Oncogene-mediated propagation of tracheal epithelial cells from two cystic fibrosis fetuses with different mutations. Characterization of CFT-1 and CFT-2 cells in culture

Abstract. Primary tracheal epithelial cells obtained from two fetuses with cystic fibrosis (CF) were successfully transfected with a plasmid vector recombined with the large T oncogene of SV40. The resulting tracheal cells were propagated in culture for up to 25 passages and retained the mutations of the CF genes carried by the two fetuses, one heterozygous for the S549N and N1303K substitutions (CFT-I cells), and the other homozygous for the most common deletion ΔF508 (CFT-2 cells). The transfected cells: (a) expressed the SV40 large T oncogene, as determined by immunofluorescence and Northern blot analysis; (b) retained typical epithelial morphology, as assessed by the presence of microvilli, desmosomes, gap junctions, and cytokeratin expression; (c) were fully responsive to the cAMP-stimulating agents isproterenol, forskolin and vasoactive intestinal peptide for cAMP production and PKA activation; (d) do not produce any tumour in the athymic nude mice; (e) were diploid and tetraploid with a normal chromosomal complement at early passages, and (f) exhibited the abnormal regulation of chloride conductance characteristic of CF.
These results indicate that CFT-1 and CFT-2 cells constitute a suitable model for: (a) comparison of the maturation and function of the CFTR protein mutated in the two nucleotide-binding domains; (2) analysis of the biochemical defect in CF epithelial airway cells, (c) development of new therapeutic agents, and correction of the CF defect by gene replacement therapy in vitro .     

Hemodynamic Deterioration Following Radiofrequency Ablation of the Atrioventricular Conduction System

Radiofrequeucy ablation of the atrioventricular conduction system (ACS) has become an estoblished theTapy for patients with drug refroctory atrial fibrillation. We observed eight patients with hemodynamic deterioration ofteT radiofrequency oblotion of the otTioventTicular conduction system. As we found hemodynamic deterioration related to worsening mitral regurgitation, we compared the clinical history, eiectrophysiologicai, ond echocardiographic dato from the patients with hemodynamic deteriorotion and worsening mitral regurgitation (group 1) to those without hemodynamic deterioration and stable mitral regurgitation after the procedure (group 2). Eight out of 108 patients (7.4%) undergoing ablation of the ACS deteriorated hemodynamically with acute pulmonary edema in three and congestive heart failure in five patients occurring at a mean of 3 and 8 weeks, respectively, after the procedure. Three of these patients were referred for mitral valve surgery. Two patients underwent ablation using a left-sided approach. A right-sided approach was used in five patients. In one patient, a left- and right-sided approach was used. Compared to group 2 patients, group 1 patients had significantly higher left ventricular end-diastolic diameters (64 ± 6 mm vs 56 ± 9 mm) at baseline despite similar fractional shortening (32%± Il% vs 34%± 13%), left ventricular end-systolic diameters (43 ± 9 mm vs36 ± 7 mm) and degree of mitral regurgitation (1.4 ± 1.1 vs 1.4 ±0.7) on echocardiographic analysis. Thus, hemodynamic deterioration together with progression of mitral regurgitation is a potential complication of ablation of the ACS (up to 7.4%). Patients with high left ventricular end-diastolic diameters ond moderate mitral regurgitation at baseline seem prone to this complication.     

Intravascular Extraction of Chronic Pacemaker Leads: Efficacy and Follow-Up

Extraction of chronic pacemaker leads has been recommended for infections, prevention of venous thrombosis, migration, and possible perforation. Success with constant traction techniques has been variable, and the cost and morbidity of open chest surgical procedures are prohibitive. Efficacy of a new system for lead extraction using intravascular techniques was analyzed. The system (Cook Pacemaker) uses a locking stylet, which is secured at the distal electrode by counterclockwise rotation to reinforce the lead and facilitate traction, and dilator sheaths that are used to free the lead from adhesions in the venous system. In a series of 56 patients (ages 19–88)who presented for lead extraction because of erosion (5), infection (14), lead replacement (35), or other (2), 86 leads were extracted. Thirty-two were atrial leads and 54 ventricular; 23 had active fixation and 63 passive. Average duration of implant was 58 ±42 months (range 1–264). Eighty-four leads were totally removed and two partially removed. For these two leads, the distal tip was not removed; in both cases the locking stylet was not secured at the distal electrode due to obstruction within the lead. Two patients developed arm edema following the procedure, which resolved with elevation. One patient developed a subclavian thrombosis, which resolved with warfarin anticoagulation. Four patients have expired due to unrelated causes. In conclusion, this intravascular approach for extraction of chronic leads is effective, and the procedure is safe when performed by experienced personnel.     

Comparison of Biphasic and Monophasic Pulses: Does the Advantage of Biphasic Shocks Depend on the Waveshape?

With present implantable defibrillators, the ability to vary the defibrillation technique has been shown to increase the number of patients suitable for transvenous system. As newer waveforms become available, the need for a flexible device may change. In addition, although it has been shown that the option of biphasic waveform may increase the defibrillation efficacy, this may depend upon the shape of the biphasic waveform used. Thirty patients undergoing transvenous defibrillator implant were included in the study. In 20 patients (group I), defibrillation efficacy of simultaneous monophasic, sequential monophasic, and biphasic waveform with 50% tilt was determined randomly. Similarly, in ten patients (group II) testing of simultaneous monophasic shocks and biphasic waveforms with 65% and 80% tilt was performed in random order. The electrode system used consisted of two transvenous leads and a subcutaneous patch in all 30 patients. In group I, 50% tilt biphasic waveform consistently provided similar or better defibrillation efficacy compared to monophasic waveforms (biphasic 7.5 ±5.1 joules vs simultaneous 17 ± 7.8 joules, P < 0.01; and vs sequential 17 ± 8.4 joules, P <0.01). In group II, 65% tilt biphasic pulse required less energy for defibrillation as compared with simultaneous monophasic shocks (9.6 ± 4.5 joulesvs 15.6 ± 5.1 joules, P = 0.04). No significant difference was observed in terms of defibrillation threshold between 80% tilt biphasic shocks and simultaneous monophasic pulses (11.8 ± 6 joules vs 15.6 ±5.1 joules, P = NS). Biphasic shocks with smaller tilt delivered using a triple lead system more uniformly improved defibrillation threshold over standard monophasic waveforms.     

Diabetic ketoacidosis

Diabetic ketoacidosis is a common medical emergency and may represent up to 14% of all diabetic admissions with an annual incidence of about 4.6 cases per 1,000 diabetics per year. A spectrum of disease is encountered and mortality rates vary in different series from 1.9% to 15%. The pathogenesis of diabetic ketoacidosis is discussed in detail with particular emphasis on interpretation of biochemistry results and on fluid and electrolyte resuscitation. A flexible approach to fluid resuscitation, careful interpretation of biochemistry results, and close monitoring should be combined with other standard aspects of management such as titratable insulin infusions and early treatment of identifiable precipitants in order to minimise morbidity and mortality.     

Epicardial Mapping: Ventricular Epicardial Activation is Unaffected by Heart Position

Epicardial ventricular mapping was performed in five dogs during sinus rhythm with a sock array containing 41 bipolar electrodes. Maps were generated with a computer-assisted mapping system when the heart was in situ and when the heart was lifted by 44 degrees out of the chest. Times of earliest and latest epicardial activation in these two states did not differ. Despite a different frontal plane QRS axis, location of earliest activation was not affected by lifting the heart. In two of the five animals, the site of latest epicardial activation was minimally different from the heart in situ, but the general pattern of epicardial activation was unchanged. Therefore, the change in frontal plane QRS axis with lifting the heart was due to a change in heart position rather than a general change of heart activation.     

Clinical Efficacy and Safety of the New Cardioverter Defibrillator Systems

Clinical efficacy and safety of two new third-generation implantable cardioverter defibrillators (ICD) were studied in 38 patients with ventricular tachycardia (VT) or fibrillation (VF). There were 31 patients with coronary disease, three patients with right ventricular dysplasia, one patient with dilated cardiomyopathy, and three patients with valvular disease. Twenty-four patients (group I) received an ICD with monophasic (Ventak PRx 1700, CPI) and 14 patients (group II) with biphasic shocks (Cadence V 100, Ventritex). Intraoperatively, the mean defibrillation threshold was significantly lower in group II than in group I, both in patients with induced VT (group I 11.0 ± 6.3 joules: group N 5.8 ± 1.3 joules) (P < O.01) and induced VF (group I 17.5 ± 4.6 joules; group II 9.6 ± 5.2 joules) (P < O.O1). During the mean follow-up of 12 ± 7 months four patients (11%) died. 865 arrhythmia events (AE) occurred and were terminated by ATP (671 VTs, 78%). Acceleration of VTs was observed in 28 AE (3%) and ATP was unable to interrupt 58 AE (7%). ICD shocks were delivered as a first therapy in 108 AE (13%).     

Transcoronary Ethanol Ablation of the Atrioventricular Node in a Young Patient with Tricuspid Atresia

Catheter ablation of AV conduction with radiofrequency energy can be challenging in the presence of structural abnormalities of the AV junction, either congenitally or after reconstructive surgery. We used transcoronary ethanol to ablate the AV node in a patient with classic tricuspid atresia and refractory intraatrial reentry tachycardia. This approach provides an alternative means of creating complete heart block with catheter-based techniques, when radiofrequency catheter ablation is technically impossible or ineffective.     

An Absolute Requirement for P-Selectin in Ischemia/Reperfusion-Induced Leukocyte Recruitment in Cremaster Muscle

Objective : To systematically examine a role for P-selectin in a model of striated muscle ischemia/reperfusion (I/R). Methods : Ischemia was induced in the cremaster muscle of mice by occluding the main feeding arteriole for 30 minutes. Blood flow was then restored to allow for 60 minutes of reperfusion and leukocyte kinetics were assessed during the control period (before I/R) and at 5, 30, and 60 minutes of reperfusion. To study a role for P-selectin in this model, three different approaches were used: Wildtype animals received fucoidin (10 mg/kg, iv), an anti-P-selectin antibody (RB40.34; 20 mg/animal, iv) at 25 minutes of ischemia, or I/R was induced in P-selectin-deficient mice. Results : Ischemia/reperfusion induced a rapid and significant increase in leukocyte rolling, adhesion, and emigration in wild-type mice. The I/R-induced increase in leukocyte rolling was transient, inasmuch as it was reduced by approximately 50% at 30 minutes of reperfusion, and returned to control levels by 60 minutes. Both fucoidin and an anti-P-selectin antibody completely prevented the I/R-induced increase in leukocyte rolling. The P-selectin-deficient animals exhibited absolutely no baseline leukocyte rolling, adhesion, or emigration. Furthermore, I/R did not induce any increase in leukocyte recruitment in the Pselectin-deficient animals over the first 60 minutes of reperfusion. Conclusion : The results from this study clearly illustrate that P-selectin is absolutely critical in both baseline and I/R-induced leukocyte infiltration in the murine-striated muscle.