142 Intravital imaging of leukocyte, platelet and stem cell trafficking in vivo in the cardiac microcirculation following myocardial ischaemia-reperfusion injury

Abstract

infarct size was observed when WT-MI hearts were compared to their PMCA1Ht counterparts 1 week post-surgery. Interestingly, whilst both WT and PMCA1Ht MI-treated mice showed a significant deterioration in cardiac function 1 week post-surgery there were differences seen in cardiac structure between the two groups. For example, echocardiography revealed that the WT-MI hearts were significantly more hypertrophic when compared to their PMCA1Ht counterparts shown by a significant increase in the LV diameters alongside a significant difference in the normalised heart weight to tibia length ratios. ECG revealed significantly longer QT intervals and considerably more extra-systolic events among the WT-MI mice. Whilst histological analysis of cardiomyocyte size showed a significantly exacerbated hypertrophic response in the remote regions of WT-MI mice compared to PMCA1 Ht -MI mice. Conclusion Heterozygous deletion of PMCA1 might serve a protective role in the heart following both moderate and severe MI. The protective mechanism most likely develops in the early post-operative phase, as an MI of similar extent is associated with higher mortality rates in WT mice. Future work will aim to elucidate the mechanism producing this phenotype mainly focusing on the potential role that PMCA1 might serve in the process. Introduction Lipids have key roles in CVD but heritability studies focus on few species bar lipoproteins. Lipids play key roles in cell signalling, immunity, inflammation, vasodilation and cell death. Although not encoded, their activities are tightly linked to DNA-encoded entities (e.g. enzymes and other proteins) and those with a strong genetic influence (high heritability) may identify novel pathways in CVD. Purpose: Analysing eicosanoid, endocannabinoid and sphingolipid profiles in 250 British Caucasian families with GWAS data will identify particularly heritable lipid biomarkers for the discovery of causative genetic variation of CVD. Methods An array of 79 eicosanoids and related species, 33 endocannabinoids and congeners, 63 ceramides and related species, from 204 plasma samples (31 families of 1+ individuals with hypertension) were extracted and analysed by liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). Quality control was accessed and estimates of heritability of each lipid species was estimated using QTDT software. The concentration of each lipid species were assessed for normal distribution, outliers, and adjusted for age and sex. The resulting lipid concentrations will be analysed using FaST-LMM software for associations with 557,124 SNPs. Results 19 species of eicosanoids were identified (mean concentrations 19 pg/ml -7600 pg/ml); the species at highest abundance in plasma (HODEs) are derivatives of the omega-6 fatty acid linoleic acid. 8 species passed quality control assessments and 3 species were estimated to be significantly heritable (9-OxoODE 24%, 12-HETE 44%, 12(13)-EPOME 78%). 15 congeners of endocannabinoids were identified (mean concentrations 20 pg/ml -4000 pg/ml); the...

Tify: A quality-based frame selection tool for improving the output of unstable biomedical imaging

“…Tify was tested using images obtained from existing intravital microscopy studies that had taken place in our laboratory[ 12 ]. C57BL/6 mice (obtained from Envigo at 8–12 weeks age) were anaesthetised using ketamine/medetomidine (100mg/kg, 10mg/kg, respectively).…”