Unusual thionation of a cyclic hexapeptide

One carbonyl oxygen of the cyclic hexapeptide cycle(-Gly¹-Pro²-Phe³-Val⁴-Phe⁵-Phe⁶-) (A) can be selectively exchanged with sulphur using Yokoyama's reagent. Surprisingly it was not the C=O of Gly¹ but that of Phe⁵ which was substituted and cyclo(-Gly¹-Pro62-Phe³-Va1⁴-Phe⁵ψ[CS-NH]Phe⁶-) (B)was obtained. Thionation results in a conformational change of the peptide backbone although the C=O of Phe⁵ and the corresponding C=S are not involved in internal hydrogen bonds. Two isomers in slow exchange, containing a CIS Gly¹-Pro² bond in a βVIa-turn (minor) and a trans Gly-Pro bond in a βII′-turn (major), were analyzed by restrained molecular dynamics in vacuo and in DMSO as well as using time dependent distance constraints. It is impossible to fit all experimental data to a static structure of each isomer. Interpreting the conflicting NOES, local segment flexibility is found. MD simulations lead to a dynamic model for each structure with evidence of an equilibrium between a βI- and βII-turn about the Val⁴-Phe⁵ amide bond in both the cis and trans isomers. Additionally proton relaxation rates in the rotating frame (R_1p) were measured to verify the assumption of this fast βI/βII equilibrium within each isomer. Significant contributions to R_1p-rates from intramolecular motions were found for both isomers. Therefore it is possible to distinguish between at least four conformers interconverting on different time scales based on NMR data and MD refinement. This work shows that thionation is a useful modification of peptides for conformation-activity investigations.     

Experimental Evaluation of the Dialdehyde Starch Preserved Bovine Internal Mammary Artery as a Small Diameter Arterial Substitute

A canine femoral artery model was used for evaluation of a dialdehyde starch preserved bovine internal mammary artery (BIMA) (3 and 4 mm internal diameter) in comparison to a polytetrafluoroethylene (PTFE) graft. The study comprised three groups for a 2-hour (n = 7, 3 mm), 3-month (n = 10, 4 mm), and 6-month (n = 10, 4 mm) follow-up. The thrombogenicity of the grafts was measured after 2 hours and 3 months using chromium 51 labeled autologous platelets. In addition, compliance studies were done. To control the wall stability of the xenografts, the collagen content before implantation and after explantation was examined together with the diameter of the grafts. Healing characteristics were studied using appropriate histologic methods. The acute platelet adhesion rate (2 hours) of the BIMA graft was 181 +/- 69 x 10(4) platelets/mm2 as compared to 57 +/- 43 x 10(4) for PTFE (p less than 0.05, t-test). However, after 3 months the thrombogenicity of the biograft had decreased whereas the platelet count at the PTFE graft had increased (BIMA: 79 +/- 48 x 10(4); PTFE: 179 +/- 102 x 10(4), p less than 0.05). At implantation, the compliance of the BIMA graft was 0.028% +/- 0.009% per mmHg as compared to 0.06% +/- 0.0025% of the femoral artery. The PTFE graft was uncompliant (0.008 +/- 0.005). After 6 months, the compliance of the femoral artery had decreased to 0.039% +/- 0.013% per mmHg, which was now well matched to the nearly unchanged compliance of the biograft (0.0027 +/- 0.005).(ABSTRACT TRUNCATED AT 250 WORDS)     

High Resolution ECG and Left Ventricular Volume After Acute Myocardial Infarction

The presence of ventricular late potentials (LPs) early after acute myocardial infarction (AMI) was recently reported to correlate with left ventricular dilatation subsequent to AMI. We assessed prospectively the relationship between LP (time domain) in the late phase of AMI and left ventricular end-diastolic volume (EDV) measured by equilibrium radionuclide angiocardiography 4 weeks and 12 months after AMI. In 80 consecutive patients 4 weeks and 12 months after thrombolytic therapy for AMI, LP and EDV were deter mined (EDV1, EDV2). There was no significant correlation between QRS duration (r = 0.18), RMS40 (r = 0.08), or LAS40 (r = 0.1) and EDV1 or EDV2 in patients with or without LP at baseline. In both groups (patients with [n = 15] and without LP [n = 65]), EDVl and EDV2 were comparable (128 ± 32 mL vs 126 ± 35 mL; 114 ± 40 mL vs 117 ± 36 mL; P = NS). In addition, there was no significant difference between EDVl and EDV2 in patients who developed new LP (n = 6) or lost LP (n = 9) 12 months after AMI. In contrast to LP in the very early phase after AMI, there seems to be no significant correlation between the high resolution ECG in the late phase after thrombolytic therapy for AMI and left ventricular EDV.     

A Fractally Coated, 1.3 mm2 High Impedance Pacing Electrode

Minimizing the geometric surface area of pacing electrodes increases impedance and reduces the current drain during stimulation, provided that voltage (pulse-width) thresholds remain unchanged. This may be feasible by coating the electrode surface to increase the capacity of the electrode tissue interface and to diminish polarization. Ten unipolar, tined leads with a surface area of 1.3 mm² and a “fractal” coating of Iridium (Biotronik SD-V137) were implanted in the ventricle, and electrogram amplitude (unfiltered), slew-rate, pacing threshold (0.5 ms), and impedance (2.5 V; 0.5 ms) were measured by the 5311 PSA (Medtronic). On days 0, 2, 5, 10, 28, 90, 180, 360 postimplant, sensing threshold (up to 7.0 mV, measuring range 1–14 mV on day 360 only) and the strength duration curve (0.5–4.0 V; 0.03–1.5 ms; steps: 0.5 V; 0.01 ms, respectively) were determined, the minimum charge delivered per pulse (charge threshold), and the impedance were taken from pacemaker telemetry (Intermedics 294–03). Data were compared with those of an earlier series of 20 unipolar, tined TIR-leads (Biotronik) with a surface area of 10 mm² and a “fractal” coating of titanium nitride. With the model SD-V137 versus TIR, intraoperative electrogram amplitudes were 15.1 ± 6.1 versus 14.4 ± 3.9 mV(NS), slew rates 3.45 ± 1.57 versus 1.94 ± 1.06 V/s (P < 0. 05), pacing thresholds 0.16 ± 0.05 versus 0.52 ± 0.15 V (P < 0.01) and impedance measurements 1,136 ±175 versus 441 ± 73 Ω (P < 0.0001), respectively. During follow-up, sensing thresholds were the same with both leads. Differences in pulse width thresholds lost its significance on day 28 but resumed on day 360 (SD-V137: 0.08 ± 0.04 ms; TIR: 0.16 ± 0.06 ms at 2.5 V; P < 0.01). With an electrode surface of 1.3 mm², charge per pulse and impedance consistently differed from control, beingO.15 ± 0.15 versus 0. 66 ± 0. 20 μC (P < 0.001) and 1,344 ± 376 versus 538 ± 79 Ω, respectively, one year after implantation (P < 0.0001). In summary, “fractally” coated small surface electrodes do not compromise sensing; by more than doubling impedance against controls they offer pacing thresholds (mainly in terms of charge) that are significantly lower than with the reference electrode.     

Mapping Guided Laser Catheter Ablation of the Atrioventricular Conduction in Dogs

Safety and efficacy of mapping guided laser catheter ablation of the AV junction was tested in a canine model. A total of 43 laser pulses (continuous wave, Nd:YAG, 1,064 nm, 30 W, irradiated spot diameter 2.0–2.5 mm) were delivered in 15 dogs (2–5 per dog) via a novel laser catheter system. Pulses were selectively aimed at: (1) the AV node: (2) the His bundle; and (3) the bundle branches. Laser pulses of 9.7 ± 1.1 seconds (n = 31) produced reversible conduction disturbances in the targeted segment of the AV conduction system, while pulses of 28.6 ± 7.9 seconds (n = 9) resulted in chronic block. The dogs survived the procedure without complications. Follow-up was 6.5–10.5 months. Histopathologically, lesions showed clear-cut oval-shaped areas of fibrosis of 0.5–18.0 mm in diameter and 0.5–3.5 mm (transmural) in depth, depending on the irradiation time. Pervenous mapping guided laser catheter irradiation of the AV junction can produce AV block consistently and selectively in the targeted segment of the right ventricular conduction system in dogs. The method is safe and can be performed in a controllable manner by using the catheter system presented.     

ORAL PHENTOLAMINE AS TREATMENT FOR ERECTILE DYSFUNCTION

Purpose

For most patients with erectile dysfunction oral agents are a preferred treatment option. Oral or buccal phentolamine has been shown to produce full erections in impotent subsets of study populations. We evaluate the efficacy of oral phentolamine.

Materials and Methods

After a comprehensive evaluation 44 patients with recent onset (less than 3 years) of erectile dysfunction and a high likelihood of organogenic etiology underwent a prospective, double-blind and placebo controlled trial with oral phentolamine after placebo.

Results

After placebo 4 of the 44 patients who reported full erections were excluded from study. Of the 40 patients in the double-blind phase full erections were achieved by 2 of 10 with placebo, and 3 of 10 with 20 mg., 5 of 10 with 40 mg. and 4 of 10 with 60 mg. phentolamine. There were no serious complications observed during the study, and only a single minor side effect occurred in 1 patient after 60 mg. phentolamine.

Conclusions

Our results indicate that oral phentolamine may be of benefit for the treatment of erectile dysfunction. Further studies are required to corroborate our findings.     

Clinical Significance of Sleep-Related Breathing Disorders in Patients with Implantable Cardioverter Defibrillators

The prevalence and clinical significance of sleep-related breathing disorders (SRBDs) in patients with cardiac disease and a history of life-threatening ventricular tachyarrhythmias is unclear. Forty consecutive recipients of implantable cardioverter defibrillators (ICDs) with cardiac disease and a documented history of spontaneous, life-threatening, ventricular tachyarrhythmias underwent full night polysomnography. SRBDs were diagnosed if the apnea/hypopnea index was > 10. SRBD were diagnosed in 16 of 40 patients (40%): central sleep apnea (CSA) was present in 9 of these 16 patients (56%), 8 of whom had associated Cheyne-Stoke respiration. Seven of the 16 patients with SRBD (44%) had obstructive sleep apnea (OSA). Patients with and without SRBDs were comparable with respect to left ventricular ejection fraction, NYHA classification, underlying heart disease, ICD indications, and concomitant antiarrhythmic drug and beta-blocker therapy. Patients were followed prospectively for 2 years. ICD-treated ventricular tachyarrhythmias occurred in 10 of 24 patients (42%) without SRBD, in 4 of 9 patients (44%) with CSA, and in 3 of 7 patients (44%) with OSA (NS). The numbers and circadian distributions of episodes recorded during follow-up in patients without SRBD versus with CSA or OSA were not significantly different (14 ± 25, median = 4 vs 4 ± 5, median = 2.5 vs 15 ± 15, median = 7, respectively). The 2-year mortality, which was entirely attributable to nonsudden cardiac events, was highest in patients with CSA (4/9 [44%], vs 0/7 [0%] with OSA, vs 3/24 patients (12.5%) without SRBD; P < 0.05).     

Determinants of the Natural Course of Ventricular Late Potentials After Thrombolytic Therapy for Acute Myocardial Infarction

The intraindividual changes of ventricular late potentials and their possible determinants were examined prospectively in 88 consecutive patients (male: 75; mean age: 58 ± 9 years) after thrombolytic therapy for acute myocardial infarction. Late potential analysis was performed 4 weeks and 12 months after acute myocardial infarction. At the same time, a left heart catheterization was performed to assess the extent of coronary heart disease and left ventricular ejection fraction. The incidence of late potential 4 weeks after acute myocardial infarction was 15% (13/88 patients). Eighteen percent (16/88) of the patients revealed changing results of late potential analysis: 9 patients lost late potential (late potential pos./neg.) 1 year after acute myocardial infarction and 7 patients presented new formation of late potential (late potential neg./pos.). Preserved late potentials were found in four patients (late potential pos./pos.). Late potential analysis remained negative in 68 patients (late potential neg./neg.). There was no influence of age, gender, site of infarction, clinical course, and medical treatment on the natural course of late potential. Changing results of late potential analysis seemed to be correlated with the evolution of left ventricular ejection fraction and the dynamics of coronary heart disease. In the group late potential pos./pos., comparable values for left ventricular ejection fraction were measured at both examinations, whereas late potential neg./neg. had a significant increase in ejection fraction. In the group late potential pos./neg., a significant improvement in left ventricular function was also measured. In contrast, the late potential neg./pos. group tended to have lower left ventricular ejection fractions 1 year after infarction. In the late potential neg./pos. and late potential pos./pos. groups, the extent of coronary artery disease returned to conditions comparable to baseline despite an initial reduction after coronary revascularization performed 4 weeks after infarction. Late potential neg./neg. and late potential pos./neg. revealed a stable benefit gained from coronary revascularization with a persistent reduction in the number of diseased vessels. Dynamic changes in the results of the signal-averaged ECG 1 year after thrombolytic therapy for acute myocardial infarction were observed in 18% of the patients. These changes seem to be correlated with the evolution of left ventricular function and the dynamics of coronary artery disease.     

THE EFFECT OF THE SPECIFIC PHOSPHODIESTERASE (PDE) INHIBITORS ON HUMAN AND RABBIT CAVERNOUS TISSUE IN VITRO AND IN VIVO

Purpose

Phosphodiesterases (PDE) are key enzymes in the regulation of the smooth muscle tone. Experimental studies showed PDE III and V-isoenzymes to play an important role in the smooth muscle tone regulation of corpus cavernosum. Recently, a specific PDE III-inhibitor (milrinone) and a PDE V-inhibitor (sildenafil) were introduced in clinical studies. An experimental study was done to examine a potential role of PDE-inhibitors in the treatment of erectile dysfunction.

Materials and Methods

In the organ bath, strips from human and rabbit corpus cavernosum were precontracted and increasing doses of PDE inhibitors were added. In patients with erectile dysfunction as well as in rabbits, intracavernous injections of milrinone were done.

Results

PDE-inhibitors dose-dependently relaxed human and rabbit corpus cavernosum strips. In the precontracted human cavernous tissue, milrinone and sildenafil were equally potent and efficacious in vitro. In the rabbit, milrinone induced slight tumescence but dramatic circulatory side effects. In patients, penile tumescences as well as full erections were observed.

Conclusions

Milrinone strongly relaxes human cavernous smooth muscle cells but it exhibits low relaxant effects in the rabbit cavernous tissue. In human tissue, sildenafil was equieffective with milrinone in vitro. In vivo, milrinone induced a good erectile response in humans but a poor erectile effect in rabbits. Our results support a possible potential for selective PDE-III and -V inhibitors in the treatment of impotence and give further evidence that the rabbit is an animal model of limited value to study the effects of drugs on cavernous smooth muscle tone regulation in vivo.     

Analysis during Sinus Rhythm and Ventricular Pacing of Reentry Circuit Isthmus Sites in Right Ventricular Cardiomyopathy

Background: The entrainment mapping algorithm is used for ablation of ventricular tachycardia (VT) in right ventricular (RV) cardiomyopathy, but ablation at endocardial isthmus sites has only a moderate success rate. This study was performed to identify additional local electrogram characteristics associated with successful ablation. Patients and Methods: Using entrainment mapping, 45 reentry circuit isthmus sites were detected in 11 patients with RV cardiomyopathy presenting with 13 monomorphic VTs. Local bipolar electrograms were retrospectively analyzed at reentry circuit isthmus sites during VT, sinus rhythm, and programmed stimulation from the right ventricular apex (RVA), and compared between successful and unsuccessful ablation sites. Results: Ablation was successful at 10 reentry circuit isthmus sites and unsuccessful at 35 isthmus sites. During VT, a longer endocardial activation time relative to QRS onset, an increased electrogram‐QRS interval as a percentage of VT cycle length, and a longer electrogram duration were found at successful in comparison to unsuccessful ablation sites. The presence of isolated diastolic potentials during sinus rhythm at reentry circuit isthmus sites, consistent with slow conduction or unidirectional conduction block, was associated with successful catheter ablation. Prolongation of the duration of the local multipotential electrogram by >100 ms during programmed RVA pacing at reentry circuit exit sites, indicating functional conduction disorder was also a marker of successful ablation. Conclusions: The demonstration of multipotential electrogram characteristics indicating fixed or functional conduction block may increase the likelihood of successful VT ablation at exit and central isthmus sites of reentry circuits in RV cardiomyopathy.