Datasheet1_Prevention by the CXCR2 antagonist SCH527123 of the calcification of porcine heart valve cusps implanted subcutaneously in rats.docx

IntroductionCalcification is a main cause of bioprosthetic heart valves failure. It may be promoted by the inflammation developed in the glutaraldehyde (GA)-fixed cusps of the bioprosthesis. We tested the hypothesis that antagonizing the C-X-C chemokines receptor 2 (CXCR2) may prevent the calcification of GA-fixed porcine aortic valves.Materiel and methodsFour-week-old Sprague Dawley males were transplanted with 2 aortic valve cusps isolated from independent pigs and implanted into the dorsal wall. Four groups of 6 rats were compared: rats transplanted with GA-free or GA-fixed cusps and rats transplanted with GA-fixed cusps and treated with 1 mg/kg/day SCH5217123 (a CXCR2 antagonist) intraperitoneally (IP) or subcutaneously (SC) around the xenograft, for 14 days. Then, rats underwent blood count before xenografts have been explanted for histology and biochemistry analyses.ResultsA strong calcification of the xenografts was induced by GA pre-incubation. However, we observed a significant decrease in this effect in rats treated with SCH527123 IP or SC. Implantation of GA-fixed cusps was associated with a significant increase in the white blood cell count, an effect that was significantly prevented by SCH527123. In addition, the expression of the CD3, CD68 and CXCR2 markers was reduced in the GA-fixed cusps explanted from rats treated with SCH527123 as compared to those explanted from non-treated rats.ConclusionThe calcification of GA-fixed porcine aortic valve cusps implanted subcutaneously in rats was significantly prevented by antagonizing CXCR2 with SCH527123. This effect may partly result from an inhibition of the GA-induced infiltration of T-cells and macrophages into the xenograft.</p

Left ventricular contractile function of heart following electroporation mediated gene transfer using 8, 16 and 32 pulses protocols (n = 5 each group).

<p>Indices of contractility and relaxation determined respectively by the maximum (dP/dt max) (a) and the minimum (dP/dt min) (b) and developed LVP (dLVP) (c) of the first derivative of ventricular pressure with respect to time. These parameters remained unchanged before (time 0) and after electroporation except for the 32 pulses group where the values are transiently altered. A complete normalization of the values occurred within 10–20 minutes. No abnormality was detected in the group in which invivo electroporation was not performed.</p

The two early cardiac enzymes for injury TnT and CK-MB were measured; both enzymes remained within normal range after <i>in vivo</i> electroporation mediated gene transfer with 8 and 16 pulses, but the values were altered with the 32 pulses protocol.

<p>(a) TnT levels (b) CK-MB levels. The levels were below detection levels in the group in which invivo electroporation was not performed.</p

<i>In vivo</i> electroporation mediated gene transfer to the beating heart.

<p>(a) BLI images at day 1, 3 and 5 after <i>in vivo</i> electroporation mediated gene transfer to the normal heart. Graphical representation of the quantification of the luciferase activity, with electroporated compared to non-electroporated hearts; (b) BLI measurements, the photon counts per heart field are expressed in relative light units (RLU) 16479±4338 RLU for electroporated heart vs. 3539±1555 RLU for non electroporated heart (p = 0.018) at day 1. (c) Luciferase assay, RLU per milligram (mg) of protein was measured using a luminometer in the same sample 6594±1806 RLU/mg for electroporated heart vs 1684±760 RLU/mg of total protein for non electroporated heart (p = 0.011) at day 1. (d) Control heart, plasmid was injected but not elecroporated. (e) Expression of EGFP was found on the cell surface.</p

For histological analysis the hearts were evaluated at 24 hrs post gene transfer.

<p>The following criteria were considered: vascular congestion, infiltration and polymorphonuclear infiltrates. No haemorrhage or infiltration was noticed with any pulsing protocol. (a) 8 pulses (b) 16 pulses (c) 32 pulses (Magnification = ×200).</p

Graphical representation of transgene expression measured over time.

<p>After electroporation mediated gene transfer the animals were sacrificed at different time points.</p

The <i>in vivo</i> electroporation mediated gene transfer to the heart is a two step procedure.

<p>(a) First, the coronary sinus is occluded with a 6-0 prolene tourniquet and the plasmid is injected into the coronary vein. (b) The heart is then positioned between the plate electrodes and the electric field is applied.</p

MOESM2 of Biological scoring system for early prediction of acute bowel ischemia after cardiac surgery: the PALM score

Additional file 2: Table S2. Postoperative course

MOESM1 of Biological scoring system for early prediction of acute bowel ischemia after cardiac surgery: the PALM score

Additional file 1: Table S1. Type of cardiac surgery

MOESM3 of Biological scoring system for early prediction of acute bowel ischemia after cardiac surgery: the PALM score

Additional file 3: Table S3. Features for the distinction between mesenteric ischemia and ischemic colitis