Publications by authors named "K R Pilkington"

50 Publications

Panel Optimization for High-Dimensional Immunophenotyping Assays Using Full-Spectrum Flow Cytometry.

Curr Protoc 2021 Sep;1(9):e222

Malaghan Institute of Medical Research, Wellington, New Zealand.

Technological advancements in fluorescence flow cytometry and an ever-expanding understanding of the complexity of the immune system have led to the development of large flow cytometry panels reaching up to 43 colors at the single-cell level. However, as panel size and complexity increase, so too does the detail involved in designing and optimizing successful high-quality panels fit for downstream high-dimensional data analysis. In contrast to conventional flow cytometers, full-spectrum flow cytometers measure the entire emission spectrum of each fluorophore across all lasers. This allows for fluorophores with very similar emission maxima but unique overall spectral fingerprints to be used in conjunction, enabling relatively straightforward design of larger panels. Although a protocol for best practices in full-spectrum flow cytometry panel design has been published, there is still a knowledge gap in going from the theoretically designed panel to the necessary steps required for panel optimization. Here, we aim to guide users through the theory of optimizing a high-dimensional full-spectrum flow cytometry panel for immunophenotyping using comprehensive step-by-step protocols. These protocols can also be used to troubleshoot panels when issues arise. A practical application of this approach is exemplified with a 24-color panel designed for identification of conventional T-cell subsets in human peripheral blood. © 2021 Malaghan Institute of Medical Research, Cytek Biosciences. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation and evaluation of optimal spectral reference controls Support Protocol 1: Antibody titration Support Protocol 2: Changing instrument settings Basic Protocol 2: Unmixing evaluation of fully stained sample Basic Protocol 3: Evaluation of marker resolution Support Protocol 3: Managing heterogeneous autofluorescence Basic Protocol 4: Assessment of data quality using expert gating and dimensionality reduction algorithms.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1002/cpz1.222DOI Listing
September 2021

The role of Kingella kingae in pre-school aged children with bone and joint infections.

J Infect 2021 09 12;83(3):321-331. Epub 2021 Jul 12.

Department of Paediatrics, University of Otago, Christchurch School of Medicine, New Zealand. Electronic address:

Objectives: The Pre-school Osteoarticular Infection (POI) study aimed to describe the burden of disease, epidemiology, microbiology and treatment of acute osteoarticular infections (OAI) and the role of Kingella kingae in these infections.

Methods: Information about children 3-60 months of age who were hospitalized with an OAI to 11 different hospitals across Australia and New Zealand between January 2012 and December 2016 was collected retrospectively.

Results: A total of 907 cases (73%) were included. Blood cultures grew a likely pathogen in only 18% (140/781). The peak age of presentation was 12 to 24 months (466/907, 51%) and Kingella kingae was the most frequently detected microorganism in this age group (60/466, 13%). In the majority of cases, no microorganism was detected (517/907, 57%). Addition of PCR to culture increased detection rates of K. kingae. However, PCR was performed infrequently (63/907, 7%).

Conclusions: This large multi-national study highlights the need for more widespread use of molecular diagnostic techniques for accurate microbiological diagnosis of OAI in pre-school aged children. The data from this study supports the hypothesis that a substantial proportion of pre-school aged children with OAI and no organism identified may in fact have undiagnosed K. kingae infection. Improved detection of Kingella cases is likely to reduce the average length of antimicrobial treatment.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.jinf.2021.06.028DOI Listing
September 2021

Towards More Predictive, Physiological and Animal-free Models: Advances in Cell and Tissue Culture 2020 Conference Proceedings.

Altern Lab Anim 2021 May 6;49(3):93-110. Epub 2021 Jul 6.

UPM-Kymmene Corporation, Helsinki, Finland.

Experimental systems that faithfully replicate human physiology at cellular, tissue and organ level are crucial to the development of efficacious and safe therapies with high success rates and low cost. The development of such systems is challenging and requires skills, expertise and inputs from a diverse range of experts, such as biologists, physicists, engineers, clinicians and regulatory bodies. Kirkstall Limited, a biotechnology company based in York, UK, organised the annual conference, (ACTC), which brought together people having a variety of expertise and interests, to present and discuss the latest developments in the field of cell and tissue culture and modelling. The conference has also been influential in engaging animal welfare organisations in the promotion of research, collaborative projects and funding opportunities. This report describes the proceedings of the latest ACTC conference, which was held virtually on 30th September and 1st October 2020, and included sessions on models in the following areas: advanced skin and respiratory models, neurological disease, cancer research, advanced models including 3-D, fluid flow and co-cultures, diabetes and other age-related disorders, and animal-free research. The roundtable session on the second day was very interactive and drew huge interest, with intriguing discussion taking place among all participants on the theme of replacement of animal models of disease.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1177/02611929211025006DOI Listing
May 2021

Evidence on yoga for health: A bibliometric analysis of systematic reviews.

Complement Ther Med 2021 Aug 4;60:102746. Epub 2021 Jun 4.

School of Health and Care Professions, University of Portsmouth, UK.

Objective: To support the research agenda in yoga for health by comprehensively identifying systematic reviews of yoga for health outcomes and conducting a bibliometric analysis to describe their publication characteristics and topic coverage.

Methods: We searched 7 databases (MEDLINE/PubMed, Embase, PsycINFO, CINAHL, AMED, the Cochrane Database of Systematic Reviews, and PROSPERO) from their inception to November 2019 and 1 database (INDMED) from inception to January 2017. Two authors independently screened each record for inclusion and one author extracted publication characteristics and topics of included reviews.

Results: We retrieved 2710 records and included 322 systematic reviews. 157 reviews were exclusively on yoga, and 165 were on yoga as one of a larger class of interventions (e.g., exercise). Most reviews were published in 2012 or later (260/322; 81 %). First/corresponding authors were from 32 different countries; three-quarters were from the USA, Germany, China, Australia, the UK or Canada (240/322; 75 %). Reviews were most frequently published in speciality journals (161/322; 50 %) complementary medicine journals (66/322; 20 %) or systematic review journals (59/322; 18 %). Almost all were present in MEDLINE (296/322; 92 %). Reviews were most often funded by government or non-profits (134/322; 42 %), unfunded (74/322; 23 %), or not explicit about funding (111/322; 34 %). Common health topics were psychiatric/cognitive (n = 56), cancer (n = 39) and musculoskeletal conditions (n = 36). Multiple reviews covered similar topics, particularly depression/anxiety (n = 18), breast cancer (n = 21), and low back pain (n = 16).

Conclusions: Further research should explore the overall quality of reporting and conduct of systematic reviews of yoga, the direction and certainty of specific conclusions, and duplication or gaps in review coverage of topics.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.ctim.2021.102746DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8350934PMC
August 2021

A relational analysis of an invisible illness: A meta-ethnography of people with chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) and their support needs.

Soc Sci Med 2020 11 16;265:113369. Epub 2020 Sep 16.

School of Social Sciences, University of Westminster, London, UK.

Chronic fatigue syndrome (CFS)/myalgic encephalomyelitis (ME) is indicated by prolonged, medically unexplained fatigue (amongst other symptoms), not alleviated by rest, and causing substantial disability. There are limited treatments on offer, which may not be effective and/or acceptable for all people, and treatment views are polarised. We, thus, aimed to take a step back from this debate, to explore more broadly preferences for formal and informal support among people with CFS/ME. We used a meta-ethnography approach to examine the substantial qualitative literature available. Using the process outlined by Noblit and Hare, and guided by patient involvement throughout, 47 studies were analysed. Our synthesis suggested that to understand people with CFS/ME (such as their invisibility, loss of self, and fraught clinical encounters), it was useful to shift focus to a 'relational goods' framework. Emotions and tensions encountered in CFS/ME care and support only emerge via 'sui generis' real life interactions, influenced by how social networks and health consultations unfold, as well as structures like disability support. This relational paradigm reveals the hidden forces at work producing the specific problems of CFS/ME, and offers a 'no blame' framework going forward.
View Article and Find Full Text PDF

Download full-text PDF

Source
http://dx.doi.org/10.1016/j.socscimed.2020.113369DOI Listing
November 2020
-->