The UHRF1 protein is a key regulator of retrotransposable elements and innate immune response to viral RNA in human cells.

While epigenetic mechanisms such as DNA methylation and histone modification are known to be important for gene suppression, relatively little is still understood about the interplay between these systems. The UHRF1 protein can interact with both DNA methylation and repressive chromatin marks, but its primary function in humans has been unclear. To determine what that was, we first established stable UHRF1 knockdowns (KD) in normal, immortalized human fibroblasts using targeting shRNA, since CRISPR knockouts (KO) were lethal. Although these showed a loss of DNA methylation across the whole genome, transcriptional changes were dominated by the activation of genes involved in innate immune signalling, consistent with the presence of viral RNA from retrotransposable elements (REs). We confirmed using mechanistic approaches that 1) REs were demethylated and transcriptionally activated; 2) this was accompanied by activation of interferons and interferon-stimulated genes and 3) the pathway was conserved across other adult cell types. Restoring UHRF1 in either transient or stable KD systems could abrogate RE reactivation and the interferon response. Notably, UHRF1 itself could also re-impose RE suppression independent of DNA methylation, but not if the protein contained point mutations affecting histone 3 with trimethylated lysine 9 (H3K9me3) binding. Our results therefore show for the first time that UHRF1 can act as a key regulator of retrotransposon silencing independent of DNA methylation.

ArrayNinja: An Open Source Platform for Unified Planning and Analysis of Microarray Experiments.

Microarray-based proteomic platforms have emerged as valuable tools for studying various aspects of protein function, particularly in the field of chromatin biochemistry. Microarray technology itself is largely unrestricted in regard to printable material and platform design, and efficient multidimensional optimization of assay parameters requires fluidity in the design and analysis of custom print layouts. This motivates the need for streamlined software infrastructure that facilitates the combined planning and analysis of custom microarray experiments. To this end, we have developed ArrayNinja as a portable, open source, and interactive application that unifies the planning and visualization of microarray experiments and provides maximum flexibility to end users. Array experiments can be planned, stored to a private database, and merged with the imaged results for a level of data interaction and centralization that is not currently attainable with available microarray informatics tools.

Substrate Specificity Profiling of Histone-Modifying Enzymes by Peptide Microarray.

The dynamic addition and removal of covalent posttranslational modifications (PTMs) on histone proteins serves as a major mechanism regulating chromatin-templated biological processes in eukaryotic genomes. Histone PTMs and their combinations function by directly altering the physical structure of chromatin and as rheostats for effector protein interactions. In this chapter, we detail microarray-based methods for analyzing the substrate specificity of lysine methyltransferase and demethylase enzymes on immobilized synthetic histone peptides. Consistent with the "histone code" hypothesis, we reveal a strong influence of adjacent and, surprisingly, distant histone PTMs on the ability of histone-modifying enzymes to methylate or demethylate their substrates. This platform will greatly facilitate future investigations into histone substrate specificity and mechanisms of PTM signaling that regulate the catalytic properties of histone-modifying enzymes.

Instantaneous lead entrapment: an unusual complication of nonthoracotomy implantation of an endocardial defibrillation lead.

Nonthoracotomy implantation of implantable cardioverter defibrillators is performed with transvenous leads that are similar to pacemaker leads and are subject to the same potential problems. We report an unusual complication of lead placement in which an electrode immediately became entrapped in the superior rim of the tricuspid valve, resisting all efforts at removal.

Safety, feasibility and long term follow-up of a non-thoracotomy defibrillation system in patients with inducible sustained ventricular tachycardia/fibrillation.

UNLABELLED: A non-thoracotomy lead system CPI-ENDOTAK, a transvenous lead used alone or combined with a subcutaneous patch (SQ-P), was evaluated as an alternative to epicardial patches/electrodes in patients at high risk for sudden cardiac death undergoing implantable cardioverter-defibrillator (ICD) surgery. Fifty nine patients, 62 +/- 11.4 years with CAD (83.0%) cardiomyopathy (11.9%) other (5.1%), mean ejection fraction 31.8 +/- 14%, with inducible sustained VT/VF underwent testing of either lead alone or lead/SQ-P. Four configurations of NTL were tested. Fifty one patients had NTL implanted (lead alone = 60.8% and lead/SQ-P = 39.2%). Eight patients required non-NTL approaches, due to high DFT (7) or anatomic anomaly (1). DFT's were 19.1J (lead alone) and 20.8J (lead/SQ-P). Acute complications: pulmonary embolism 1, lead dislodgement 3, sensing malfunction 1. [table: see text] CONCLUSION: A NTL system using either a single transvenous lead alone or combined with SQ-P can be implanted successfully in high risk patients with a low incidence of acute complications. Non-arrhythmic survival is lowest in patients receiving defibrillation shocks. Arrhythmic survival is high in all patients.

Prospective evaluation of a sequential pacing and high-energy bidirectional shock algorithm for transvenous cardioversion in patients with ventricular tachycardia.

Rapid ventricular pacing alone or in combination with low- or intermediate-energy shocks has limited efficacy in cardioverting rapid ventricular tachycardia (VT) when delivered through two transvenous catheter electrodes. This prospective study determined the efficacy and safety of an algorithm that used a sequence of rapid ventricular pacing (RVP) and intermediate-energy (5 and 15 J) and high-energy (25J) single, bidirectional shocks delivered by two transvenous catheter electrodes in conjunction with a cutaneous electrode in patients with sustained VT. The bidirectional shock was simultaneously delivered over two electrical vectors via a common right ventricular apical cathode and tow anodes consisting of the superior vena caval catheter electrode and cutaneous patch. The electrical therapy delivered was determined by the cycle length of VT. Slow VT (cycle length greater than 300 msec) was sequentially treated by RVP followed by incremental bidirectional shocks of 5, 15, and 25 J. Rapid VT (cycle length less than 300 msec) was treated with no incremental bidirectional shocks of 15 and 25 J. VT was reinduced to determine reproducibility of the algorithm for episodes that were successfully terminated. For patients in whom the primary algorithm failed, a second algorithm was used that excluded 5 and 15 J shocks and went directly to a 25 J shock. VT was reinduced twice and the secondary algorithm was evaluated. Thus, reproducibility of termination of VT with the primary and secondary algorithm was examined. Fifty episodes of slow VT and 40 episodes of rapid VT were induced in 22 patients (mean left ventricular ejection fraction 31 +/- 14%). Six patients had rapid VT, nine patients had slow VT, and seven patients had both.(ABSTRACT TRUNCATED AT 250 WORDS)

A prospective evaluation of single and dual current pathways for transvenous cardioversion in rapid ventricular tachycardia.

By using a prospective randomized study design, we compared the clinical efficacy and safety of single unidirectional and bidirectional transvenous cardioversion shocks for termination of rapid ventricular tachycardia (VT) having cycle lengths less than 300 ms. A Medtronic 6880 catheter was placed in the right ventricular apex and an R2 skin patch electrode was placed over the left scapula. Patients were randomized into two groups. Group A patients received unidirectional transvenous shocks using the two catheter electrodes (right ventricular apical cathode and superior vena caval anode) which resulted in a single current pathway. Group B patients received bidirectional transvenous shocks using a common cathode (right ventricular apex) and two separate anodes (superior vena caval and R2 patch) resulting in two current pathways. Identical shocks with total energies of 2.7, 5.0 and 10.0 J and waveform tilt of 27% were delivered to Groups A and B. In selected Group B patients, delivered shock currents through the right ventricular apex/superior vena caval and right ventricular apex/R2 patch electrode pairs were measured. We analyzed the initial episode of VT with a cycle length less than 300 ms in 33 patients with organic heart disease (mean age, 64 +/- 9 years; mean VT cycle length, 248 +/- 37 ms) who underwent programmed electrical stimulation. Transvenous cardioversion shocks terminated 31% of 16 VT episodes in Group A and 41% of 17 VT episodes in Group B (p greater than .2). The mean successful shock energy was 6.1 +/- 3.7 J in Group A and 3.0 +/- 0.9 J in Group B (p less than .05). Forty percent of all successfully cardioverted episodes in Group A and 86% of all successfully cardioverted VT episodes in Group B were terminated at an energy of 2.7 J (p = .09). Analysis of shock waveforms in Group B revealed 47 to 74% of the total current was transmitted through the right ventricular apex/superior vena caval electrodes and 26 to 53% through the right ventricular apex/R2 electrodes. We conclude that single bidirectional transvenous shocks are effective for rapid VT termination in selected patients. Dual current pathways decrease energies needed for successful transvenous cardioversion in this patient population.

A special outpatient clinic for following patients with implanted tachyarrhythmia devices.

Implantable anti-tachycardia devices have become an additional therapeutic option for those patients afflicted with life-threatening tachyarrhythmias. Follow-up of these complex devices are time-consuming and, if mismanaged, may be dangerous to the patient. For these reasons, a special anti-tachycardia device clinic was started at Newark Beth Israel Medical Center in July 1984. From the inception of the clinic to September 1985, 24 patients were followed. Seventy-five percent had antitachycardia devices (ATDs) implanted for treatment of ventricular tachyarrhythmias (VT/VF) with the remaining 25% for supraventricular tachycardias. All patients were seen every 3 months or more often if clinically required. Of 112 clinic examinations, 102 (91%) were scheduled appointments (group I) while the remaining 10 visits (group II) were unscheduled and preceded by symptomatic episodes. The problems detected in clinic (groups I and II) ranged from sudden failure of an AICD to apprehension. Appropriate nonoperative treatment was given during clinic evaluation for 60% of the problems detected in group II, while the remaining 40% required eventual surgical intervention. Compliance throughout the 15-month follow-up period was 100%. Major benefits of the clinic cited by patients and their families were continuity of care, the time allotted to meet the individual needs, and management of most problems on an out-patient basis.

Long-term efficacy and safety of beta-adrenergic receptor antagonists for supraventricular tachycardia.

The efficacy and safety of nadolol and atenolol, 2 new long-acting beta-adrenergic receptor antagonists, were evaluated in patients with recurrent supraventricular tachycardia (SVT). Intravenous and oral drug therapy was administered to patients with atrioventricular reentrant tachycardia and atrioventricular nodal reentrant tachycardia. Efficacy was judged on a short-term basis by programmed electrical stimulation and on a long-term basis by clinical parameters and serial ambulatory electrocardiographic recordings during long-term follow-up. In addition, the usefulness of programmed electrical stimulation to predict long-term efficacy was evaluated. Intravenous nadolol prevented induction of SVT in 6 of 8 (75%) patients with atrioventricular nodal reentrant tachycardia, and oral nadolol prevented induction of SVT in 5 of 6 (83%) responders to intravenous nadolol. No episodes of sustained SVT recurred in these 5 patients during follow-up. Intravenous nadolol also prevented induction of SVT in 2 of 17 (11%) patients with atrioventricular reentrant tachycardia. Both patients remained non-inducible during treatment with oral nadolol, and neither experienced recurrence of SVT during follow-up. Intravenous atenolol prevented induction of SVT in 5 of 6 (83%) patients with atrioventricular nodal reentrant tachycardia. Oral atenolol prevented induction of atrioventricular nodal reentrant tachycardia in 4 of 5 (80%) patients responding to intravenous atenolol, and none of these 4 patients experienced a clinical recurrence. Intravenous atenolol prevented induction of SVT in 1 of 4 (25%) patients with atrioventricular reentrant tachycardia. Oral atenolol prevented induction of SVT in this patient and the arrhythmia has not recurred during follow-up. During follow-up (1 to 37 months), drug tolerance and compliance have been excellent with a low incidence of adverse reactions (11%).(ABSTRACT TRUNCATED AT 250 WORDS)

Laser ablation of normal and diseased human ventricle.

The feasibility and safety of laser photoablation in patients with ventricular tachycardia (VT) and accessory pathways are currently being examined. We studied the qualitative and quantitative effects of argon laser radiation on normal and diseased human ventricle to determine the relationship between the size of tissue lesion and delivered energy. Twenty-nine human ventricle segments (normal ventricle = 10; diseased ventricle = 19) were excised from patients during mapping-guided subendocardial resection for VT (seven patients), mitral valve replacement (five patients), or immediately at autopsy (three patients). Lasing was performed with a 15 W argon laser coupled to a 300 micron optical fiber. Incremental laser discharges from 10 to 1000 J were delivered in air and saline with the optical fiber 5 mm from the endocardial surface. Gross and microscopic damage was quantified and correlated with laser discharges at low (10 to 100 J), intermediate (101 to 300 J), and high (greater than 300 J) energies. Histologic examination of laser-induced lesions in both normal and diseased human ventricle in either medium showed focal thermal injury with crater formation, vacuolization, and coagulation necrosis of endocardium and myocardium. In normal ventricle, mean lesion diameter and depth in air increased with increasing energies up to 300 J. Over 300 J, tissue perforation was frequently observed. In saline, the mean lesion depth was significantly reduced (p less than 0.02) at comparable energies. In diseased ventricle, mean lesion diameter and depth in air and saline also increased with increasing laser discharge energies up to 300 J. Higher energy laser discharges did not increase mean lesion dimensions or result in tissue perforation.(ABSTRACT TRUNCATED AT 250 WORDS)

Usefulness of an implantable antitachycardia pacemaker system for supraventricular or ventricular tachycardia.

The Cordis Omni-Orthocor model 234A, an implantable antitachycardia system, was evaluated in 13 patients. Two patients had recurrent sustained supraventricular tachycardia (SVT) and 11 had ventricular tachycardia (VT). The system was used for SVT or VT termination (group 1: SVT, 2 patients; VT, 4 patients) or for demand pacing and noninvasive electrophysiologic studies for tachycardia induction and serial electrophysiologic testing alone (group 2: VT, 7 patients). The overdriver was used successfully in 4 of 6 patients in group 1 for repeated tachycardia termination (SVT and VT) during a mean follow-up period of 18 months. One patient had 1 sustained VT episode unresponsive to pacing and 1 patient had no recurrence of tachycardia. Tachycardia termination zones varied when using the system in 2 patients receiving long-term amiodarone therapy. Eighteen noninvasive electrophysiologic studies for serial drug testing were performed, 4 in group 1 and 14 in group 2. Clinical tachycardia was induced and successfully terminated by use of the overdriver in 12 studies. It is concluded that implantable antitachycardia systems can be used successfully for noninvasive tachycardia induction and termination and for reliable serial electrophysiologic studies. Such systems provide improved patient safety and acceptability and are reasonable in cost.

Clinical electrophysiology, efficacy and safety of chronic oral cibenzoline therapy in refractory ventricular tachycardia.

The electrocardiographic (ECG) and electrophysiologic (EP) effects, clinical efficacy and safety of oral cibenzoline therapy were evaluated using a twice-daily dosing regimen in patients with refractory ventricular tachycardia (VT). Twenty patients underwent EP studies in the control (drug-free) state and after cibenzoline therapy using an incremental dose-titration protocol. Oral cibenzoline (2.4 to 5.8 mg/kg/day) was administered in doses of 130, 160 or 190 mg at 12-hour intervals. ECG and EP variables, 24-hour ambulatory ECG monitoring and programmed electrical stimulation studies were obtained in the control state and after 11 +/- 4 days of cibenzoline therapy. Cibenzoline therapy prolonged the mean PR interval (from 179 +/- 29 to 201 +/- 36 ms, p less than 0.001), the mean QRS duration (from 107 +/- 21 to 130 +/- 25 ms, p less than 0.001), and the mean QTc interval (from 422 +/- 25 to 460 +/- 42 ms, p less than 0.001). It increased the mean HV interval (from 50 +/- 17 to 65 +/- 20 ms, p less than 0.01) and mean right ventricular effective refractory period (from 245 +/- 24 to 266 +/- 27 ms, p less than 0.01). After cibenzoline therapy, 5 patients (25%) had suppression of inducible sustained VT during programmed electrical stimulation. High-degree atrioventricular block occurred in 2 patients. Chronic cibenzoline therapy (mean follow-up 24 +/- 3 months) remained effective in long-term suppression of VT in 4 patients. Two patients had to discontinue therapy because of gastrointestinal intolerance. Cibenzoline is effective in suppression of refractory VT in selected patients using a twice-daily dosing schedule.

Comparative efficacy of transvenous cardioversion and pacing in patients with sustained ventricular tachycardia: a prospective, randomized, crossover study.

We performed a prospective, randomized crossover study to evaluate the comparative efficacy of transvenous cardioversion and rapid ventricular pacing for termination of induced ventricular tachycardia in patients with spontaneous ventricular tachycardia and organic heart disease. Sixty-two episodes of ventricular tachycardia were induced in 15 patients, mean age 60 +/- 10 years, during electrophysiologic studies. All patients underwent a preselected electrical therapy protocol in a randomized crossover sequence. Transvenous cardioversion was performed by an incremental protocol of three sequential shocks (0.5, 1.1, and 2.7 J). Six asynchronous sequential bursts of rapid ventricular pacing (10 and 15 paced stimuli at 90%, 75%, and 65% of ventricular tachycardia cycle length) were used. Mean cycle length of ventricular tachycardia for the study population was 391 +/- 85 msec. The morphology of the tachycardia was left bundle branch block in 27, right bundle branch block in 32, and indeterminate in three. Characteristics of ventricular tachycardia terminated by the two techniques were comparable. Rate of success for termination of tachycardia with the two methods was also comparable (transvenous cardioversion 83%, rapid ventricular pacing 80%; p greater than .1) and these responses were concordant in 78%. The modes of termination of ventricular tachycardia were similar. The incidence of acceleration of ventricular tachycardia per episode with these preselected protocols was also comparable (transvenous cardioversion 11%, rapid ventricular pacing 6%; p greater than .2). Transient supraventricular tachyarrhythmias were more frequent after transvenous cardioversion (23%) than after rapid ventricular pacing (3%). Significant patient discomfort occurred only after transvenous cardioversion (incidence of 57%).(ABSTRACT TRUNCATED AT 250 WORDS)

Studies on left ventricular function during sustained ventricular tachycardia.

The acute effects of rapid ventricular pacing and sustained ventricular tachycardia on left ventricular function were examined in patients with recurrent sustained ventricular tachycardia. Programmed electrical stimulation and left ventricular hemodynamic measurements were performed in 20 patients (19 men and 1 woman), with an age range of 49 to 79 years (mean 63 +/- 9). Indexes of left ventricular function that were analyzed included left ventricular peak systolic pressure, end-diastolic pressure, first derivative of peak left ventricular pressure (dP/dt) and negative left ventricular dP/dt. Measurements were obtained during sinus rhythm, after paced premature ventricular depolarizations, during rapid ventricular pacing (cycle lengths 600 to 250 ms) and immediately after induction of sustained ventricular tachycardia. Mean left ventricular peak systolic blood pressure was 123 +/- 19 mm Hg during sinus rhythm, decreased to 77 +/- 23 mm Hg (p less than 0.05) at the induction of ventricular tachycardia and remained decreased during arrhythmia (p less than 0.01). Mean left ventricular end-diastolic pressure was 22 +/- 5 mm Hg during sinus rhythm, did not change after arrhythmia induction (22 +/- 9 mm Hg, p greater than 0.2) and remained unchanged during sustained ventricular tachycardia (p greater than 0.2). Mean peak left ventricular dP/dt was 1,400 +/- 620 mm Hg/s in sinus rhythm, decreased to 810 +/- 580 mm Hg/s (p less than 0.05) at ventricular tachycardia induction and remained decreased during sustained ventricular tachycardia (p less than 0.01).(ABSTRACT TRUNCATED AT 250 WORDS)

Clinical efficacy and electropharmacology of continuous intravenous amiodarone infusion and chronic oral amiodarone in refractory ventricular tachycardia.

The clinical efficacy and electropharmacologic effects of continuous intravenous (i.v.) amiodarone infusion (10 to 20 mg/kg/day for 4 to 7 days) followed by chronic oral amiodarone therapy (400 to 800 mg/day for 24 to 53 days) were evaluated in 17 patients with refractory sustained ventricular tachycardia (VT) or ventricular fibrillation. Intravenous amiodarone infusion prolonged the RR interval (from 754 +/- 85 to 860 +/- 157 ms, p less than 0.05), PR interval (from 192 +/- 53 to 212 +/- 54 ms, p less than 0.01) QRS duration (from 103 +/- 21 to 117 +/- 25 ms, p less than 0.001) and QTc interval (from 423 +/- 22 to 466 +/- 31 ms, p less than 0.001). Chronic oral amiodarone treatment had similar but more pronounced effects on electrocardiographic intervals. The ventricular effective refractory period tended to prolong after i.v. amiodarone infusion (p less than 0.1 to greater than 0.05) but prolonged significantly after chronic oral amiodarone (p = 0.025). Mean serum amiodarone concentration was 1.7 +/- 1.0 mg/liter with infusion and 1.5 +/- 0.6 mg/liter with oral therapy. Intravenous amiodarone infusion suppressed spontaneous VT in 5 of 9 patients with frequent VT recurrences, but had no effect on cycle length of spontaneous VT. Chronic amiodarone therapy either suppressed spontaneous VT recurrences or prolonged cycle length during VT recurrences. VT induction after i.v. amiodarone was not predictive of VT induction or spontaneous VT recurrences after chronic oral amiodarone treatment. Thus, i.v. amiodarone has limited value in acute control of VT and clinical or electrophysiologic response to it is not predictive of long term therapeutic results with amiodarone.

Chronic effects of amiodarone in patients with refractory ventricular tachycardia.

We examined the chronic electrophysiologic, systemic, and pharmacologic effects of chronic oral amiodarone therapy in 24 patients with refractory ventricular tachycardia and organic heart disease. Chronic amiodarone therapy resulted in significant increases in R-R interval (from 798 +/- 182 msec to 912 +/- 100 msec; P less than 0.01), P-R interval (from 205 +/- 66 msec to 221 +/- 87 msec; P less than 0.02), QRS duration (from 103 +/- 24 msec to 115 +/- 28 msec; P less than 0.001), and Q-Tc interval (from 413 +/- 48 msec to 470 +/- 46 msec; P less than 0.001). Significant increases were also noted in P-A interval (from 36 +/- 14 msec to 45 +/- 13 msec; P less than 0.05), A-H interval (from 119 +/- 61 msec to 141 +/- 87 msec; P less than 0.02), and H-V interval (from 52 +/- 12 msec to 64 +/- 11 msec; P less than 0.001). Electrophysiologic parameters showing changes included corrected sinus node recovery time (from 271 +/- 140 msec to 425 +/- 122 msec; P less than 0.01), the effective refractory period of the atrium (from 263 +/- 32 msec to 321 +/- 47 msec; P less than 0.01), the effective refractory period of the atrioventricular node (from 348 +/- 109 msec to 478 +/- 157 msec; P less than 0.001), the effective refractory period of the ventricle (from 253 +/- 21 msec to 291 +/- 28 msec; P less than 0.001), the atrial pacing cycle length producing A-V nodal Wenckebach (from 436 +/- 109 msec to 531 +/- 95 msec; P less than 0.001), and the functional refractory period of the A-V node (from 422 +/- 68 msec to 499 +/- 95 msec; P less than 0.001). Programmed electrical stimulation performed after 21-88 (mean 31) days of treatment was highly predictive of long-term results if suppression of arrhythmia induction was demonstrated (12 patients) or if the spontaneous arrhythmia was reinduced (5 patients). Induction of morphologically new ventricular tachyarrhythmias was frequent (42%) but had a low spontaneous recurrence rate (10%) during follow-up. Systemic parameters on chronic amiodarone therapy showed significant increases in total T4 and reverse T3, with no change in pulmonary function tests or left ventricular ejection fraction. Serum amiodarone levels at chronic electrophysiologic study ranged from 0.44-4.10 (mean 1.3) micrograms/ml. Long-term follow-up (2.5 to 20 months) demonstrated a marked improvement in clinical symptoms and mortality, but a significant recurrence rate of a well-tolerated slower arrhythmia persisted. Adverse effects on chronic amiodarone therapy were frequent (10 patients) and often disabling but required drug discontinuation in only 1 patient.