GliaMorph: A modular image analysis toolkit to quantify Müller glial cell morphology

Cell morphology is critical for all cell functions. This is particularly true for glial cells as they rely on complex shape to contact and support neurons. However, methods to quantify complex glial cell shape accurately and reproducibly are lacking. To address this, we developed the image analysis pipeline "GliaMorph". GliaMorph is a modular analysis toolkit developed to perform (i) image pre-processing, (ii) semi-automatic region-of-interest (ROI) selection, (iii) apicobasal texture analysis, (iv) glia segmentation, and (v) cell feature quantification. Müller Glia (MG) have a stereotypic shape linked to their maturation and physiological status. We here characterized MG on three levels, including (a) global image-level, (b) apicobasal texture, and (c) regional apicobasal vertical-to-horizontal alignment. Using GliaMorph we quantified MG development on a global and single-cell level, showing increased feature elaboration and subcellular morphological rearrangement in the zebrafish retina. As proof-of-principle, we analysed expression changes in a mouse glaucoma model, identifying subcellular protein localization changes in MG. Together, GliaMorph enables an in-depth understanding of MG morphology in the developing and diseased retina

Patient preferences and willingness-to-pay for a home or clinic based program of chronic heart failure management: findings from the which? trial

BACKGROUND Beyond examining their overall cost-effectiveness and mechanisms of effect, it is important to understand patient preferences for the delivery of different modes of chronic heart failure management programs (CHF-MPs). We elicited patient preferences around the characteristics and willingness-to-pay (WTP) for a clinic or home-based CHF-MP. METHODOLOGY/PRINCIPAL FINDINGS A Discrete Choice Experiment was completed by a sub-set of patients (n = 91) enrolled in the WHICH? trial comparing home versus clinic-based CHF-MP. Participants provided 5 choices between hypothetical clinic and home-based programs varying by frequency of nurse consultations, nurse continuity, patient costs, and availability of telephone or education support. Participants (aged 71±13 yrs, 72.5% male, 25.3% NYHA class III/IV) displayed two distinct preference classes. A latent class model of the choice data indicated 56% of participants preferred clinic delivery, access to group CHF education classes, and lower cost programs (p<0.05). The remainder preferred home-based CHF-MPs, monthly rather than weekly visits, and access to a phone advice service (p<0.05). Continuity of nurse contact was consistently important. No significant association was observed between program preference and participant allocation in the parent trial. WTP was estimated from the model and a dichotomous bidding technique. For those preferring clinic, estimated WTP was ≈AU$9-20 per visit; however for those preferring home-based programs, WTP varied widely (AU$15-105). CONCLUSIONS/SIGNIFICANCE Patient preferences for CHF-MPs were dichotomised between a home-based model which is more likely to suit older patients, those who live alone, and those with a lower household income; and a clinic-based model which is more likely to suit those who are more socially active and wealthier. To optimise the delivery of CHF-MPs, health care services should consider their patients’ preferences when designing CHF-MPs.Jennifer A. Whitty, Simon Stewart, Melinda J. Carrington, Alicia Calderone, Thomas Marwick, John D. Horowitz, Henry Krum, Patricia M. Davidson, Peter S. Macdonald, Christopher Reid, Paul A. Scuffha

Tracking the international spread of SARS-CoV-2 lineages B.1.1.7 and B.1.351/501Y-V2

Publisher Copyright: © 2021 O'Toole Á et al.Late in 2020, two genetically-distinct clusters of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) with mutations of biological concern were reported, one in the United Kingdom and one in South Africa. Using a combination of data from routine surveillance, genomic sequencing and international travel we track the international dispersal of lineages B.1.1.7 and B.1.351 (variant 501Y-V2). We account for potential biases in genomic surveillance efforts by including passenger volumes from location of where the lineage was first reported, London and South Africa respectively. Using the software tool grinch (global report investigating novel coronavirus haplotypes), we track the international spread of lineages of concern with automated daily reports, Further, we have built a custom tracking website (cov-lineages.org/global_report.html) which hosts this daily report and will continue to include novel SARS-CoV-2 lineages of concern as they are detected.Peer reviewe

Identification of elusive sequence-specific promoters of RNA polymerase II polycistronic transcription in African trypanosomes

Kinetoplastids have evolved in isolation for one billion years resulting in several divergent molecular and cellular processes. One example is protein-coding genes transcribed polycistronically by a typical RNA polymerase II (RNA pol II). Transcription most likely starts at divergent Strand Switch Regions (dSSRs), long sequences between divergently oriented polycistronic transcription units (PTUs). The lack of regulation in trypanosome transcription has become the paradigm in our field. Previous work suggests that changes in chromatin structure over broad SSR regions drives unregulated and dispersed transcription initiation. We investigate such an exceptional feature in trypanosomes by first identifying RNA pol II-enriched regions using ChIP-Seq, as potential promoter sequences. The high resolution of this technique allowed us to accurately determine peaks of RNA pol II accumulation in the dSSRs. To functionally investigate pol II-enriched sequences unbiasedly, the peaks on chromosome VII were assayed for their ability to direct transcription using transient transfection. This analysis suggests that two unidirectional short sequence specific promoters within each dSSR make up the general structure. Primer extension analysis of nascent RNA allowed us to identify precise transcription start sites (TSS) of promoters inserted in a chromosome. Detailed analysis of one of these promoters defined 75bp as sufficient to fully drive transcription and identified essential nucleotides for precise initiation around the TSS. In addition, mutations to internal and downstream boxes led to dramatic decreased activity. In summary, we show that sequence-specific unidirectional RNA pol II promoters with proper TSS transcription initiation are present in the T. brucei genome. Our results challenge the currently accepted hypothesis that trypanosomes lack true promoters with transcription initiation control

Trypanosomiasis and Leishmaniasis Symposium: Advances in Basic and Applied Research

About the British Society for Parasitology. About.Today many researchers and students are very passionate about the fascinating world of infectious diseases, parasites, their complex lifestyles and their associated impact on people and livelihoods.To draw attention to the unique importance of parasitology as a distinct discipline within biology, The British Society for Parasitology was formed in 1962 from the Parasitological Section of the Institute of Biology. Today the Society is the central networking and meeting point for many professional and amateur parasitologists throughout the UK and across the world.Did you know that the UK leads Europe – and Europe leads the world – in parasitology research? No European country publishes more parasitology research than the UK, and UK papers were cited more than those from any other country in the past 5 years (2011-2016, data from Elsevier SciVal, see News item for more details.)As the leading academic society for a country preeminent in parasitology, the remit of the BSP is broad. It promotes and supports the academic study of parasitology in all its many guises. This can be from experimental to theoretical approaches as applied to infection biology and disease research, or from ecological to medical and veterinary studies in global health and international aid. Each year students are given financial support to attend BSP meetings and scholarship schemes are in place to support fieldwork and training events.The membership of the BSP stands at around 1000 in number. Approximately a third of members are from overseas locations. Highlights of the annual BSP calendar include the annual residential meetings in spring and autumn which are focused upon general and specialist aspects of parasitology. The BSP has a close relationship with Cambridge University Press that prints a special issue on our autumn meeting.The BSP Society is a Charitable Incorporated Organisation and managed by the BSP Council. This comprises a President, Honorary Officers and ordinary Council Members, who together act as Company Trustees. Co-opted members of Council also include representation from the student, early career membership and other learned societies where clear synergies are apparent

Sample choice set.

<p>Sample choice set.</p

Latent class model coefficients.

<p>AIC Akaike Information Criterion; LL Log Likelihood.</p>***<p>p&lt;0.01.</p>**<p>p&lt;0.05.</p>*<p>p&lt;0.1.</p

Attributes and levels.

<p>Attributes and levels.</p