment 2). Interestingly, male leaders invoked higher sense of common cause and collective action intentions for both men and women regardless of framing (Experiment 2). Irrespective of leader gender however, as predicted common cause framing boosted perceived leader prototypicality, legitimacy, and influence across the board (Experiments 1-2). Yet this was qualified by women (compared to men) rating leaders as more legitimate and influential under common cause compared to meritocracy framing (Experiment 2). Women's reactions to equality messages, and the intersection of leadership and solidarity toward equality are discussed.

Dataset Information

Whose Gene Is It Anyway? The Effect of Preparation Purity on Neutrophil Transcriptome Studies.

Publications

Whose Gene Is It Anyway? The Effect of Preparation Purity on Neutrophil Transcriptome Studies.

Similar Datasets

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