Introduction: Aging impairs the function of the central circadian clock in mammals, the suprachiasmatic nucleus (SCN), leading to a reduction in the output signal. The weaker timing signal from the... Show moreIntroduction: Aging impairs the function of the central circadian clock in mammals, the suprachiasmatic nucleus (SCN), leading to a reduction in the output signal. The weaker timing signal from the SCN results in a decline in rhythm strength in many physiological functions, including sleep–wake patterns. Accumulating evidence suggests that the reduced amplitude of the SCN signal is caused by a decreased synchrony among the SCN neurons. The present study was aimed to investigate the hypothesis that the excitation/inhibition (E/I) balance plays a role in synchronization within the network.Methods: Using calcium (Ca2+) imaging, the polarity of Ca2+ transients in response to GABA stimulation in SCN slices of old mice (20–24 months) and young controls was studied.Results: We found that the amount of GABAergic excitation was increased, and that concordantly the E/I balance was higher in SCN slices of old mice when compared to young controls. Moreover, we showed an effect of aging on the baseline intracellular Ca2+ concentration, with higher Ca2+ levels in SCN neurons of old mice, indicating an alteration in Ca2+ homeostasis in the aged SCN. We conclude that the change in GABAergic function, and possibly the Ca2+ homeostasis, in SCN neurons may contribute to the altered synchrony within the aged SCN network. Show less
A. das; Kelly, C.; Ben-Arzi, H.; Geest, R.J. van der; Plein, S.; Dall'Armellina, E. 2022
Background: Despite advancements in percutaneous coronary intervention, a significant proportion of ST-elevation myocardial infarction (STEMI) survivors develop long-term adverse left ventricular ... Show moreBackground: Despite advancements in percutaneous coronary intervention, a significant proportion of ST-elevation myocardial infarction (STEMI) survivors develop long-term adverse left ventricular (LV) remodelling, which is associated with poor prognosis. Adverse remodelling is difficult to predict, however four-dimensional (4D) flow cardiovascular magnetic resonance (CMR) can measure various aspects of LV intra-cavity flow beyond LV ejection fraction and is well equipped for exploring the underlying mechanical processes driving remodelling. The aim for this study was to compare acute 4D flow CMR parameters between patients who develop adverse remodelling with patients who do not. Methods: Fifty prospective 'first-event' STEMI patients underwent CMR 5 days post-reperfusion, which included cine-imaging, and 4D flow for assessing in-plane kinetic energy (KE), residual volume, peak-E and peak-A wave KE (indexed for LV end-diastolic volume [LVEDV]). All subjects underwent follow-up cine CMR imaging at 12 months to identify adverse remodelling (defined as 20% increase in LVEDV from baseline). Quantitative variables were compared using unpaired student's t-test. Tests were deemed statistically significant when p < 0.05. Results: Patients who developed adverse LV remodelling by 12 months had significantly higher in-plane KE (54 +/- 12 vs 42 +/- 10%, p = 0.02), decreased proportion of direct flow (27 +/- 9% vs 11 +/- 4%, p < 0.01), increased proportion of delayed ejection flow (22 +/- 9% vs 12 +/- 2, p < 0.01) and increased proportion of residual volume after 2 consecutive cardiac cycles (64 +/- 14 vs 34 +/- 14%, p < 0.01), in their acute scan. Conclusion: Following STEMI, increased in-plane KE, reduced direct flow and increased residual volume in the acute scan were all associated with adverse LV remodelling at 12 months. Our results highlight the clinical utility of acute 4D flow in prognostic stratification in patients following myocardial infarction. Show less
Ruiz, A.R.; Tuerlings, M.; A. das; Almeida, R.C. de; Suchiman, H.E.D.; Nelissen, R.G.H.H.; ... ; Meulenbelt, I. 2022
Objectives OA is a complex genetic disease with different risk factors contributing to its development. One of the genes, TNFRSF11B, previously identified with gain-of-function mutation in a family... Show moreObjectives OA is a complex genetic disease with different risk factors contributing to its development. One of the genes, TNFRSF11B, previously identified with gain-of-function mutation in a family with early-onset OA with chondrocalcinosis, is among the highest upregulated genes in lesioned OA cartilage (RAAK-study). Here, we determined the role of TNFRSF11B overexpression in development of OA. Methods Human primary articular chondrocytes (9 donors RAAK study) were transduced using lentiviral particles with or without TNFRSF11B. Cells were cultured for 1 week in a 3 D in-vitro chondrogenic model. TNFRSF11B overexpression was confirmed by RT-qPCR, immunohistochemistry and ELISA. Effects of TNFRSF11B overexpression on cartilage matrix deposition, matrix mineralization, and genes highly correlated to TNFRSF11B in RNA-sequencing dataset (r >0.75) were determined by RT-qPCR. Additionally, glycosaminoglycans and collagen deposition were visualized with Alcian blue staining and immunohistochemistry (COL1 and COL2). Results Overexpression of TNFRSF11B resulted in strong upregulation of MMP13, COL2A1 and COL1A1. Likewise, mineralization and osteoblast characteristic markers RUNX2, ASPN and OGN showed a consistent increase. Among 30 genes highly correlated to TNFRSF11B, expression of only eight changed significantly, with BMP6 showing the highest increase (9-fold) while expression of RANK and RANKL remained unchanged indicating previously unknown downstream pathways of TNFRSF11B in cartilage. Conclusion Results of our 3D in vitro chondrogenesis model indicate that upregulation of TNFRSF11B in lesioned OA cartilage may act as a direct driving factor for chondrocyte to osteoblast transition observed in OA pathophysiology. This transition does not appear to act via the OPG/RANK/RANKL triad common in bone remodeling. Show less
Ben-Arzi, H.; A. das; Kelly, C.; Geest, R.J. van der; Plein, S.; Dall'Armellina, E. 2021
Background Four-dimensional (4D) flow cardiac magnetic resonance (cardiac MR) imaging provides quantification of intracavity left ventricular (LV) flow kinetic energy (KE) parameters in three... Show moreBackground Four-dimensional (4D) flow cardiac magnetic resonance (cardiac MR) imaging provides quantification of intracavity left ventricular (LV) flow kinetic energy (KE) parameters in three dimensions. ST-elevation myocardial infarction (STEMI) patients have been shown to have altered intracardiac blood flow compared to controls; however, how 4D flow parameters change over time has not been explored previously. Purpose Measure longitudinal changes in intraventricular flow post-STEMI and ascertain its predictive relevance of long-term cardiac remodeling. Study Type Prospective. Population Thirty-five STEMI patients (M:F = 26:9, aged 56 +/- 9 years). Field Strength/Sequence A 3 T/3D EPI-based, fast field echo (FFE) free-breathing 4D-flow sequence with retrospective cardiac gating. Assessment Serial imaging at 3-7 days (V1), 3-months (V2), and 12-months (V3) post-STEMI, including the following protocol: functional imaging for measuring volumes and 4D-flow for calculating parameters including systolic and peakE-wave LVKE, normalized to end-diastolic volume (iEDV) and stroke volume (iSV). Data were analyzed by H.B. (3 years experience). Patients were categorized into two groups: preserved ejection fraction (pEF, if EF > 50%) and reduced EF (rEF, if EF < 50%). Statistical Tests Independent sample t-tests were used to detect the statistical significance between any two cohorts. P < 0.05 was considered statistically significant. Results Across the cohort, systolic KEi(sv) was highest at V1 (28.0 +/- 4.4 mu J/mL). Patients with rEF retained significantly higher systolic KEi(sv) than patients with pEF at V2 (18.2 +/- 3.4 mu J/mL vs. 6.9 +/- 0.6 mu J/mL, P < 0.001) and V3 (21.6 +/- 5.1 mu J/mL vs. 7.4 +/- 0.9 mu J/mL, P < 0.001). Patients with pEF had significantly higher peakE-wave KEi(EDV) than rEF patients throughout the study (V1: 25.4 +/- 11.6 mu J/mL vs. 18.1 +/- 9.9 mu J/mL, P < 0.03, V2: 24.0 +/- 10.2 mu J/mL vs. 17.2 +/- 12.2 mu J/mL, P < 0.05, V3: 27.7 +/- 14.8 mu J/mL vs. 15.8 +/- 7.6 mu J/mL, P < 0.04). Data Conclusion Systolic KE increased acutely following MI; in patients with pEF, this decreased over 12 months, while patients with rEF, this remained raised. Compared to patients with pEF, persistently lower peakE-wave KE in rEF patients is suggestive of early and fixed impairment in diastolic function. Evidence Level 1 Technical Efficacy Stage 3 Show less
