A WNT mimetic with broad spectrum FZD-specificity decreases fibrosis and improves function in a pulmonary damage model.

BACKGROUND: Wnt/ß-catenin signaling is critical for lung development and AT2 stem cell maintenance in adults, but excessive pathway activation has been associated with pulmonary fibrosis, both in animal models and human diseases such as idiopathic pulmonary fibrosis (IPF). IPF is a detrimental interstitial lung disease, and although two approved drugs limit functional decline, transplantation is the only treatment that extends survival, highlighting the need for regenerative therapies. METHODS: Using our antibody-based platform of Wnt/ß-catenin modulators, we investigated the ability of a pathway antagonist and pathway activators to reduce pulmonary fibrosis in the acute bleomycin model, and we tested the ability of a WNT mimetic to affect alveolar organoid cultures. RESULTS: A WNT mimetic agonist with broad FZD-binding specificity (FZD1,2,5,7,8) potently expanded alveolar organoids. Upon therapeutic dosing, a broad FZD-binding specific Wnt mimetic decreased pulmonary inflammation and fibrosis and increased lung function in the bleomycin model, and it impacted multiple lung cell types in vivo. CONCLUSIONS: Our results highlight the unexpected capacity of a WNT mimetic to effect tissue repair after lung damage and support the continued development of Wnt/ß-catenin pathway modulation for the treatment of pulmonary fibrosis.

Sex differences in left-ventricular strain in a murine model of coxsackievirus B3 myocarditis.

Myocarditis is typically caused by viral infections, but most cases are thought to be subclinical. Echocardiography is often used for initial assessment of myocarditis patients but is poor at detecting subtle changes in cardiac dysfunction. Cardiac strain, such as global longitudinal strain (GLS) and global circumferential strain (GCS), represents an increasingly used set of measurements which can detect these subtle changes. Using a murine model of coxsackievirus B3 myocarditis, we characterized functional changes in the heart using echocardiography during myocarditis and by sex. We found that 2D GLS, 4D mode, and 4D strains detected a significant reduction in ejection fraction and GLS during myocarditis compared to baseline and in males compared to females. Furthermore, worse GLS correlated to increased levels of CD45+, CD11b+, and CD3+ immune cells. Our findings closely resemble published reports of GLS in patients with myocarditis indicating the usefulness of this animal model for translational studies of myocarditis.

Does COVID-19 impair V̇o2peak in patients with cardiorespiratory disease? Insight from cardiopulmonary responses to maximal exercise pre- and post-illness.

Reduced exercise capacity has been suggested as a cardinal sequela of COVID-19. However, only cross-sectional approaches that either do not consider individuals with concomitant cardiorespiratory disease or account for exercise capacity before infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) support this assumption. Is reduced exercise capacity a sequela of SARS-CoV-2 infection in patients with concomitant cardiorespiratory disease? We retrospectively reviewed cardiopulmonary exercise testing (CPET) data collected across three hospitals between October 2018 and March 2022. Forty-two patients who completed a CPET before and after COVID-19 and 25 patients who performed two separate CPETs but did not contract COVID-19 (CTL) were included. Within each patient, the same test protocol was performed at the first and second CPETs. The time between CPETs was similar between the groups (COVID-19 489 ± 534 vs. CTL 534 ± 257 days, P = 0.662). The COVID-19 group performed the CPETs 312 ± 232 days before and 176 ± 110 days after infection. Exercise time, peak heart rate, peak systolic pressure, oxygen uptake (V̇o2) at anaerobic threshold, peak ventilation, and ventilatory efficiency were not different between the CPETs in both groups. Peak V̇o2 was reduced from before to after SARS-CoV-2 infection. However, the change in V̇o2peak from the first to the second CPET was not different between COVID-19 vs. CTL. Accounting for V̇o2peak before COVID-19 and including a group of control patients, we find limited evidence for reduced exercise capacity as a sequela of SARS-CoV-2 infection in patients with concomitant cardiorespiratory disease.NEW & NOTEWORTHY There is accumulating evidence that reduced exercise capacity is, or can be, an outcome following COVID-19. However, evidence to date relies upon cross-sectional approaches that either do not consider patients with concomitant cardiorespiratory disease or account for pre-infection exercise capacity data. Accounting for V̇o2peak before COVID-19 and including a group of control patients, we find limited evidence for reduced exercise capacity as a sequela of SARS-CoV-2 infection in patients with concomitant cardiorespiratory disease.

Sex differences in mitochondrial gene expression during viral myocarditis.

Background: Myocarditis is an inflammation of the heart muscle most often caused by an immune response to viral infections. Sex differences in the immune response during myocarditis have been well described but upstream mechanisms in the heart that might influence sex differences in disease are not completely understood. Methods: Male and female BALB/c wild type mice received an intraperitoneal injection of heart-passaged coxsackievirus B3 (CVB3) or vehicle control. Bulk-tissue RNA-sequencing was conducted to better understand sex differences in CVB3 myocarditis. We performed enrichment analysis to understand sex differences in the transcriptional landscape of myocarditis and identify candidate transcription factors that might drive sex differences in myocarditis. Results: The hearts of male and female mice with myocarditis were significantly enriched for pathways related to an innate and adaptive immune response compared to uninfected controls. When comparing females to males with myocarditis, males were enriched for inflammatory pathways and gene changes that suggested worse mitochondrial transcriptional support (e.g., mitochondrial electron transport genes). In contrast, females were enriched for pathways related to mitochondrial respiration and bioenergetics, which were confirmed by higher transcript levels of master regulators of mitochondrial function including peroxisome proliferator-activated receptor gamma coactivator 1 (PGC1α), nuclear respiratory factor 1 (NRF1) and estrogen-related receptor alpha (ERRα). TRANSFAC analysis identified ERRa as a transcription factor that may mediate sex differences in mitochondrial function during myocarditis. Conclusions: Master regulators of mitochondrial function were elevated in females with myocarditis compared to males and may promote sex differences in mitochondrial respiratory transcript expression during viral myocarditis resulting in less severe myocarditis in females following viral infection.

Routine, ensemble characterisation of electrophoretic mobility in high and saturated ionic dispersions.

With the industrialisation of nanoparticle manufacture, the pervasive incursion of nanoparticles into the environment, the need to characterise nano-scale pharmaceuticals and living systems in replicated in vivo conditions, the continuing development of new theories to describe the electro-kinetic behaviour of nano-particles in representative ionic strengths and numerous other applications, there is an urgent requirement to provide simple and effective experimental tools to validate these models and explore new systems. Micro-electrophoresis implemented with a diffusion barrier, which isolates the dispersed phase from the electrode surface, is demonstrated as enabling such measurements for the first time, preventing the catastrophic outgassing, precipitation and sample degradation observed when the dispersed phase is in close proximity to the electrode surface. Using a measurement of a few minute's duration in a standard laboratory light scattering instrument we reproduce the theoretically predicted phenomena of asymptotic, non-zero electrophoretic mobility with increasing ionic strength, the cationic Hofmeister series dependency, charge inversion and a continuously decreasing variation in mobility with pH as molarity increases. Standard operating procedures are developed and included to encourage further work.

Reaction of O2 with a diiron protein generates a mixed-valent Fe2+/Fe3+ center and peroxide.

The gene encoding the cyanobacterial ferritin SynFtn is up-regulated in response to copper stress. Here, we show that, while SynFtn does not interact directly with copper, it is highly unusual in several ways. First, its catalytic diiron ferroxidase center is unlike those of all other characterized prokaryotic ferritins and instead resembles an animal H-chain ferritin center. Second, as demonstrated by kinetic, spectroscopic, and high-resolution X-ray crystallographic data, reaction of O2 with the di-Fe2+ center results in a direct, one-electron oxidation to a mixed-valent Fe2+/Fe3+ form. Iron-O2 chemistry of this type is currently unknown among the growing family of proteins that bind a diiron site within a four α-helical bundle in general and ferritins in particular. The mixed-valent form, which slowly oxidized to the more usual di-Fe3+ form, is an intermediate that is continually generated during mineralization. Peroxide, rather than superoxide, is shown to be the product of O2 reduction, implying that ferroxidase centers function in pairs via long-range electron transfer through the protein resulting in reduction of O2 bound at only one of the centers. We show that electron transfer is mediated by the transient formation of a radical on Tyr40, which lies ∼4 Å from the diiron center. As well as demonstrating an expansion of the iron-O2 chemistry known to occur in nature, these data are also highly relevant to the question of whether all ferritins mineralize iron via a common mechanism, providing unequivocal proof that they do not.

Completed audit cycle to explore the use of the STOPP/START toolkit to optimise medication in psychiatric in-patients with dementia.

Aims and method To explore the use of the STOPP/START toolkit in older psychiatric in-patients with dementia. Clinical records and current drug charts were reviewed against STOPP/START criteria for all in-patients (n = 86) on six specialist dementia wards. RESULTS: Benzodiazepines, antipsychotics and opiates were most commonly prescribed inappropriately. The most common unprescribed medication groups were statins, calcium supplements and vitamin D supplements. There was an overall reduction of 7% in comorbidities and 8% in the number of prescriptions. t-test showed a significant drop in average comorbidities between both audits, t(1) = 23.920, P = 0.027, and in average prescriptions per patient, t(1) = 28.808, P = 0.022. There was no difference in the number of patients receiving polypharmacy, t(1) = 7.500, P = 0.084, or receiving medication with a high risk of adverse drug reactions, t(1) = 6.857, P = 0.092. Clinical implications The STOPP/START toolkit highlighted the importance of collaborative working between doctors, clinical pharmacists and nursing staff, and could provide old age psychiatrists with a structured tool to identify inappropriate prescribing of non-psychiatric medications. Declaration of interests None.

Influence of graft size on graft survival following Descemet stripping automated endothelial keratoplasty.

PURPOSE: To evaluate graft size on outcome following Descemet stripping automated endothelial keratoplasty (DSAEK) METHODS: Consecutive patients who had undergone a DSAEK for Fuchs endothelial dystrophy (FED) and pseudophakic bullous keratopathy (PBK) with at least 1 year of follow-up. Patients were divided into three groups according to the size of the donor trephine: <9, 9 and 9.5 mm. Main outcomes were postoperative best corrected visual acuity (BCVA) and graft failure. Grafts were prepared using an automated microkeratome. For larger grafts (≥9 mm), a manual dissection of the residual peripheral ring of anterior lamella was performed before trephination. Donor age, endothelial cell density (ECD) and postmortem times; recipient details including risk factors, comorbidity, surgical complications and postoperative BCVA and graft survival were analysed. RESULTS: Of 174 patients, 131 were included: 84 (64%) with FED and 47 (36%) with PBK. Mean preoperative and postoperative BCVA were 1.01±0.76 and 0.2±0.2 logMAR, respectively, at 12 months with 80.5% achieving 20/40 or better. Postoperative BCVA was significantly associated with ECD (p=0.005), PBK or FED (p=0.004), risk factors (p=0.007) and comorbidity (p=0.016). Eleven patients (8.40%) experienced endothelial graft failure; 17.86% for <9 mm, 7.69% for 9 mm and 3.84% for 9.5 mm trephine sized grafts. Graft failure was significantly associated with ECD (p=0.039) and graft trephine size (p=0.04). CONCLUSIONS: Larger grafts occupy a smaller chord length in the eye than the trephine size and are expected to provide 10%-20% more endothelial cells. Increased graft size and donor ECD is significantly associated with a reduced graft failure rate.

Multi-marker assessment of ovarian reserve predicts oocyte yield after ovulation induction.

BACKGROUND: Many hormone and ultrasound measurements have been assessed as possible markers of ovarian reserve and to identify potential poor responders to ovulation induction. The objective of this study is to determine whether multiple biomarkers measured in blood samples collected immediately before commencement of ovulation induction for IVF can predict the outcome of ovarian stimulation. METHODS: We conducted a prospective observational study, including 356 unselected women undergoing ovulation induction/IVF at two centers. Anti-Müllerian hormone (AMH), inhibin B and FSH were measured before commencement of ovulation induction. The main outcome measures were the number of oocytes retrieved and pregnancy outcome. RESULTS: Univariate analyses showed that age, FSH, inhibin B and AMH were significant predictors for poor oocyte yield. AMH presented the highest receiver operating characteristic area under the curve (ROC(AUC)) of 0.827 indicating a good discriminating potential for predicting poor ovarian response, followed by FSH with an ROC(AUC) of 0.721. In the multivariate analysis, the variables age, FSH and AMH remained significant and the resulting model provided a high ROC(AUC) of 0.819. Women with an ovarian reserve test of <0.3 have more than a 75% chance of having their treatment cycle canceled, but a value over 0.73 indicates a 38% chance of pregnancy. Number of oocytes and oocyte yield per unit FSH administered were correlated with log model for no pregnancy (r = -0.217, P < 0.001 and r = -0.367, P < 0.001, respectively) but had limited predictive value. CONCLUSIONS: A derived estimate of ovarian reserve demonstrated superior ability for predicting oocyte yield after ovulation induction when compared with any single endocrine marker (AMH, inhibin B, FSH).