TBX3 transfection and nodal signal pathway inhibition promote differentiation of adipose mesenchymal stem cell to cardiac pacemaker-like cells.

BACKGROUND: Mesenchymal stem cells (MSCs) are known as one of the best candidate cells to produce cardiac pacemaker-like cells (CPLCs). Upregulation of TBX3 transcription factor and inhibition of the nodal signal pathway have a significant role in the formation of cardiac pacemaker cells such as sinoatrial and atrioventricular nodes, which initiate the heartbeat and control the rhythm of heart contractions. This study aimed to confirm the effects of transfection of TBX3 transcription factor and inhibition of the nodal signal pathway on differentiating adipose-derived MSCs (AD-MSCs) to CPLCs. AD-MSCs were characterized using flow cytometry and three-lineage differentiation staining. METHODS: The transfection of TBX3 plasmid was carried out using lipofectamine, and inhibition of the nodal signal pathway was done using the small-molecule SB431542. The morphology of the cells was observed using a light microscope. Pacemaker-specific markers, including TBX3, Cx30, HCN4, HCN1, HCN3, and KCNN4, were evaluated using the qRT-PCR method. For protein level, TBX3 and Cx30 were evaluated using ELISA and immunofluorescence staining. The electrophysiology of cells was evaluated using a patch clamp. RESULTS: The TBX3 expression in the TBX3, SM, and TBX + SM groups significantly higher (p < 0.05) compared to the control group and cardiomyocytes. The expression of Cx40 and Cx43 genes were lower in TBX3, SM, TBX + SM groups. In contrast, Cx30 gene showed higher expression in TBX3 group. The expression HCN1, HCN3, and HCN4 genes are higher in TBX3 group. CONCLUSION: The transfection of TBX3 and inhibition of the nodal signal pathway by small-molecule SB431542 enhanced differentiation of AD-MSCs to CPLCs.

Progenitor Hematopoietic Cells Implantation Improves Functional Capacity of End Stage Coronary Artery Disease Patients with Advanced Heart Failure.

Background. Proangiogenic Hematopoietic Cells (PHC) which comprise diverse mixture of cell types are able to secrete proangiogenic factors and interesting candidate for cell therapy. The aim of this study was to seek for benefit in implantation of PHC on functional improvement in end stage coronary artery disease patients with advanced heart failure. Methods. Patients with symptomatic heart failure despite guideline directed medical therapy and LVEF less than 35% were included. Peripheral blood mononuclear cells were isolated, cultivated for 5 days, and then harvested. Flow cytometry and cell surface markers were used to characterize PHC. The PHC were delivered retrogradely via sinus coronarius. Echocardiography, myocardial perfusion, and clinical and functional data were analyzed up to 1-year observation. Results. Of 30 patients (56.4 ± 7.40 yo) preimplant NT proBNP level is 5124.5 ± 4682.50 pmol/L. Harvested cells characterized with CD133, CD34, CD45, and KDR showed 0.87 ± 0.41, 0.63 ± 0.66, 99.00 ± 2.60, and 3.22 ± 3.79%, respectively. LVEF was improved (22 ± 5.68 versus 26.8 ± 7.93, p < 0.001) during short and long term observation. Myocardial perfusion significantly improved 6 months after treatment. NYHA Class and six-minute walk test are improved during short term and long term follow-up. Conclusion. Expanded peripheral blood PHC implantation using retrograde delivery approach improved LV systolic function, myocardial perfusion, and functional capacity.

Adult congenital cardiac surgery in Indonesia.

BACKGROUND: Successful paediatric cardiac surgery and cardiology treatment has resulted in an increase in the use of surgery as a method of treatment of congenital cardiac disease in adult population. However, late detection and lower socio-economic condition in developing countries might change patients' characteristics by the time they come for treatment. This study aimed to elaborate the long-term surgical results of adult congenital cardiac disease in Indonesia as a developing country. METHODS AND RESULTS: We reviewed retrospectively all adult congenital cardiac disease patients with a mean age of 28 years plus or minus 9.5 years, who underwent surgery at National Cardiovascular Center. The types of procedures used were corrective in 338 patients (89.2%), palliative in 10 patients (2.6%), and re-operations in 31 patients (8.2%). The overall hospital mortality rate was 2.6% but as high as 20% with palliative surgery. Post-operative New York Heart Association class III-IV is the only independent predictor of death at 60 months (hazard ratio 61.48, 95% confidence interval 9.41-401.69, p<0.001). The survival rates were 96.3% and 95% for overall and non-atrial septal defect in patients at 60 months, which was highest in corrective procedures (97.6%). The percentage of patients free of re-operation at 5 years' follow-up was 85.4% and 42.7% at 10 years. CONCLUSION: In developing countries, surgical treatment of adult congenital cardiac disease is effective and safe, with an overall survival rate of 96.3% at 60 months. Due to high mortality rate, palliative surgery of a non-atrial septal defect patient is recommended to be discontinued. The independent predictor of mortality was post-operative New York Heart Association functional class III-IV.

Mechanism of adenosine-induced termination of focal atrial tachycardia.

INTRODUCTION: Adenosine can terminate most focal atrial tachycardias (ATs). However, information about the termination mechanism is limited. This study investigated the effects and mechanism of adenosine on terminating focal AT using a three-dimensional noncontact mapping system. METHODS AND RESULTS: The study consisted of 7 patients (4 men and 3 women; age 44 +/- 29 years) with focal AT. Cycle length variation and atrial activation pattern at baseline and just before AT termination by adenosine (3-12 mg) were analyzed. Noncontact mapping demonstrated focal AT propagated from the origin (O) with preferential conduction and spread away from the breakout sites to the whole atrium. Compared to baseline AT, termination episodes revealed higher mean beat-to-beat variation of AT cycle length (11.7 +/- 11.7 msec vs 4.7 +/- 4.5 msec, P < 0.001) and standard deviation of normalized AT cycle length (0.033 +/- 0.014 vs 0.011 +/- 0.005, P < 0.001). In termination episodes, adenosine significantly decreased the peak negative voltage of AT-O (-27.2 +/- 15.3%, P < 0.01), preferential conduction (proximal: -32.1 +/- 18.7, mid: -28.4 +/- 22.8, distal portion: -29.6 +/- 21.4%, P < 0.01), and breakout (-31.4 +/- 12.5%, P < 0.01). However, adenosine did not affect voltage in nontermination episodes. Adenosine shifted the locations of AT-O in 5 of 10 AT episodes with termination. Mean number of shifting AT-O was 2.4 +/- 1.5 (range 1-4), with maximum shifting distance of 15.0 +/- 3.1 (range 10-19) mm. Focal activation at AT-O simply disappeared in all termination episodes and therefore was not due to conduction block within preferential conduction or breakout site. Catheter ablation lesions covered 50% of total shifting origins, without late recurrence. CONCLUSION: Adenosine-induced AT termination was associated with significantly decreased electrogram voltage, shifting AT-O locations, and disappearance of focal activation.

Focal atrial tachycardia: new insight from noncontact mapping and catheter ablation.

BACKGROUND: This study investigated the electrophysiologic characteristics, atrial activation pattern, and effects of radiofrequency (RF) catheter ablation guided by noncontact mapping system in patients with focal atrial tachycardia (AT). METHODS AND RESULTS: In 13 patients with 14 focal ATs, noncontact mapping system was used to map and guide ablation of AT. AT origins were in the crista terminalis (n=8), right atrial (RA) free wall (n=3), Koch triangle (n=1), anterior portion of RA-inferior vena cava junction (n=1), and superior portion of tricuspid annulus (n=1); breakout sites were in the crista terminalis (n=5), RA free wall (n=5), middle cavotricuspid isthmus (n=2), and RA-superior vena cava junction (n=2). ATs arose from the focal origins (11 ATs inside or at the border of low-voltage zone), with preferential conduction, breakout, and spread to the whole atrium. After applications of RF energy on the earliest activation site or the proximal portion of preferential conduction from AT origin, 13 ATs were eliminated without complication. During the follow-up period (8+/-5 months), 11 (91.7%) of the 12 patients with successful ablation were free of focal ATs. CONCLUSIONS: Focal AT originates from a small area and spreads out to the whole atrium through a preferential conduction. Application of RF energy guided by noncontact mapping system was effective and safe in eliminating focal AT.