Phosphorylcholine antibodies restrict infarct size and left ventricular remodelling by attenuating the unreperfused post-ischaemic inflammatory response

Phosphorylcholine is a pro-inflammatory epitope exposed on apoptotic cells, and phosphorylcholine monoclonal immunoglobulin (Ig)G antibodies (PC-mAb) have anti-inflammatory properties. In this study, we hypothesize that PC-mAb treatment reduces adverse cardiac remodelling and infarct size (IS) following unreperfused transmural myocardial infarction (MI). Unreperfused MI was induced by permanent ligation of the left anterior descending (LAD) coronary artery in hypercholesterolaemic APOE*3-Leiden mice. Three weeks following MI, cardiac magnetic resonance (CMR) imaging showed a reduced LV end-diastolic volume (EDV) by 21% and IS by 31% upon PC-mAb treatment as compared to the vehicle control group. In addition, the LV fibrous content was decreased by 27% and LV wall thickness was better preserved by 47% as determined by histological analysis. Two days following MI, CCL2 concentrations, assessed by use of ELISA, were decreased by 81% and circulating monocytes by 64% as assessed by use of FACS analysis. Additionally, local leucocyte infiltration determined by immunohistological analysis showed a 62% decrease after three weeks. In conclusion, the local and systemic inflammatory responses are limited by PC-mAb treatment resulting in restricted adverse cardiac remodelling and IS following unreperfused MI. This indicates that PC-mAb holds promise as a therapeutic agent following MI limiting adverse cardiac remodelling

Utilization of neuronavigation in the influence on the visual cortex with transcranial magnetic stimulation (TMS)

The topic of my thesis I chose because of my interest in the fields psychiatry and neurology, mainly new possibilities for treatment and diagnosis of diseases and to a great extent due to the possibility of being part of a research project doc. George Horacek Prague Psychiatric Cente

Hypercholesterolemia affects cardiac function, infarct size and inflammation in APOE*3-Leiden mice following myocardial ischemia-reperfusion injury.

BackgroundHypercholesterolemia is a major risk factor for ischemic heart disease including acute myocardial infarction. However, long-term effects of hypercholesterolemia in a rodent myocardial ischemia-reperfusion injury model are unknown. Therefore, the effects of diet-induced hypercholesterolemia on cardiac function and remodeling were investigated up to eight weeks after myocardial ischemia-reperfusion (MI-R) injury which was induced in either normocholesterolemic (NC-MI) or hypercholesterolemic (HC-MI) APOE*3-Leiden mice.MethodsLeft ventricular (LV) dimensions were serially assessed using parasternal long-axis echocardiography followed by LV pressure-volume measurements. Subsequently, infarct size and the inflammatory response were analyzed by histology and fluorescence-activated cell sorting (FACS) analysis.ResultsIntrinsic LV function eight weeks after MI-R was significantly impaired in HC-MI compared to NC-MI mice as assessed by end-systolic pressure, dP/dtMAX, and -dP/dtMIN. Paradoxically, infarct size was significantly decreased in HC-MI compared to NC-MI mice, accompanied by an increased wall thickness. Hypercholesterolemia caused a pre-ischemic peripheral monocytosis, in particular of Ly-6Chi monocytes whereas accumulation of macrophages in the ischemic-reperfused myocardium of HC-MI mice was decreased.ConclusionDiet-induced hypercholesterolemia caused impaired LV function eight weeks after MI-R injury despite a reduced post-ischemic infarct size. This was preceded by a pre-ischemic peripheral monocytosis, while there was a suppressed accumulation of inflammatory cells in the ischemic-reperfused myocardium after eight weeks. This experimental model using hypercholesterolemic APOE*3-Leiden mice exposed to MI-R seems suitable to study novel cardioprotective therapies in a more clinically relevant animal model

Characterization of primary cultured MSCs.

<p>A: <i>In vitro</i> characterization of MSCs consisting of the specific surface marker antigen panel measured by flow cytometry. B: The differentiation capacity of MSCs towards osteoblasts shown by alkaline phosphatase activity and towards adipocytes shown by lipid droplets staining via Oil Red O. C: The inhibitory capacity of proliferation of activated peripheral blood mononuclear cells measured by 3H-thymidine uptake in counts per minute (CCPM). D: Bright light and fluorescence microscopy and flow cytometric analysis of eGFP labeling of lentivirally transduced MSCs.</p

Flow cytometric analysis of inflammatory cells in the heart 15 days after myocardial infarction in uMSC and iMSC treated animals (uMSC and iMSC, resp.), PBS treated animals (PBS) and Sham-operated animals (Sham).

<p>Data are expressed as mean ± SD. There were no significant differences between the uMSC, iMSC, PBS and Sham group.</p

Stimulation of MSCs with the pro-inflammatory cytokine interferon gamma (IFN).

<p>A: From left to right: Immunostaining of iMSCs for HLA-DR in red (PE) and nuclei in blue (DAPI) and expression of HLA-DR as measured by flow cytometry. B: Immunostaining of iMSCs for the immunomodulatory enzyme IDO in green (ALEXA 488 nm) and nuclei in blue (DAPI).</p

Cardiac function as assessed by 7 T MRI at 2 and 14 days after myocardial infarction in uMSC and iMSC treated animals (uMSC and iMSC, resp.), PBS treated animals (PBS) and Sham-operated animals (Sham) (panel A-C).

<p>A: Left ventricular ejection fraction. B: End-diastolic volume. C: End-systolic volume. Data are expressed as mean ± SD. # = p<0.05 versus Sham. D: Pressure Volume loops of all treatment groups at day 15 after MI. The oblique lines represent the end-systolic (Ees) and end-diastolic (Eed) pressure–volume relations.</p

Physical parameters 15 days after myocardial infarction in uMSC and iMSC treated animals (uMSC and iMSC, resp.), PBS treated animals (PBS) and Sham-operated animals (Sham).

<p>A: Weight loss. B: Amount of pulmonary fluid. Data are expressed as mean ± SD. There were no significant differences between the uMSC, iMSC, PBS and Sham group.</p

Assessment of engraftment of injected MSCs in mouse hearts 15 days after injection into the infarcted myocardium.

<p>A: Immunofluorescent staining of engrafted eGFP-labeled MSCs in red (Qdot 655) and nuclei in blue (hoechst 33342). B: Histological quantification of uMSC and iMSC engraftment (uMSC and iMSC resp.). PBS treated animals (PBS) and Sham-operated animals (Sham) were used as controls. C: Quantitative PCR for human genomic DNA in mouse hearts in animals treated with uMSC, iMSC, PBS and sham-operated animals. Data are expressed as mean ± SD. There were no significant differences between the uMSC and iMSC group.</p