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Accessible luminal interface of bovine rectal organoids generated from cryopreserved biopsy tissues

by Minae Kawasaki, Yoko M. Ambrosini

Developing precise species-specific in vitro models that closely resemble in vivo intestinal tissues is essential for advancing our understanding of gastrointestinal physiology and associated diseases. This is especially crucial in examining host-pathogen interactions, particularly in bovines, a known reservoir for microbes and pathogens posing substantial public health threats. This research investigated the viability of producing bovine rectal organoids from cryopreserved tissues. We compared two cryopreservation methods with a traditional technique using fresh tissues, evaluating their effectiveness through growth rates, long-term viability, and comprehensive structural, cellular, and genetic analyses. These assessments utilized phase-contrast imaging, immunofluorescence imaging, and RT-qPCR assays. Additionally, the study developed a sophisticated method for forming a functional epithelial barrier from organoid-derived bovine rectal monolayers, incorporating a wide range of epithelial cells. This methodology employed transepithelial electrical resistance (TEER), parallel artificial membrane permeability assay (Papp), confocal microscopy, and advanced imaging techniques like scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Our findings decisively show that bovine rectal organoids can be effectively generated from cryopreserved biopsy tissues. Moreover, we formulated a robust and optimized protocol for creating functional rectal monolayers from these organoids. This significant progress is particularly relevant given the susceptibility of the bovine rectum to various enteric pathogens of public health concern, marking a vital step forward in veterinary and biomedical research. The creation of accurate species specific in vitro models that faithfully mimic in vivo intestinal tissues is critical for enhancing our understanding of gut physiology and related pathologies. This is particularly relevant in studying the interactions between hosts and microbes or pathogens with significant public health risks where bovine can be the major reservoir.

Psychometric properties of the living with long term conditions scale in an English-speaking population living with long term conditions in the UK

Por: Ambrosio · L. · Hislop-Lennie · K. · Serrano-Fuentes · N. · Driessens · C. · Portillo · M. C.
Objective

To present the psychometric properties of the living with long-term condition (LwLTCs) scale in an English-speaking population of people with different LTCs.

Design

An observational and cross-sectional study, with retest was conducted. Psychometric properties including feasibility, internal consistency, confirmatory factor analysis, reproducibility and content validity were tested.

Setting

The study took place across the UK via primary care surgeries and voluntary organisations, between December 2021 and June 2022.

Participants

The study included 577 patients living with different LTCs, as chronic obstructive pulmonary disease, arthritis, chronic heart failure, Parkinson’s disease, chronic kidney disease and type 2 diabetes mellitus. Inclusion criteria included: (a) having been diagnosed with one or more of the conditions; (b) being able to read, understand and answer written questionnaires; (c) being fluent in English and (d) being able to provide written informed consent. Patients were involved in the design and pilot study of the scale.

Results

A total sample of 577 people with an age range of 37–97 years (98±9.65) were recruited. Internal consistency of the total 26-item LwLTCs scale score was excellent (ordinal alpha=0.90) but confirmatory factor analysis showed better fit indices (Normed Fit Index=0.96; standardised root mean square residual=0.051; Goodness of Fit Index=0.98) for a 20-item LwLTCs scale.

Conclusions

A shorter version of the LwLTCs scale, with just 20 items and with excellent psychometric properties, is recommended. Having a short scale is key when considering the implementation of the scale in clinical practice to develop person-centred pathways and more comprehensive care plans.

Qualitative study of COVID-19 patient experiences with non-invasive ventilation and pronation: strategies to enhance treatment adherence

Por: Brugnolli · A. · Chini · G. · Scartezzini · R. · Ambrosi · E.
Introduction

Non-invasive ventilation (NIV) treatment combined with pronation in patients with COVID-19 respiratory failure has been shown to be effective in improving respiratory function and better patient outcomes. These patients may experience discomfort or anxiety that may reduce adherence to treatment.

Objective

The aim of this study was to explore and describe the subjective experiences of patients undergoing helmet NIV and pronation during hospitalisation for COVID-19 respiratory failure, with a focus on the elements of care and strategies adopted by patients that enabled good adaptation to treatments.

Method

A qualitative descriptive study, using face-to-face interviews, was carried out with a purposeful sample of 20 participants discharged from a pulmonary intensive care unit who underwent helmet continuous positive airway pressure and pronation during hospitalisation for COVID-19.

Results

Content analysis of the transcripts revealed feelings and experiences related to illness and treatments, strategies for managing one’s own negative thoughts, and practical strategies of one’s own and healthcare workers to facilitate adaptation to pronation and helmet. Experience was reflected in five major topics related to specific time points and settings: feelings and experiences, helmet and pronation: heavy but beneficial, positive thinking strategies, patients’ practical strategies, support of healthcare professionals (HCPs).

Conclusions

This study may be useful to HCPs to improve the quality and appropriateness of care they provide.

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