Potent and selective indolomorphinan antagonists of the kappa-opioid receptor.

The indole moiety in the delta-opioid antagonist, naltrindole (2, NTI), was employed as a scaffold to hold an "address" for interaction with the kappa-opioid receptor. The attachment of the address to the 5'-position of the indole moiety was based on superposition of NTI upon the kappa antagonist, norbinaltorphimine (1, norBNI). A variety of cationic groups were employed as a kappa address in an effort to investigate its interaction with the anionic address subsite, Glu297, on the kappa receptor. Some of the groups that were employed for this purpose were amines, amidines, guanidines, and quaternary ammonium. Members of the series were found to have a varying degree of kappa antagonist potency and kappa selectivity when tested in smooth muscle preparations. The 5'-guanidine derivative 12a (GNTI) was the most potent member of the series and had the highest kappa selectivity ratio. GNTI was 2 times more potent and 6-10-fold more selective than norBNI (1). In general, the order of potency in the series was: guanidines > amidines approximately quaternary ammonium > amines. The kappa antagonist potency appeared to be a function of a combination of the pK(a) and distance constraint of the cationic substituent of the ligand. Receptor binding studies were qualitatively in agreement with the pharmacological data. Molecular modeling studies on 12a suggested that the protonated N-17 and guanidinium groups of GNTI are associated with Asp138 (TM3) and Glu297 (TM6), respectively, while the phenolic hydroxyl may be involved in donor-acceptor interactions with the imidazole ring of His291. It was concluded that the basis for the high kappa selectivity of GNTI is related both to association with the nonconserved Glu297 residue and to unfavorable interactions with an equivalent position in mu- and delta-opioid receptors.

Influence of organic modifier concentration on plate number in micellar electrokinetic chromatography. 1. 2-propanol.

This paper establishes a physicochemical basis for the efficiency losses in micellar electrokinetic chromatography in buffers containing sodium dodecyl sulfate (SDS) and 2-propanol (2PN). Weakly, intermediately, and strongly retained analytes were separated in phosphate/borate buffers containing 50 mM SDS and from 0 to 10% 2PN by volume. Their plate numbers N generally agreed well with predictions of a theory for N based on longitudinal diffusion and instrumental contributions to dispersion. The N's of weakly and intermediately retained analytes were not affected strongly by 2PN over this concentration range, because their diffusion coefficients varied inversely with buffer viscosity and their retention times largely varied directly with viscosity. These combined effects on dispersion almost canceled. However, the N's of strongly retained analytes decreased with increasing 2PN, because their diffusion coefficients varied inversely with viscosity but their retention times increased more rapidly than did viscosity. These combined effects on dispersion did not cancel. These differences occurred because 2PN penetrated the micelles, caused bound counterions to be released, and increased the micellar charge and electrophoretic mobility. As 2PN concentration increased, the micelles electrophoresced increasingly rapidly against the electroosmotic flow. Consequently, strongly retained compounds required increasingly long times to elute.

Effect of a chemical modification on the hydrated adenosine intermediate produced by adenosine deaminase and a model reaction for a potential mechanism of action of 5-aminoimidazole ribonucleotide carboxylase.

Using the hydrated adenosine intermediate (6R)-6-amino-1, 6-dihydro-6-hydroxy-9-(beta-D-ribofuranosyl)purine (2) produced by adenosine deaminase (ADA, EC 3.5.4.4) as a starting point, the active site probe and inhibitor platform 5-(formylamino)imidazole riboside (FAIRs, 4) was designed by removal of the-C6(OH)(NH2)-molecular fragment of 2 generated by the early events of the enzyme-catalyzed hydrolysis. FAIRs was synthesized directly from the sodium salt of 5-amino-1-(beta-D-ribofuranosyl)imidazole-4-carboxylic acid (CAIR) along a reaction sequence involving a tandem N-formylation/decarboxylation that may have a mechanistic connection to the Escherichia coli purE-catalyzed constitutional isomerization of N5-CAIR to CAIR. The physical and spectral properties of FAIRs were elucidated, its X-ray crystal and NMR solution structures were determined, and its interaction with ADA was investigated. Crystalline FAIRs exists solely as the Z-formamide rotamer and exhibits many of the same intramolecular hydrogen bonding events known to contribute to the association of Ado to ADA. In water and various organic solvents, however, FAIRs exists as NMR-distinct, slowly interconverting Z and E rotamers. This truncated enzymatic tetrahedral intermediate analog was determined to be a competitive inhibitor of ADA with an apparent Ki binding constant of 40 microM, a value quite close to that (33 microM) of the natural substrate's K(m). The actual species selected for binding by ADA, though, is likely the minor hydroxyimino prototropic form of Z-FAIRs possessing a far lower true Ki value. As the structural features of FAIRs appear well-suited to support its use as a template for constructing active site probes of both ADA and AIR carboxylases, a variety of carbohydrate-protected versions of FAIRs suitable for facile aglycon elaborations were synthesized. The N3-alkylation, N3-borane complexation, and C4-iodination of some of these were investigated in order to assess physicochemical properties that may assist in the elucidation of mechanisms for the AIR carboxylases. The survey of these properties taken together with a reasonable mechanism for the model CAIRs-->FAIRs synthetic transformation is interpreted to support a mechanism for the purE-catalyzed N5-CAIR-->CAIR biosynthetic one that involves a carboxylative sp3-rehybridization of the imidazole C4 atom rather than one possessing a dipole-stabilized C4 sp2 carbanionic intermediate.

Relationship of 133Xe cerebral blood flow to middle cerebral arterial flow velocity in men at rest.

Cerebral blood flow (CBF) was measured by 133Xe clearance simultaneously with the velocity of blood flow through the left middle cerebral artery (MCA) over a wide range of arterial PCO2 in eight normal men. Average arterial PCO2, which was varied by giving 4% and 6% CO2 in O2 and by controlled hyperventilation on O2, ranged from 25.3 to 49.9 mm Hg. Corresponding average values of global CBF15 were 27.2 and 65.0 ml 100 g min-1, respectively, whereas MCA blood-flow velocity ranged from 42.8 to 94.2 cm/s. The relationship of CBF to MCA blood-flow velocity over the imposed range of arterial PCO2 was described analytically by a parabola with the equation: CBF = 22.8 - 0.17 x velocity + 0.006 x velocity2 The observed data indicate that MCA blood-flow velocity is a useful index of CBF response to change in arterial PCO2 during O2 breathing at rest. With respect to baseline values measured while breathing 100% O2 spontaneously, percent changes in velocity were significantly smaller than corresponding percent changes in CBF at increased levels of arterial PCO2 and larger than CBF changes at the lower arterial PCO2. These observed relative changes are consistent with MCA vasodilation at the site of measurement during exposure to progressive hypercapnia and also during extreme hyperventilation hypocapnia.

Human tolerance and physiological responses to exercise while breathing oxygen at 2.0 ATA.

Multiple physiological functions were monitored in ten men who performed two 30-min periods of 150-W ergometer exercise during 120-min exposures to O2 at 2.0 ATA. There were no convulsions or electroencephalographic manifestations of increased excitability. Sequential measurements of peripheral visual fields, pulmonary mechanical function, mental performance, and cardiovascular function during the resting recovery after each of the two exercise periods were not detectably altered from pre-exercise control values. Pre- and post-exposure measurements of visual acuity, accommodation, pupil diameter, visual cortical activity, and retinal electrical activity also revealed no significant differences. While CNS symptoms were absent, average arterial PCO2 rose by about 5 mm Hg during both exercise periods. This finding was confirmed in six subjects who performed four 6-min periods of continuous exercise at 50, 100, 150, and 200 W while breathing O2 at 2.0 ATA. Average arterial PCO2 rose nearly linearly from 34.3 mm Hg at rest to 44.0 mm Hg at 200 W. Arterial PCO2-related increments in brain blood flow and PO2 may explain part or all of the known detrimental influence of exercise on CNS O2 tolerance.

Correlative study of behavior and synaptic events during halothane anesthesia in the lamprey.

Behavioral anesthetic effects of halothane were compared with neurophysiological data obtained from monosynaptically coupled, single axon-giant neuron pairs in the isolated central nervous system of the lamprey in vitro. Based on the assumption that the number of halothane molecules at the site of anesthetic action is determined by molar concentration of the bath, depths of anesthesia were characterized in terms of synaptic events. The concentration producing anesthesia in 50% (AC50) and 95% (AC95) of the animals were 0.32 and 0.51 mM, respectively. With these concentrations, the depression of glutamate (GLUT)-mediated synaptic excitation remained less than 50%. Maximum suppression of the excitatory synaptic transmission was observed in the presence of 1.7 mM halothane which would be equivalent to the inhalation of 5.5% of this agent at 37 degrees C. At these concentrations, postsynaptic membranes did not respond to the bath-applied GLUT agonists. The decrease of agonist-induced responses to almost the same extent as excitatory postsynaptic potential (EPSP) amplitudes in the presence of halothane argues against a presynaptic mechanism affecting transmitter release. Persistent voltage-gated Na+ channel function at concentrations less than 2.4 mM is evidence against the involvement of this mechanism at these levels of anesthesia. The present results suggest that with depths between AC50 and AC95, anesthesia is associated with a partial reduction of sensitivity of the postsynaptic membrane to the excitatory amino acid transmitters.

Endothelium-dependent increase in vascular sensitivity to phenylephrine in long-term streptozotocin diabetic rat aorta.

1. The effect of short- and long-term streptozotocin (STZ)-induced diabetes (12 and 52 weeks) on the vascular response to phenylephrine was examined in the isolated thoracic aorta with and without intact endothelium from diabetic, age matched control rats and diabetic rats treated with insulin. 2. Twelve weeks after induction of diabetes, aortae with intact endothelium demonstrated no changes either in sensitivity (defined as pD2) or contractility (defined as the maximal developed tension per aortic tissue wet weight) to phenylephrine. 3. In contrast, 52 weeks after induction of diabetes, aortae with intact endothelium demonstrated an increased sensitivity to phenylephrine while contractility to phenylephrine was not changed. Insulin treatment partially corrected the increased sensitivity to phenylephrine observed in diabetic rat aorta. 4. Removal of endothelium abolished the difference in phenylephrine sensitivity between diabetic and control aortae at 52 weeks. 5. Pretreatment of intact aortae with methylene blue, an inhibitor of endothelium-derived relaxing factor (EDRF), abolished the difference in phenylephrine sensitivity between control and diabetic rat aortae at 52 weeks, while pretreatment with indomethacin, an inhibitor of cyclo-oxygenase, had no effect. These results suggest that decreases in production or release of EDRF might be responsible for the increased vascular sensitivity to phenylephrine observed in long-term STZ diabetic rats. 6. Acetylcholine-induced relaxation, which is EDRF-dependent, was less in diabetic rat aortae with intact endothelium at 52 weeks, but not at 12 weeks. These results further support the theory that decreases in capacity of the endothelium to synthesize or release EDRF may occur in long-term STZ diabetic rats.

Comparison of the anesthetic requirement for tolerance of laryngeal mask airway and endotracheal tube.

We tested the hypothesis that the laryngeal mask airway (LMA) is tolerated at lighter levels of anesthesia than an endotracheal tube (ET). We studied 20 unpremedicated, nonsmoking ASA physical status I or II patients aged 18-40 yr whose surgery lasted > 1 h. Subjects were randomly assigned to receive either an ET or LMA. Anesthesia was induced with intravenous propofol and the LMA or ET was inserted. The ET-group patients received 1.5 mg/kg of succinylcholine, preceded by vecuronium (0.015 mg/kg IV). Maintenance of anesthesia was with only isoflurane and approximately 66% N2O in O2 by spontaneous ventilation. All gas concentrations were measured by a Raman spectrometer sampling from the breathing circuit end of the LMA or ET. Toward the end of the procedure, the end-tidal N2O and isoflurane concentrations were allowed to decrease to < 3 vol% and 0.8 +/- 0.05 vol%, respectively. The end-tidal isoflurane concentration was then decreased in 0.1% +/- 0.05% decrements, each stable value being held for 5 min. The patient was observed for signs of reaction to the presence of the LMA or ET. The mean (range) end-tidal isoflurane concentrations for reaction to ET and LMA were 0.55% (0.4-0.7) and 0.35% (0.2-0.51), respectively (P < 0.001). These data confirm the original hypothesis of the study.

Controlled substance dispensing and accountability in United States anesthesiology residency programs.

Controlled substance dependence (CSD) among anesthesiology personnel, particularly residents, has become a matter of increasing concern. Opinions vary as to the effectiveness of controlled substances (CS) accountability in deterring, identifying, or confirming CSD. A survey of program directors of American anesthesiology training programs was conducted in the summer of 1990 to determine the level of CS dispensing and accountability within their programs. The survey demonstrated that CS dispensing and accountability varied considerably among programs, among hospitals associated with individual programs, and within geographically distinct anesthesia delivery areas within the separate hospitals. Nevertheless, most institutions were moving toward improved methods of CS dispensing and providing more and better CS accountability. The presence of significant CSD, particularly among anesthesiology residents, was reconfirmed. We were unable to correlate the level of accountability of CS with the incidence of CSD. It remains to be seen to what extent CS accountability will continue to develop and whether CSD prevalence will then be changed.

Surgical satellite pharmacies in institutions with anesthesiology training programs for physicians.

The results of a survey on the use of surgical satellite pharmacies in hospitals with anesthesiology training programs are reported. In June 1990 a questionnaire was mailed to 158 directors of anesthesiology training programs for physicians. The questionnaire solicited information on the presence of surgical satellite pharmacies in the training hospitals and the nature of the services provided, including accounting for controlled substances. Responses were received from 102 program directors and their designees, for a 65% response rate. Some respondents returned questionnaires completed by affiliated hospitals; a total of 137 responses were accumulated. Surgical satellite pharmacies were present in 46 (34%) of the 137 hospitals. Of those 46 satellite pharmacies, only 14 dispensed all controlled substances to anesthetic-administration areas. Most of the satellite pharmacies provided services at least eight hours per day. All 46 pharmacies dispensed controlled substances, 31 provided i.v. admixtures, and 12 dispensed all i.v. solutions. Accountability for controlled drugs was provided through a daily inventory count (45 satellite pharmacies), daily comparison of agents received and returned (36), review of the anesthesia record (31), random audits of individual providers (18), or quantitative or qualitative analysis of residual drugs (16). Accountability was considerably better in hospitals with surgical satellite pharmacies than in hospitals without them. Surgical satellite pharmacies provided increased accountability for controlled substances in institutions with anesthesiology training programs but did not use all the available methods for preventing drug diversion.

[Enhanced myocardial depression from bupivacaine in diabetic rats].

We examined the direct effects of bupivacaine on the diabetic rat myocardium using an isolated perfused (Langendorff) heart preparation. Rats were made diabetic by injection of streptozotocin (55 mg.kg-1, i. v.). 12 weeks after the administration of streptozotocin, hearts were excised and perfused using Langendorff technique. Following 60 min for equilibration, diabetic hearts from the 12-week animals exhibited significant bradycardia. Bupivacaine 3 mcg.ml-1 did not decrease myocardial contractility in control animals. However, 3 mcg.ml-1 bupivacaine decreased contractility significantly in diabetic animals at nearly all periods. Bupivacaine decreased heart rate and coronary flow significantly and similarly in control and diabetic rats hearts. After pacing at 4 Hz, bupivacaine decreased contractility more in diabetic group than in control group. These results indicate that the presence of a diabetic state appears to enhance the sensitivity of the heart to the myocardial depressant effect of bupivacaine.

Recovery from general anesthesia.

The pattern of recovery from anesthesia is ordinarily quite predictable. Using a limited number of drugs to provide anesthesia further improves predictability; it is more difficult to assign a cause for delayed emergence when multiple agents have been administered. We believe the recovery period is often more difficult to manage safely than induction and maintenance of anesthesia. During this phase, the effects of surgical trauma are superimposed on the patient's preexisting disease state, the anesthetic level is changing rapidly, respiratory function has not returned to normal, and circulatory reflexes may not have been regained. Because the patient is being moved from the operating room and care of the patient is being transferred from one person or team to others, vigilance and treatment may also be impaired. It is a period of care that warrants further study.

1H NMR study of the solution structure of the self-complementary dodecanucleotide d(TGCA)3.

The deoxyoligonucleotide d(TGCA)3 is a candidate for exhibiting unusual conformations. Its 1H NMR spectrum under low salt conditions has been obtained at 400 MHz and assigned using two-dimensional NMR techniques. The sugar puckers and glycosidic torsions have been determined by inspecting the relative intensities of the intranucleotide NOEs and COSY crosspeaks. At low electrolyte concentration (100 mM NaCl) the molecule exists as a right-handed duplex with twelve Watson-Crick base-pairs and deoxyribose moieties assuming the O1'-endo to C1'-exo pucker.

Isoflurane: an anesthetic for the eighties?

The introduction of isoflurane to clinical practice follows the search for a nonflammable, potent inhalation anesthetic which, above all, is chemically stable so as to resist biodegradation or attack by other chemicals. These attributes characterize isoflurane (Table 2). The hoped for freedom from hepatic and renal toxicity and from carcinogenic and mutagenic properties is a reality with this drug. Other favorable characteristics include relatively low solubility in blood in relation to anesthetic dose, lack of arrhythmogenic effect, provision of good muscle relaxation, and the absence of central nervous system excitation. Its moderate pungency detracts slightly from the ease of inhaled induction. Disadvantages include respiratory depression, reduced arterial blood pressure, uterine relaxation, decreased uteroplacental blood flow, and likely ability to trigger malignant hyperpyrexia. The frequency and/or significance of tachycardia and dilation of muscle blood vessels in clinical practice remain to be established. We believe isoflurane is a significant improvement over earlier potent inhalation anesthetics.