Single-Cell RNA Sequencing Reveals Metabolic Stress-Dependent Activation of Cardiac Macrophages in a Model of Dyslipidemia-Induced Diastolic Dysfunction

Circulation, Ahead of Print. BACKGROUND:Metabolic distress is often associated with heart failure with preserved ejection fraction (HFpEF) and represents a therapeutic challenge. Metabolism-induced systemic inflammation links comorbidities with HFpEF. How metabolic changes affect myocardial inflammation in the context of HFpEF is not known.METHODS:We found that ApoE knockout mice fed a Western diet recapitulate many features of HFpEF. Single-cell RNA sequencing was used for expression analysis of CD45+cardiac cells to evaluate the involvement of inflammation in diastolic dysfunction. We focused bioinformatics analysis on macrophages, obtaining high-resolution identification of subsets of these cells in the heart, enabling us to study the outcomes of metabolic distress on the cardiac macrophage infiltrate and to identify a macrophage-to-cardiomyocyte regulatory axis. To test whether a clinically relevant sodium glucose cotransporter-2 inhibitor could ameliorate the cardiac immune infiltrate profile in our model, mice were randomized to receive the sodium glucose cotransporter-2 inhibitor dapagliflozin or vehicle for 8 weeks.RESULTS:ApoE knockout mice fed a Western diet presented with reduced diastolic function, reduced exercise tolerance, and increased pulmonary congestion associated with cardiac lipid overload and reduced polyunsaturated fatty acids. The main immune cell types infiltrating the heart included 4 subpopulations of resident and monocyte-derived macrophages, determining a proinflammatory profile exclusively in ApoE knockout- Western diet mice. Lipid overload had a direct effect on inflammatory gene activation in macrophages, mediated through endoplasmic reticulum stress pathways. Investigation of the macrophage-to-cardiomyocyte regulatory axis revealed the potential effects on cardiomyocytes of multiple inflammatory cytokines secreted by macrophages, affecting pathways such as hypertrophy, fibrosis, and autophagy. Finally, we describe an anti-inflammatory effect of sodium glucose cotransporter-2 inhibitor in this model.CONCLUSIONS:Using single-cell RNA sequencing , in a model of diastolic dysfunction driven by hyperlipidemia, we have determined the effects of metabolic distress on cardiac inflammatory cells, in particular on macrophages, and suggest sodium glucose cotransporter-2 inhibitors as potential therapeutic agents for the targeting of a specific phenotype of HFpEF.

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Dicembre 2023

Psychometric validation of the Perceived Stress Scale (PSS-10) among family caregivers of people with schizophrenia in China

Background
The 10-item Perceived Stress Scale (PSS-10) is a widely used measure of perceived stress that has been validated in various populations, yet with inconsistent results on its factor structure. The present study examines the reliability and validity of the PSS-10 in a population not previously examined: Chinese family caregivers of persons with schizophrenia, with a focus on factor analysis.

Methods
A sample of 449 family caregivers of persons with schizophrenia was recruited for psychometric testing of the scale. The factor structure of PSS-10 was tested by randomly dividing the sample into two groups for both exploratory factor analysis (EFA) and confirmatory factor analysis (CFA). The scale was further tested for internal consistency, test–retest reliability, convergent validity, discriminant validity and concurrent validity.

Results
EFA extracted two factors: perceived helplessness with six negative phrasing items and perceived efficacy with four positive phrasing items. CFA confirmed the structure of two factors with satisfactory model fit indices. Convergent validity was supported by high standard regression weight (0.78–0.92), average variance extracted (AVE=0.79–0.81) and composite reliability (0.88–0.94), while discriminant validity was confirmed by higher AVE estimates than the squared interconstruct correlations. The PSS-10 showed good internal consistency and test–retest reliability, with Cronbach’s alpha of 0.79 and intraclass correlation coefficient of 0.91, respectively. Concurrent validity was demonstrated by its significant positive correlations with stigma, depression and anxiety, as well as significant negative correlations with social support, family functioning and positive caregiving experiences.

Conclusion
The two-factor PSS-10 has good psychometric characteristics assessing the perceived stress of family caregivers of people with schizophrenia. The findings indicate that the PSS-10 can be used to measure perceived stress in future research and practice among caregivers of people with schizophrenia, and potentially, other caregiving samples.

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Novembre 2023

Work-related stress and associated factors among health professionals working in Ambo town public health facilities, West Shoa Zone, Ethiopia, 2021: a cross-sectional study

Objectives
To assess work-related stress (WRS) and associated factors among health professionals working in Ambo town governmental health facilities, in 2021.

Design
Institution-based analytical cross-sectional study.

Setting
Institution-based analytical cross-sectional study was conducted in Ambo town from 15 July 2021 to 15 August 2021. A simple random sampling technique was used to select 420 participants. Data were collected by using structured self-administered questionnaire. Descriptive statistics was used to show the magnitude of WRS. Multivariate logistic regression was employed to identify variables that are significantly associated with WRS at 95% CI and p value

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Novembre 2023

REVISE: Re-Evaluating the Inhibition of Stress Erosions in the ICU: a randomised trial protocol

Introduction
The Re-Evaluating the Inhibition of Stress Erosions (REVISE) Trial aims to determine the impact of the proton pump inhibitor pantoprazole compared with placebo on clinically important upper gastrointestinal (GI) bleeding in the intensive care unit (ICU), 90-day mortality and other endpoints in critically ill adults. The objective of this report is to describe the rationale, methodology, ethics and management of REVISE.

Methods and analysis
REVISE is an international, randomised, concealed, stratified, blinded parallel-group individual patient trial being conducted in ICUs in Canada, Australia, Saudi Arabia, UK, US, Kuwait, Pakistan and Brazil. Patients≥18 years old expected to remain invasively mechanically ventilated beyond the calendar day after enrolment are being randomised to either 40 mg pantoprazole intravenously or an identical placebo daily while mechanically ventilated in the ICU. The primary efficacy outcome is clinically important upper GI bleeding within 90 days of randomisation. The primary safety outcome is 90-day all-cause mortality. Secondary outcomes include rates of ventilator-associated pneumonia, Clostridioides difficile infection, new renal replacement therapy, ICU and hospital mortality, and patient-important GI bleeding. Tertiary outcomes are total red blood cells transfused, peak serum creatinine level in the ICU, and duration of mechanical ventilation, ICU and hospital stay. The sample size is 4800 patients; one interim analysis was conducted after 2400 patients had complete 90-day follow-up; the Data Monitoring Committee recommended continuing the trial.

Ethics and dissemination
All participating centres receive research ethics approval before initiation by hospital, region or country, including, but not limited to – Australia: Northern Sydney Local Health District Human Research Ethics Committee and Mater Misericordiae Ltd Human Research Ethics Committee; Brazil: Comissão Nacional de Ética em Pesquisa; Canada: Hamilton Integrated Research Ethics Board; Kuwait: Ministry of Health Standing Committee for Coordination of Health and Medical Research; Pakistan: Maroof Institutional Review Board; Saudi Arabia: Ministry of National Guard Health Affairs Institutional Review Board: United Kingdom: Hampshire B Research Ethics Committee; United States: Institutional Review Board of the Nebraska Medical Centre. The results of this trial will inform clinical practice and guidelines worldwide.

Trial registration number
NCT03374800.

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Novembre 2023

Abstract 16945: Aberrant Citric Acid Cycle Metabolites Temporally Coincide With Myocardial Dysfunction in a Rat Model of Stress Cardiomyopathy

Circulation, Volume 148, Issue Suppl_1, Page A16945-A16945, November 6, 2023. Introduction:Stress cardiomyopathy is characterized by transient contractile dysfunction with a poorly understood mechanism. Targeting metabolic derangements has proven efficacious in chronic heart failure, but the role of metabolism in acute cardiac dysfunction is ill-defined.Hypothesis:Characterizing any metabolic alterations in stress cardiomyopathy may help identify potential therapeutic targets.Methods:Sprague-Dawley rats (350 ± 57 g) were given a single intraperitoneal injection of 75 mg/kg isoprenaline. Animals were allocated into groups (N=30 each) based on designated time-points for assessment after injection: control (no injection), day 1, 3, and 7. Rats underwent left ventricular PV loop catheterization followed by harvesting of apical ventricular tissue. Targeted metabolomic analyses were performed via LC-MS and supplemented with enzyme activity assays, qPCR, and Western blotting.Results:Stroke work, cardiac output, and ejection fraction were all significantly reduced by day 1 with recovery by day 7 (Figure A, all p0.05).Conclusions:Profound rises in citric acid metabolites with accompanying increases in anaplerotic amino acids are suggestive of metabolic reprogramming in the setting of energetic deficits. Notably, these trends differ from those found in chronic heart failure, and represent a novel phenotypic characterization of stress cardiomyopathy with potential therapeutic implications.

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Novembre 2023