Persistent atrial fibrillation without the evidence of low-voltage areas: a prospective randomized trial.

BACKGROUND: An effective therapy of persistent atrial fibrillation beyond pulmonary vein isolation remains unsatisfactory. Targeting endocardial low-voltage areas represents an approach of substrate modification. This prospective, randomized study investigated the efficacy of ablation of low-voltage areas versus PVI and additional linear ablations in patients with persistent atrial fibrillation in terms of single-procedure arrhythmia-free outcome and safety. METHODS AND RESULTS: A total number of 100 patients undergoing de-novo catheter ablation for persistent AF were randomized in a 1:1 ratio into two different treatment arms: group A: pulmonary vein isolation (PVI) and, if low-voltage areas were present, a substrate modification. Group B: PVI and, if atrial fibrillation persisted, additional ablations, such as linear ablation and/or ablation of non-PV triggers. A total of 50 patients were randomized into each group without significant differences in baseline characteristics. During a mean follow-up of 17.64 ± 4.5 months after a single procedure, 34 (68%) patients of group A were free of arrhythmia recurrence versus 28 (56%) patients in group B (p = ns). In group A, 30 (60%) patients did not show endocardial fibrosis and received solely PVI. Both procedures were performed with a low number of complications; no pericardial effusion or stroke were seen in either group. CONCLUSIONS: A significant proportion of patients with persistent atrial fibrillation do not show low-voltage areas. A total of 70% of the patients receiving solely PVI did not show any recurrence of atrial fibrillation, and therefore, extensive additional ablation should be avoided in de-novo patients.

Pulmonary vein isolation using cryoballoon ablation versus RF ablation using ablation index following the CLOSE protocol: A prospective randomized trial.

BACKGROUND: The single procedure success rates of durable pulmonary vein isolation (PVI) for paroxysmal atrial fibrillation (AF) vary between 80% and 90%. This prospective, randomized study investigated the efficacy of cryoballoon PVI (CBA) versus PVI with radio-frequency (RF)-energy following the CLOSE protocol (ablation index [AI], interlesion distance ≤6 mm, surround flow catheter) in terms of single-procedure arrhythmia-free outcome and safety. METHODS AND RESULTS: A total number of 150 patients undergoing de novo catheter ablation for paroxysmal AF were randomized to two different treatment arms. In group A patients, PVI was performed with the 23 or 28 mm cryoballoon (Artic Front™ Balloon in conjunction with an Achieve Mapping Catheter, Medtronic Inc.). The ablation procedure in group B was performed with RF-energy, using AI and following the CLOSE protocol. PVI using AI incorporates stability, contact force (CF), time, and power. The CLOSE protocol combines AI and ≤6 mm interlesion distance using a surround flow catheter (Biosense Webster Thermocool STSF). A total of 75 patients were randomized into each group without significant differences in baseline characteristics. During a mean follow-up of 12 ± 4.5 months after a single procedure, 64 (85.33%) patients of group A were free of arrhythmia recurrence versus 65 (86.67%) patients in group B (p = ns). A total of 14 patients (group A: 7 [9.33%]; group B: 7 [9.33%]; p = ns) underwent a redo-procedure. No significant difference between both groups was observed in terms of PV recovery (group A: 4 [5.33%] vs. group B: 3 [4%]; p = ns). In two patients of group A and four patients of group B, the PVs were durably isolated, whereas the patients had AF recurrence caused by extra-PV AF sources. Two patients of each group had continued paroxysmal AF but did not undergo redo-procedure. Patients of group A showed significantly more AF recurrence during the blanking period of 3 months (group A: 14 [18.67%] vs. group B: 6 [8%]; p < .05). With regard to the procedural data, the procedure time was significantly shorter in group A (70.53 ± 16.13 vs. 115.35 ± 15.38; p < .01); the flouroscopy time and dose area product showed no significant differences (Table 2). Both procedures were performed with a low number of complications; no pericardial effusion was seen in either group; in group A two patients had a significant hematoma of the groin with the need for surgical repair. CONCLUSIONS: Cryoballoon PVI and PVI using ablation index following the CLOSE protocol are equally efficient in achieving durable PV isolation. In this study, cryoballoon ablation led to significantly more AF recurrence during the blanking period.

Improved durable pulmonary vein isolation with shorter procedure times and lower energy levels using RF ablation with ablation index and a stringent lesion contiguity.

BACKGROUND: The single procedure success rates of durable pulmonary vein isolation (PVI) for paroxysmal atrial fibrillation (PAF) varies between 80 and 90%. Ablation index, incorporating contact force, stability, time and power is a more profound parameter of significant lesion size and has been established. Equally important is a stringent contiguity of the lesion set. METHODS AND RESULTS: A total number of 100 consecutive patients undergoing de-novo catheter ablation for paroxysmal atrial fibrillation (PAF) were analyzed between 2016 and 2019. In the first 50 patients (group A) PVI was performed using a surround flow, contact force catheter (Biosense Webster Thermocool STSF, Biosense Webster, USA) with a drag-and-ablate technique to encircle the PVs. In the following 50 patients (group B), PVI was performed using ablation index and a stringent lesion contiguity with an interlesion distance (ILD) of <5 mm. The baseline characteristics showed no significant differences between both groups. During a mean follow-up of 18 ± 3 months after a single procedure, 36 (72%) patients of group A were free of arrhythmia recurrence versus 43 (86%) patients in group B (p = 0.047). A total of 14 patients (group A: 10 (20%), group B: 4 (8%); underwent a redo-procedure. 7 patients of group A (14%) and 2 patients of group B (4%) showed recovered veins. In 3 patients of group A and 2 patients of group B the PVs were durably isolated. In these patients persistent AF recurrence was caused by extra-PV AF sources. Four patients of group A and three patients of group B had continued paroxysmal or persistent AF but did not undergo redo-procedure. With regard to the procedural data, the procedure time, the total energy and the fluoroscopy time were significantly lower in group B (AI and ILD <5 mm) (128.86 ± 18.19 versus 115.35 ± 15.38; p < 0.05; 1619.16 ± 988.56 versus 1186.26 ± 756.34; p < 0.05; 11.49 ± 3.20 versus 9.66 ± 3.86; p = 0.04). Both procedures were performed with a low number of complications, no pericardial effusion was seen in either group. CONCLUSIONS: PVI using ablation index in combination with a stringent lesion contiguity improves clinical outcome after first-time PVI with lower PVI recovery, shorter procedure times, lower total energy and shorter fluoroscopy times and therefore, is more efficient.

Implantation of leadless pacing systems in patients early after tricuspid valve surgery: A feasible option.

BACKGROUND: After tricuspid valve (TV) surgery due to tricuspid regurgitation (TR), patients needing a permanent pacemaker often receive an epicardial lead implantation. This may result in delayed recovery from open-chest surgery and increased postoperative risk. Leadless pacemaker (LPM) implantation may represent a valuable option. METHODS AND RESULTS: A total of 14 consecutive patients underwent LPM implantation (Micra Transcatheter Pacing System, Medtronic, Minneapolis, MN) early after TV surgery. The pacing indication in those patients was atrial fibrillation with a slow atrio-ventricular (AV) conduction or atrial fibrillation and a concomitant AV block III. Three patients already had a pacemaker prior to surgery, which was explanted during TV repair. Three patients received a valve replacement with a bioprosthesis, while the remaining eight patients received a TV repair. All procedural data and device measurements during and after LPM implantation were recorded. Transthoracic echocardiography was performed prior and post LPM implantation, showing no changes in TV or bioprosthesis performance. The device measurements were within an adequate range: threshold: 0.83 ± 0.34 V @ 0.24 ± 0 ms, impedance: 480 ± 58.88 ohm, and R-wave: 10.10 ± 3.60 mV. LPM implantation was successful in all patients with a mean procedural time of 32 ± 11.8 minutes, fluoroscopy time of 3.71 ± 3.15 minutes, and dose-area product of 536.67 ± 811.26 cGy/m2 . CONCLUSIONS: Implantation of an LPM early after TV surgery is a feasible option. LPM implantation does not affect TV or bioprosthesis performance in transthoracic echocardiography.

Possible influence of free fatty acid receptors on pH regulation in the ruminal epithelium of sheep.

High amounts of short-chain fatty acids (SCFAs) occur in the ovine rumen and constitute the animal's main energy source. However, they lead to an acidification of the ruminal epithelium. Therefore, effective intracellular pH (pHi ) regulation by transport proteins like monocarboxylate transporter 1 (MCT1) and Na+ /H+ exchangers (NHEs) is pivotal to ruminants to avoid epithelial damage. SCFAs might function not only as nutrients but also as signalling molecules by activating free fatty acid receptors (FFARs) in the ruminal epithelium and thus influence pHi regulation. FFARs work as nutrient sensors, transducing their information by modulating cyclic adenosine monophosphate (cAMP) levels. We hypothesized that (FFAR-modulated) decreases in cAMP levels stimulate the activity of MCT1 and NHEs in the ruminal epithelium of sheep. We detected two FFARs (GPR109A and FFAR2) immunohistochemically in the ovine ruminal epithelium. Administration of 10 mM butyrate to Ussing chamber-mounted epithelia provoked a significant reduction in intraepithelial cAMP levels. However, application of the GPR109A agonist niacin did not affect cAMP levels. MCT1 activity was analysed by measuring transepithelial 14 C-acetate fluxes, which were not inhibited by forskolin-induced increased cAMP levels. The recovery of pHi after acidification was assessed as an indicator of NHE activity in primary cultured ruminal epithelial cells. Recovery was significantly reduced when cells with increased cAMP levels were subjected to the NHE inhibitor 5-(N-ethyl-N-isopropyl)-amiloride (10 µM). Nonetheless, with augmented cAMP levels alone, NHE activity tended to decline. We hypothesize that modulation of cAMP levels by butyrate is accomplished by FFAR2 activation, regulating NHE activity for pHi homoeostasis at least in part.

Evaluation of the hepatocyte-derived cell line BFH12 as an in vitro model for bovine biotransformation.

The knowledge of drug metabolising enzymes (DMEs) in cattle is rather limited. The capability of the bovine foetal hepatocyte-derived cell line BFH12 to serve as model for biotransformation was evaluated. Gene expression analysis of DMEs was performed by reverse transcription PCR (RT-PCR). The presence of efflux transporters was visualised by immunocytochemistry, and functional induction of cytochrome P450 (CYP) 1A was assessed by the ethoxyresorufin-O-deethylase (EROD) assay. The production of bile acids was measured by liquid chromatography-tandem mass spectrometry (LC-MS/MS). RT-PCR revealed the expression of cytochromes 1A1, 1A2, 3A4 and phase II enzymes UGT1A1, UGT1A6 and GSTM1. Immunofluorescence demonstrated efflux transporters ABCG2 and ABCC1. The EROD assay revealed a dose-dependent CYP1A induction after treatment with benzo[a]pyrene (BP). LC-MS/MS analysis of cell culture supernatants showed the production of bile acids including taurocholic acid, tauro-chenodeoxycholic acid, taurodeoxycholic acid and taurolithocholic acid. The results strongly suggest the applicability of the cell line BFH12 for subsequent experiments in the emerging field of bovine biotransformation.

Establishment and Characterization of an SV40 Large T Antigen-Transduced Porcine Colonic Epithelial Cell Line.

Continuous cell lines have become indispensable tools that have enabled investigations into cellular mechanisms by increasing experimental reproducibility and sample availability, and decreasing the use of experimental animals. To facilitate studies of epithelial barrier function of the porcine colon, we aimed to establish an epithelial cell line with an extended replicative capacity. Cells were isolated from the proximal colon of a 3-week-old piglet and transduced using a recombinant retroviral vector construct containing the simian virus 40 large T antigen (SV40 TAg). We established a clonal epithelial cell line, referred to as PoCo83-3, that stably expressed the SV40 TAg, verified at mRNA and protein levels. PoCo83-3 showed epithelial cell-specific features, such as cobblestone-like morphology, dome structure formation, the presence of apical microvilli, and the expression of keratin 18, E-cadherin and the tight junction-associated proteins zonula occludens-1, occludin, and claudin-1. To validate PoCo83-3 as an in vitro model in epithelial barrier research, proinflammatory cytokine-inducible alterations in barrier integrity were demonstrated by incubating the cells with TNF-α and IFN-γ for 48 h. These cytokine treatments promoted a decreased transepithelial electrical resistance. In summary, PoCo83-3 exhibited an extended life span and a differentiated phenotype while maintaining epithelial characteristics. Based on these results, we present this cell line as a valuable in vitro model for investigations of epithelial barrier function in the porcine colon.

Establishment and characterisation of a novel bovine SV40 large T-antigen-transduced foetal hepatocyte-derived cell line.

Due to lack of in vitro models for bovine hepatocytes apart from primary cells, there is demand for a bovine hepatocyte-derived cell line. Transduction of bovine foetal hepatocytes with SV40 large T-antigen was performed using the vector pRetro-E2 SV40. Phase contrast microscopy was carried out to evaluate morphology. Immunofluorescence staining was conducted to study expression of keratins, tight junction proteins zona occludens-1 and claudin-1, glucose transporter-2 and P-glycoprotein as well as phosphoenolpyruvate carboxykinase. Urea and triglyceride production was quantified photometrically. Histochemical staining of glycogen by Periodic acid-Schiff stain and of lipids with Oil red O was performed after 24 h incubation with 20 mM glucose and 85 µM palmitic acid, respectively. Gene expression analysis of hepatocyte-typical genes was conducted by reverse transcription PCR. We obtained a SV40LTAg-transduced extended passage cell line, referred to as BFH12. Polygonal growth, keratins, tight junction proteins zona occludens-1 and claudin-1 and glucose transporter-2 as well as P-glycoprotein and phosphoenolpyruvate carboxykinase were attested positively. Urea production calculated as cell-specific rate was 14.2 ± 2.0 fmol/h (early passage) and 17.6 ± 3.7 fmol/h (late passage). Cell-specific triglyceride production was 1.6 ± 0.5 fmol/h (early passage) and 2.1 ± 0.3 fmol/h (late passage). Additionally, cells were positive for glycogen and lipid storage and showed a gene expression pattern resembling foetal hepatocytes. With the properties described here, the novel cell line BFH12 is a hepatocyte-derived cell line which can be used as an in vitro whole cell model.