Evaluation of the Safety and Efficacy of Deep Sedation for Electrophysiology Procedures Administered in the Absence of an Anesthetist

Several procedures performed in the electrophysiology laboratory (EP lab) require surgical manipulation and are lengthy. Patients undergoing such procedures usually receive general anesthesia or deep sedation administered by an anesthesiologist. In 536 consecutive procedures performed in the EP lab, we assessed the safety and efficacy of deep sedation administered under the direction of an electrophysiologist and in the absence of an anesthetist. Patients were monitored with pulse oximetry, noninvasive blood pressure recordings, and continuous ECGs. The level of consciousness and vital signs were evaluated at 5-minute intervals. Deep sedation was induced in 260 patients using midazolam, phenergan, and meperidine, then maintained with intermittent dosing of meperidine at the following mean doses: midazolam 0.031 ± 0.024 mg/kg; phenergan 0.314 ± 0.179 mg/kg; and meperidine 0.391 ± 0.167 mg/kg per hour. In the remaining 276 patients, deep sedation was induced with midazolam and fentanyl and maintained with a continuous infusion of fentanyl at a mean dose of 2.054 ± 1.43 μg/kg per hour. Fourteen patients experienced a transient reduction in oxygen saturation that was readily reversed following administration of naloxone. An additional 11 patients desaturated secondary to partial airway obstruction, which resolved after repositioning the head and neck. Fourteen patients experienced hypotension with fentanyl. All but one returned to baseline blood pressures following an infusion of normal saline. No patient required intubation and no death occurred. Only three patients had recollection of periprocedure events. No patient remembered experiencing pain with the procedure. Hospital stays were not prolonged as a result of the sedation used. In conclusion: (1) deep sedation during EP procedures can be administered safely under the guidance of the electrophysiologist without an anesthetist present; (2) the drugs used should be readily reversible in case of respiratory depression; and (3) this approach may reduce the overall cost of the procedures in the EP lab, maintaining adequate patient comfort.     

Radiofrequency Catheter Ablation of the Left and Right Ventricles: Anatomic and Electrophysiologic Observations

Certain untoward effects associated with the use of direct-current electrical catheter ablation of the ventricular endomyocardium have been noted. We assessed the efficacy and safety of closed-chest catheter ablation of the left and right ventricles using radiofrequency (RF) energy (750 kHz) in six dogs. Mean RF energies between 93 and 123 joules (J) were randomly delivered to three left ventricular (LV) sites via two distal adjacent electrodes (bipolar configuration) using 6-7F USCI tripolar or quadripolar catheters with an interelectrode distance of 5-10 mm. Another 90-143 J were given to two right ventricular (RV) sites in single or multiple divided applications between a distal electrode and an external patch electrode (unipolar configuration). Ventricular arrhythmias were not observed during application of RF energy. Programmed ventricular stimulation before and after the procedure did not induce ventricular tachycardia (VT) or fibrillation except in one dog who had inducible VT prior to ablation. There were no significant changes in LV and RV effective refractory periods after the procedures. Occasional premature ventricular beats and rare episodes of non-sustained VT (3-12 beats) were observed in ambulatory electrocardiographic recordings (13-24 hrs) done immediately after ablation. Dogs were sacrificed after 4-5 days. Pathology showed well-demarcated round or ovoid lesions of varying sizes. Mural thrombus was found in one dog. Microscopic findings consisted of circumscribed areas of coagulation necrosis with a peripheral zone of cellular infiltration. Transmural necrosis without perforation was occasionally seen in the thin RV wall when higher energies were delivered. In conclusion, discrete areas of desiccation injury in the ventricles can be achieved by transcatheter bipolar or unipolar ablation using RF energy. The complications associated with this method appear to be minimal. Further experiments are needed to evaluate its potential for catheter ablation of ventricular tachycardia.     

Eimeria tenella: quantitative in vitro and in vivo studies on the effects of mouse polyclonal and monoclonal antibodies on sporozoites

Murine, polyclonal and monoclonal antibodies, raised against sporozoites of Eimeria tenella, were tested for their ability to neutralize sporozoite infectivity in vitro and in vivo. Neutralization was effected via three mechanisms. Firstly, sporozoites fixed complement, at low titres, and lysis occurred by the alternative pathway of complement activation. Secondly, in the absence of complement activity, the murine heat-inactivated, hyperimmune antiserum neutralized sporozoites at relatively low titres. At high titres, even though sporozoites were agglutinated, neither the heat-inactivated hyperimmune antiserum nor the monoclonal antibody neutralized sporozoites. Finally, in the presence of complement and specific antibodies, at titres which by themselves would not neutralize sporozoites, neutralization was effected due to lysis via the classical pathway of complement activation.     

The Acridine Orange Viability Test Applied to Bone Marrow Cells I. Correlation with Trypan Blue and Eosin Dye Exclusion and Tissue Culture Transformation

Suspensions of murine bone marrow cells were stained with acridine orange(A-O) and observed under fluorescent microscopy after treatment withvarious injurious agents in order to establish the staining characteristics of"live" and "dead" cells. The percentage of viable cells demonstrated by the"A-O viability test" were correlated with eosin and trypan blue dye exclusionand tissue culture transformation viability tests. In general, the A-O testdemonstrated the viability of cells preserved by freezing as effectively as theother in vitro tests. In addition, the A-O test may be more sensitive indetermining the viability of cells where metabolic processes have been injuredby poisons or change in pH.

Submitted on September 4, 1963 Accepted on October 29, 1963     

Use of Fmoc-N-(2-hydroxy-4-methoxybenzyl) amino acids in peptide synthesis

The use of N, O-bisFmoc-N-(2-hydroxy-4-methoxybenzyl) amino acid derivatives in the synthesis of peptides with difficult sequences has already been described. With these amino acid derivatives the reversible protecting group 2-hydroxy-4-methoxybenzyl (Hmb) for the backbone amide bonds of peptide chains is introduced, and thus the aggregation due to hydrogen-bond interchain association is inhibited. This paper describes the synthesis and use of Fmoc-N-(2-hydroxy-4-methoxybenzyl)amino acid derivatives as an alternative means of introducing Hmb backbone protection. These new monoFmoc derivatives were obtained in higher yield than the bisFmoc derivatives. Coupling yields to the amino peptide resin were the same as those obtained with bisFmoc derivatives, under the TBTU/HOBt/DIEA conditions. We also compared different syntheses of a difficult peptide with the Fmoc approach [triple coupling, capping, use of chaotropic agents, backbone protection using monoFmoc (Hmb)Ala] and with optimized Boc chemistry. Both the backbone protection and optimized Boc chemistry approaches gave the desired product in excellent yield and purity. © Munksgaard 1997.     

Stability of subtilisins and related proteinases (subtilases)

The stability towards thermal and chemical (guanidine hydrochloride, GnHCl) denaturation of six inhibited subtilases (mesentericopeptidase, subtilisins BPN′, Carlsberg and DY, proteinase K and thermitase) has been investigated by kinetic and equilibrium studies. The unfolding processes were monitored by circular dichroic and fluorescence spectroscopy. Experiments in the absence and presence of extraneous calcium in the concentration range 2×10^?3-10^?1 M were performed. The presence of calcium in the weak calcium binding site changes the denaturation drastically. The heat- (or GnHCl-) induced unfolding curves obtained using CD spectroscopy show two independent transitions which seem not to have been resolved before. The presence of Ca²⁺ in the second (third in the case of thermitase) binding site increases the T_m, values by 11-21 °C and the δG_D(H₂O) values obtained from denaturation experiments in GnHCl by 6.7-7.2 kcal/mol when an extraneous Ca²⁺ concentration of 2 × 10^?2 M was used. One interpretation is that the initial step of denaturation in the presence of added calcium is the formation of a partially unfolded intermediate form, retaining a highly ordered structure with 60-85% of the a-helix structure of the native enzyme. This intermediate then unfolds at a temperature considerably higher than that of the same proteinases in the absence of added Ca²⁺. The free energy of stabilization of the intermediates is increased by 1.8-2.8 times in comparison with that for the unfolding reactions of the subtilases with empty Ca2/Ca3 binding sites. A second interpretation is that the two steps in the unfolding curves correspond to enzyme without and with calcium in the weak binding site. Fluorescence experiments confirm the mechanism involving the formation of intermediate states. The results are discussed in relation to the X-ray models of the six subtilases.