Transcription of a satellite DNA in the newt.

Satellite 2 is an abundant, 330-bp tandemly repeated sequence in the genome of the newt, Notophthalmus viridescens. This sequence is distributed throughout the genome on each of the 11 chromosomes. Both strands of satellite 2 are transcribed on the lampbrush chromosomes during oogenesis, probably as a result of readthrough from upstream structural gene promoters. In addition to these heterogeneous nuclear transcripts, satellite 2 is homologous to stable, strand-specific cytoplasmic transcripts in a variety of different tissues. The majority of these transcripts correspond in size to the entire satellite 2 repeat unit, or to whole multiples of the repeat. The transcripts present in the ovary have been sequenced by primer extension and were found to be more homogeneous than eight independently cloned satellite 2 DNA repeats. We propose that the stable cytoplasmic transcripts are encoded by a small subset of genomic satellite 2 sequences.

Mendelian and non-mendelian mutations affecting surface antigen expression in Paramecium tetraurelia.

A screening procedure was devised for the isolation of X-ray-induced mutations affecting the expression of the A immobilization antigen (i-antigen) in Paramecium tetraurelia. Two of the mutations isolated by this procedure proved to be in modifier genes. The two genes are unlinked to each other and unlinked to the structural A i-antigen gene. These are the first modifier genes identified in a Paramecium sp. that affect surface antigen expression. Another mutation was found to be a deletion of sequences just downstream from the A i-antigen gene. In cells carrying this mutation, the A i-antigen gene lies in close proximity to the end of a macronuclear chromosome. The expression of the A i-antigen is not affected in these cells, demonstrating that downstream sequences are not important for the regulation and expression of the A i-antigen gene. A stable cell line was also recovered which shows non-Mendelian inheritance of a macronuclear deletion of the A i-antigen gene. This mutant does not contain the gene in its macronucleus, but contains a complete copy of the gene in its micronucleus. In the cytoplasm of wild-type animals, the micronuclear gene is included in the developing macronucleus; in the cytoplasm of the mutant, the incorporation of the A i-antigen gene into the macronucleus is inhibited. This is the first evidence that a mechanism is available in ciliates to control the expression of a gene by regulating its incorporation into developing macronuclei.

Newt satellite 2 transcripts self-cleave by using an extended hammerhead structure.

Synthetic transcripts of satellite 2 DNA from newts undergo self-catalyzed, site-specific cleavage in vitro. Cleavage occurs within a domain that is similar to the hammerhead domain used by a number of self-cleaving, infectious plant RNAs. The newt hammerhead has a potentially unstable structure due to a stem composed of two base pairs and a 2-nucleotide loop, and unlike other hammerheads that have been studied, it cannot cleave as an isolated unit. Here we show that cleavage by a single newt hammerhead requires additional satellite 2 sequences flanking both ends of the hammerhead domain. We also present a structural model of a truncated satellite 2 transcript which is capable of cleavage. The structure includes an internally looped extension to one of the conserved stems of the hammerhead. By in vitro mutagenesis, the identities of each of the five nucleotides composing one of the internal loops were shown to be critical for cleavage. Additional evidence that the extension stimulates self-cleavage in a manner other than by simply stabilizing the hammerhead is presented.

Structure and expression of genes for surface proteins in Paramecium.

Surface proteins from 11 antigenic types of Paramecium tetraurelia vary in molecular weight from 251,000 to 308,000. The size of a series of polyadenylated RNAs obtained from these types were correlated with the sizes of the proteins and judged to be the mRNAs for the proteins. The mRNAs were used to identify genomic DNA clones containing complementary sequences. The gene for antigen A was present in one copy per genome, and the data suggest that extensive introns were absent. When restriction enzyme digests of DNA from cultures of paramecia with active and inactive genes were probed with portions of the cloned genes, no evidence for rearrangements or changes in gene dosage was found.

Two pathways of proton transfer reaction to (triphos)Cu(η(1)-BH4) via a dihydrogen bond [triphos = 1,1,1-tris(diphenylphosphinomethyl)ethane].

The interaction of the η(1)-tetrahydroborate copper(i) complex (triphos)Cu(η(1)-BH4) () with proton donors [CF3CH2OH (TFE), (CF3)2CHOH (HFIP), (CF3)3COH (PFTB), PhOH, p-NO2C6H4OH (PNP), p-NO2C6H4N[double bond, length as m-dash]NC6H4OH (PNAP), CF3OH] was a subject of a combined IR spectroscopic and theoretical investigation. Spectral (Δν) and thermodynamic (ΔH) parameters of dihydrogen bond (DHB) formation were determined experimentally. The terminal hydride ligand (characterized by the basicity factor Ej(BH) = 0.87 ± 0.01) is found to be a site of proton transfer which begins with nucleophilic substitution of BH4(-) by the alcohol oxygen atom on the copper center (BH pathway). The activation barrier computed for (CF3)2CHOH in CH2Cl2 - ΔG = 20.6 kcal mol(-1) - is in good agreement with the experimental value (ΔG = 20.0 kcal mol(-1)). An abnormal dependence of the reaction rate on the proton donor strength found experimentally in dichloromethane is explained computationally on the basis of the variation of the structural and energetic details of this process with the proton donor strength. In the second reaction mechanism found (CuH pathway), DHB complexes with the initial ROH coordination to the bridging hydride lead to B-Hbr bond cleavage with BH3 elimination. "Copper assistance" via the CuO interaction is not involved. This mechanism can be evoked to explain the occurrence of proton transfer in coordinating solvents.

Initial experience with larger laser sheaths for the removal of transvenous pacemaker and implantable defibrillator leads.

BACKGROUND: In a previous randomized trial, the 12F laser sheath removed pacing leads via the implant vein more successfully than traditional mechanical tools alone. Two larger sizes of laser sheath, the 14F and 16F models, were developed to extract defibrillator leads and large-diameter pacing leads implanted for the chronic. These devices use pulsed ultraviolet laser light to core though fibrotic tissue grown over the lead body to free the lead from the vasculature. A mandatory prospective registry studied the safety and effectiveness profiles of the larger laser sheaths vis-à-vis the 12F laser sheath. METHODS AND RESULTS: In this study, 863 patients underwent extraction of 1285 leads at 52 sites. Patients treated with the 14F device tended to have older leads than the 12F population; the 16F population, which comprised mostly defibrillator patients, were younger, had younger leads, and were more often male than the 12F population. Clinical success (extracting the entire lead or the lead body minus the distal electrode) was observed in 91% to 92% of cases for all device sizes. The overall complication rate was 3.6%, with 0.8% perioperative mortality. Incidence of complications was independent of laser sheath size. CONCLUSIONS: The 14F and 16F laser sheaths offer an extraction option for larger long-term transvenous pacemaker and defibrillator leads that is as safe and effective as the 12F laser sheath.

Radiofrequency catheter ablation in the treatment of supraventricular tachycardia in the elderly.

OBJECTIVES: The purpose of this study was to evaluate the efficacy and safety of radiofrequency catheter ablation for the treatment of supraventricular tachycardias in an elderly (> or = 70 years of age) group of patients. BACKGROUND: Supraventricular tachycardias are the most common form of cardiac arrhythmia and affect all age groups. Although usually well tolerated in youth, supraventricular tachycardias may be associated with disabling symptoms and have life-threatening potential in the elderly. In addition, antiarrhythmic agents are less well tolerated and may be associated with a higher incidence of toxicity in the elderly. METHODS: From May 1989 to March 1993, 454 patients underwent a radiofrequency catheter ablation procedure at the University of California, San Francisco, for the treatment of symptomatic supraventricular tachycardia. Sixty-seven of these patients were > or = 70 years of age and constituted the study group. Patients underwent one of the following catheter ablation procedures: complete atrioventricular (AV) junctional ablation for ventricular rate control in patients with atrial fibrillation (37 patients), AV node modification for the treatment of AV node reentrant tachycardia (17 patients), accessory pathway ablation (9 patients), ablation of the "slow zone" to cure atrial flutter (4 patients) and atrial tachycardia ablation (1 patient). One patient underwent ablation for both AV node reentrant tachycardia and atrial flutter. RESULTS: Success was achieved in 67 (98.5%) of 68 ablation procedures. There were no procedural or early deaths. The overall complication rate was 7.4%, and only one patient (1.5%) had long-term sequelae (permanent cardiac pacing for complete heart block). At a mean (+/- SD) follow-up of 22.1 +/- 12.9 months, 63 (94%) of 67 patients were alive, with no antiarrhythmic agents for the treatment of their presenting arrhythmia. CONCLUSIONS: In this series radiofrequency catheter ablation appears to be an effective and safe treatment option for elderly patients (> or = 70 years of age) with a variety of symptomatic, drug-resistant supraventricular tachycardias. Because of the high incidence of severe symptoms associated with tachycardic episodes, the expense and the possible severe proarrhythmic problems associated with antiarrhythmic medications in this age group, catheter ablation may be considered an early rather than a "last resort" treatment option.

Pacemaker lead extraction with the laser sheath: results of the pacing lead extraction with the excimer sheath (PLEXES) trial.

OBJECTIVES: The purpose of this study was to evaluate the safety and effectiveness of pacemaker lead extraction with the excimer sheath in comparison to nonlaser lead extraction. BACKGROUND: Fibrotic attachments that develop between chronically implanted pacemaker leads and to the venous, valvular and cardiac structures are the major obstacles to safe and consistent lead extraction. Locking stylets and telescoping sheaths produce a technically demanding but effective technique of mechanically disrupting the fibrosis. However, ultraviolet excimer laser light dissolves instead of tearing the tissue attachments. METHODS: A randomized trial of lead extraction was conducted in 301 patients with 465 chronically implanted pacemaker leads. The laser group patients had the leads removed with identical tools as the nonlaser group with the exception that the inner telescoping sheath was replaced with the 12-F excimer laser sheath. Success for both groups was defined as complete lead removal with the randomized therapy without complications. RESULTS: Complete lead removal rate was 94% in the laser group and 64% in the nonlaser group (p = 0.001). Failed nonlaser extraction was completed with the laser tools 88% of the time. The mean time to achieve a successful lead extraction was significantly reduced for patients randomized to the laser tools, 10.1 +/- 11.5 min compared with 12.9 +/- 19.2 min for patients randomized to nonlaser techniques (p < 0.04). Potentially life-threatening complications occurred in none of the nonlaser and three of the laser patients, including one death (p = NS). CONCLUSIONS: Laser-assisted pacemaker lead extraction has significant clinical advantages over extraction without laser tools and is associated with significant risks.

Use of P wave configuration during atrial tachycardia to predict site of origin.

OBJECTIVES: This study sought to construct an algorithm to differentiate left atrial from right atrial tachycardia foci on the basis of surface electrocardiograms (ECGs). BACKGROUND: Atrial tachycardia is an uncommon form of supraventricular tachycardia, often resistant to drug therapy. METHODS: A total of 31 consecutive patients with atrial tachycardia due to either abnormal automaticity or triggered rhythm underwent detailed atrial endocardial mapping and successful radiofrequency catheter ablation of a single atrial focus. P wave configuration was analyzed from 12-lead ECGs during tachycardia during either spontaneous or pharmacologically induced atrioventricular block. P waves inscribed above the isoelectric line (TP interval) were classified as positive, below as negative, above and below (or conversely, below and above) as biphasic and flat P waves as isoelectric (0). In 17 patients the tachycardia was located in the right atrium: crista terminalis (n = 4); right atrial appendage (n = 4); lateral wall (n = 4); posteroinferior right atrium (n = 3); tricuspid annulus (n = 1); and near the coronary sinus (n = 1). In 14 patients, atrial tachycardia was located in the left atrium: at the entrance of the right (n = 6) or left (n = 4) superior pulmonary veins; left inferior pulmonary vein (n = 1); inferior left atrium (n = 1); base of left atrial appendage (n = 1); and high lateral left atrium (n = 1). RESULTS: There were no differences in P wave vectors between sites at the right atrial lateral wall versus the right atrial appendage or between sites at the entrance of right versus left superior pulmonary veins. However, analysis of P wave configuration showed that leads aVL and V1 were most helpful in distinguishing right atrial from left atrial foci. The sensitivity and specificity of using a positive or biphasic P wave in lead aVL to predict a right atrial focus was 88% and 79%, respectively. The sensitivity and specificity of a positive P wave in lead V1 in predicting a left atrial focus was 93% and 88%, respectively. CONCLUSIONS: 1) Analyses of surface P wave configuration proved to be reasonably good in differentiating right atrial from left atrial tachycardia foci. 2) Leads II, III and aVF were helpful in providing clues for differentiating superior from inferior foci.

Intracardiac echocardiography during radiofrequency catheter ablation of cardiac arrhythmias in humans.

OBJECTIVES: The purpose of this study was to describe our preliminary experience using catheter-based intracardiac echocardiography as an adjunct to biplane fluoroscopy for guiding radiofrequency catheter ablation of atrial arrhythmias in the right side of the heart. BACKGROUND: Catheter ablation requires precise positioning and stable ablation electrode-endocardial contact. This procedure is currently guided by an analysis of intracardiac electrograms and fluoroscopy. However, the use of fluoroscopy does not allow the endocardium and certain anatomic landmarks to be identified and is associated with the hazards of radiation exposure. METHODS: Seventeen symptomatic patients were studied. A 10F 10-MHz intracardiac imaging catheter was used to visualize specific anatomic landmarks in the right atrium for directing the ablation electrode in 15 patients undergoing radiofrequency ablation of 19 arrhythmias and to assist with interatrial septal puncture in 3 patients. RESULTS: Continuous intracardiac imaging was performed for a mean +/- SD of 63.6 +/- 39.2 min and demonstrated distal electrode-endocardial tissue contact in 81 (60%) of 134 radiofrequency applications. Movement of the catheter was demonstrated during 36 (44%), microcavitations during 39 (48%) and thrombus during 15 (19%) of the 81 imaged applications. In 7 of 10 procedures for atrial flutter, successful ablation was directed at anatomic corridors in the right atrium visualized with intracardiac echocardiography. During ablation of atrial tachycardia, imaging identified abnormal atrial anatomy related to previous surgery and guided successful ablation of a reentrant tachycardia circulating around these anatomic obstacles. In two procedures for slow pathway modification of atrioventricular node reentrant tachycardia, intracardiac echocardiography confirmed catheter stability at the tricuspid annulus anterior to the coronary sinus. CONCLUSIONS: During catheter ablation, intracardiac echocardiography augments fluoroscopy by visualizing anatomic landmarks, ensuring stable endocardial contact and assisting in transseptal puncture. Ablation of typical atrial flutter can be successfully directed at anatomic corridors identified using intracardiac imaging.

Cloning and characterization of extended hammerheads from a diverse set of caudate amphibians.

Satellite 2 (sat2) elements from six species representing four caudate amphibian families were cloned and analyzed. Despite differences in the abundance of this element, the tandemly repeated genomic organization found for sat2 in earlier investigations was conserved in the newly investigated species. The ability of sat2 transcripts to catalyze their own site-specific self-cleavage using an extended hammerhead (HH) motif had also been conserved. This motif is similar to the HH found in a number of infectious plant RNAs, but has an abbreviated stem III region and an internally looped stem I extension that distinguish it from the prototypical HH. The extended HH analyzed in this survey can be further organized into two groups on the basis of shared sequences and structural details in the peripheral stem-loop structures. HH derived from the family Salamandridae constitute one group, whereas HH from all other salamanders that have been investigated belong to the other group. Although many of the natural variants did not cleave efficiently in vitro, examples of relatively active variants for each design were found. This survey has therefore identified two designs that are suitable starting models for the structural and functional analysis of the extended HH.

Tissue-specific permutations of self-cleaving newt satellite-2 transcripts.

Synthetic transcripts of satellite-2 (sat2) DNA from the newt undergo self-catalyzed, site-specific cleavage in vitro using the conserved hammerhead (HH) motif found in a number of infectious plant RNAs. We have analyzed sat2 transcripts from a variety of tissues to obtain evidence for the occurrence of self-cleavage in vivo. We found two distinct types of sat2 transcript populations. Monomeric transcripts in the tests and in somatic tissues such as the liver have ends that map to the in vitro self-cleavage site, and have end groups (5'-hydroxyls and phosphate blocked 3' ends) similar to those produced by self-cleavage. This is the first indication of HH-mediated self-cleavage occurring in animal cells. The monomeric sat2 transcripts found in the newt ovary have a different permutation of the basic DNA repeat sequence. These transcripts begin and end 47 nucleotides upstream from the in vitro self-cleavage site. Despite their apparent lack of utilization of the self-cleavage site, they have end groups which indicate that processing is involved in their formation.

Initial experience with a laser sheath to extract chronic transvenous implantable cardioverter-defibrillator leads.

This is the first study of the use of a laser sheath to remove chronic transvenous implantable cardioverter-defibrillator leads. A 16Fr laser sheath was used to extract 14 leads from 11 patients, 6 of whom had undergone failed previous extraction attempts using conventional tools.

Presence and significance of the left atrionodal connection during atrioventricular nodal reentrant tachycardia.

It has been suggested that the anatomic substrates of dual atrioventricular nodal pathways are likely to be the atrionodal connections. During atrioventricular nodal re-entrant tachycardia (AVNRT) or ventricular pacing (VP), an earliest retrograde atrial activation in the coronary sinus (CS) distal to the ostium (CS breakthrough) would suggest the presence of an exit from a left atrionodal connection. The aim of the study was to evaluate the incidence of such an atrial retrograde activation in the CS during AVNRT and VP. The retrograde atrial activation was recorded during typical AVNRT (38 patients, 27 women, mean age 44 +/- 18 years) by a multipolar catheter in the CS, a decapolar catheter in the His bundle position, and a deflectable quadripolar catheter along the tricuspid annulus anterior to the CS ostium. In 31 patients the retrograde atrial activation was recorded also during VP at a similar cycle length. A CS breakthrough was found in 18 patients during AVNRT (47%) and in 13 patients during VP (42%). Presence or absence of CS breakthrough was concordant between AVNRT and VP in 90% of the patients. A CS breakthrough, suggesting a left-sided atrionodal connection, is frequently recorded both during AVNRT and VP. In patients with a CS breakthrough pattern, the absence of correlation between the His bundle to the earliest CS retrograde atrial electrogram interval and AVNRT cycle length, or any other atrial activation times recorded in the posterior and anterior region of the Koch's triangle, would suggest that the left-sided atrionodal connection is a bystander during typical AVNRT.

Clinical and electrophysiologic features and role of catheter ablation techniques in adult patients with automatic atrioventricular junctional tachycardia.

A total of 8 patients with junctional tachycardia (JT) were included for study. Patients with JT had a supraventricular arrhythmia that was initiated by a junctional complex without PR prolongation and episodes of atrioventricular (AV) dissociation. JT could not be initiated by pacing and occurred either spontaneously (3 patients) or with isoproterenol (5 patients). Tachycardia could be consistently terminated by either carotid sinus massage (1 patient), intravenous adenosine (2 patients), or critically timed ventricular premature complexes (3 patients). In 6 of the 8 other patients, tachycardia foci (atrial or ventricular) or mechanisms (AV node reentry) were found. Two patients underwent complete AV junctional ablation and 2 had termination of tachycardia without change in the AV conduction by perinodal application of radiofrequency lesions. AVJT appears to be due to abnormal automaticity and may be successfully ablated by application of radiofrequency energy to perinodal areas.

Ventricular septal summit stimulation in atrioventricular nodal reentrant tachycardia.

In all, 18 consecutive patients with atrioventricular nodal reentry tachycardia (AVNRT) underwent right ventricular (RV) stimulation during AVNRT from either the RV apex or summit. Stimulation from the RV apex advanced the tachycardia with the same atrial sequence in 6 of 18 patients (33%), but never conclusively excluded the presence of a low atrial tachycardia. RV summit stimulation resulted in direct stimulation of the low septal right atrium in 6 patients. RV summit stimulation advanced the tachycardia in 4 patients, delayed it in 2 and terminated it in 3 without an atrial electrogram. The latter 2 findings exclude the presence of a low atrial tachycardia. Thus, in patients with AVNRT, application of extrastimuli closer to the putative reentrant site enables greater efficacy in tachycardia resetting and in excluding a low septal atrial tachycardia.

Evaluation of outpatient initiation of antiarrhythmic drug therapy in patients reverting to sinus rhythm after an episode of atrial fibrillation.

The safety of a method of outpatient antiarrhythmic drug loading utilizing a continuous loop event recorder was evaluated. The findings suggest that the standard 2-day hospital admission for drug loading is not necessary in all patients and a method of outpatient loading may be equally safe.

Campylobacter jejuni infections in Haifa subdistrict, Israel, summer 1981.

A study investigating the clinical and epidemiological characteristics of Campylobacter jejuni infections in an urban community in Israel is presented. Most of the information was obtained by interviewing 76 patients who constituted a systematic sample out of a total of 215 patients suffering from acute Campylobacter jejuni diarrhoea during the summer of 1981. The crude annual incidence rate was 17 per 10 000. Age-specific incidence rate in infants up to one year of age was eight times higher than that in children 1-14 years of age. The median duration of infection until a negative culture was obtained in convalescent patients, was 10 days. No evidence of resistance to erythromycin was found. No statistically significant difference in keeping animals at home was found between the patients and a neighbourhood control group. The frequencies of the various complaints and clinical findings are described.

Effect of patient characteristics on the yield of prolonged baseline head-up tilt testing and the additional yield of drug provocation.

OBJECTIVE: To define the value of tilt testing and hte additional yield of drug provocation over prolonged baseline tilt in different patient subgroups. (Many different protocols are in use for head-up tilt testing in heterogeneous groups of patients. Not all patients in reported series have recurrent syncope, and there is often a wide age range and a variable incidence of structural heart disease.) DESIGN: In a prospective study, baseline 60 degrees head-up tilt testing was undertaken for 45 minutes, initially without drug provocation. Patients who remained symptom free were given intravenous isoprenaline (isoproterenol) and further tilting or edrophonium (10 mg bolus) during tilt, in an order determined randomly before the start of the test. If they were symptom free after the first drug, they were given the other drug. A positive test was recorded when syncope or pre-syncope occurred with a rapid fall (> 30%) in blood pressure. The impact on tilt result of the type of symptoms, presence of significant structural heart disease (SHD), presence of a non-cardiovascular cause of sudden diminished consciousness (SDC), and age was then assessed by subgroup analysis. PATIENTS: 145 patients (73 female, mean age 51 (25), range 8-94) with one or more episodes of pre-syncope or syncope. RESULTS: 39 patients (27%, 21 female, age 49 (25) years) had positive tests and 106 (73%, 52 female, age 52 (25) years) negative tests. 27 (69%) had a positive test during baseline tilt at 20.5 (10.8) minutes, five (13%) with isoprenaline infusion, and seven (18%) with edrophonium bolus. Patients with recurrent syncope rather than single syncopal episodes or single or recurrent pre-syncope were more likely to have a positive tilt test (41% v 17%, P < 0.005) and patients with SHD or SDC (69/14 patients) were much less likely than patients without (16% v 42%, P < 0.0001). The yield of positive tests was similar if patients were below (26%) or above (27%) the mean age (50 years). When multiple factors were combined, the yield ranged from 0% for 21 patients under 50 years with SHD or SDC and without recurrent syncope to 73% in 11 patients over 50 years with recurrent syncope and no SHD or SDC. The additional yield in subgroups over 45 minute baseline tilt (70 (11)%) of isoprenaline (13 (10)%) was similar to that of edrophonium (17 (8)%, P = NS), but six (50% of those who were drug positive) patients required a second drug to produce a positive result (two with isoprenaline second, four with edrophonium second). CONCLUSIONS: Head-up tilt testing in a heterogeneous population has a low yield. Simple clinical characteristics define the type of patient who is likely to have a positive tilt test and the patient who is not and in whom other investigations should receive priority. The great majority of positive tests will occur during prolonged baseline testing if this is used. Isoprenaline and edrophonium produced similar additional yields of positive tests.

An evaluation of an education programme, for prevention and early diagnosis of malignancy in Israel.

A community-wide health education programme for the prevention and early detection of cancer was performed in the Krayot municipalities, north of Haifa, Israel, during the years 1985-86. In order to assess its effectiveness, an evaluation programme was conducted. The knowledge, attitudes and behaviour of women who had been exposed to the education programme and of a control group, were compared before the programme started and 2.5 years later, after it ended. The programme was conducted in five clinics and included changes in the functioning of the clinics and their teams to be used in the interaction with the insured women. A piloted questionnaire was used to evaluate knowledge, attitudes and behaviour of these women. Emphasis was placed on smoking, sun tanning, self-examination of breast and other aspects of possible early detection of cancer. The education programme was ineffective not only with regard to behaviour but also in relation to knowledge on the relationship between health behaviour and cancer. The need for the establishment of more effective means for health education is eminent. If health education programmes are to succeed, we must understand what motivates or prevents compliance with recommended health behaviour.