In Silico Prediction of Mechanism of Action for Cancer Therapeutics.

Cancer is currently the second leading cause of death in the U.S. and is projected to become the principal cause in the near future. While radiation and surgery are common cancer treatment methods, chemotherapy remains a key treatment option, offering distinct advantages over other therapy options, especially in the management of metastasized tumors. Understanding the mechanism of action (MoA) of current and newly developed drugs is crucial to ongoing drug development research. Foreknowledge of how a candidate drug works can yield a wealth of information, including which cancers a drug may treat more effectively based on the susceptibility of the cancer to drugs with the same MoA. Previous studies concerning prediction of MoA have relied on costly experimental measurements as input for their predictions. We have developed an a priori quantitative structure-activity relationship (QSAR) for the in silico prediction of MoA without the need for experimental measurements. This model enables us to relate structural features of a chemical to its efficacy with a predictive accuracy of over 80 %, thus identifying the MoA of a candidate drug without costly, time-consuming experimental tests.

Effect of different enhancers on the transdermal permeation of insulin analog.

Using chemical penetration enhancers (CPEs), transdermal drug delivery (TDD) offers an alternative route for insulin administration, wherein the CPEs reversibly reduce the barrier resistance of the skin. However, there is a lack of sufficient information concerning the effect of CPE chemical structure on insulin permeation. To address this limitation, we examined the effect of CPE functional groups on the permeation of insulin. A virtual design algorithm that incorporates quantitative structure-property relationship (QSPR) models for predicting the CPE properties was used to identify 43 potential CPEs. This set of CPEs was pre-screened using a resistance technique, and the 22 best CPEs were selected. Next, standard permeation experiments in Franz cells were performed to quantify insulin permeation. Our results indicate that specific functional groups are not directly responsible for enhanced insulin permeation. Rather, permeation enhancement is produced by molecules that exhibit positive logK(ow) values and possess at least one hydrogen donor or acceptor. Toluene was the only exception among the 22 potential CPEs considered. In addition, toxicity analyses of the 22 CPEs were performed. A total of eight CPEs were both highly enhancing (permeability coefficient at least four times the control value) and non-toxic, five of which are new discoveries.

A modified technique improved histology similarity to human intracranial aneurysm in rabbit aneurysm model.

Persistent undigested elastic membranes and collagen in the tunic media of rabbit elastase-induced aneurysm models lowered their histology similarity to human intracranial aneurysm. Our purpose was to make good the deficiency. Ten New Zealand white rabbits were divided into three groups: six rabbit in two groups for evaluating the difference in various treatments, and four rabbits for long-term observation. We inflated and occluded the right common carotid artery (CCA) by an endovascular technique. The first group of three rabbits was only given 200u elastase in the proximal segment of right CCA; the second group was given 100u elastase and 1.5mg collagenase, then the right CCA was ligated. The first and second groups were studied by magnetic resonance angiography (MRA), sacrificed at three weeks after aneurysm creation, their histology results were obtained and compared with human aneurysm. The third group underwent the same procedure as the second group only for three months of observation with enhanced MRA. Saccular aneurysms formed in all rabbits. Degeneration of the extracellular matrix and atrophy of smooth muscular cells in tunic media were more apparent in the second group. The third group remained stable for more three months. Two modifications included inflating the right CCA with a balloon and adding collagenase incubation can promote an aneurysm model more histologically similar to human aneurysm. In addition the improved aneurysm model remains stability for a long time.

A skin permeability model of insulin in the presence of chemical penetration enhancer.

Enhancing transdermal delivery of insulin using chemical penetration enhancers (CPEs) has several advantages over other non-traditional methods; however, lack of suitable predictive models, make experimentation the only alternative for discovering new CPEs. To address this limitation, a quantitative structure-property relationship (QSPR) model was developed, for predicting insulin permeation in the presence of CPEs. A virtual design algorithm that incorporates QSPR models for predicting CPE properties was used to identify 48 potential CPEs. Permeation experiments using Franz diffusion cells and resistance experiments were performed to quantify the effect of CPEs on insulin permeability and skin structure, respectively. Of the 48 CPEs, 35 were used for training and 13 were used for validation. In addition, 12 CPEs reported in literature were also included in the validation set. Differential evolution (DE) was coupled with artificial neural networks (ANNs) to develop the non-linear QSPR models. The six-descriptor model had a 16% absolute average deviation (%AAD) in the training set and 4 misclassifications in the validation set. Five of the six descriptors were found to be statistically significant after sensitivity analyses. The results suggest, molecules with low dipoles that are capable of forming intermolecular bonds with skin lipid bi-layers show promise as effective insulin-specific CPEs.

The effect of coculture on the postfertilization development of in vitro-matured monkey oocytes.

OBJECTIVE: To determine if the developmental potential of embryos resulting from in vivo- and in vitro-matured monkey oocytes could be increased through the use of a coculture system. DESIGN: Randomized prospective comparison of embryos resulting from either in vitro- or in vivo-matured oocytes cocultured with Vero cells or cultured in medium alone (control). SETTING: Basic research laboratory. MAIN OUTCOME MEASURES: In vitro embryo development to the blastocyst stage and blastocyst hatching. RESULTS: No significant difference in development was noted between coculture and control groups with embryos resulting from in vivo-matured oocytes. However, coculture was found to improve significantly the development of monkey embryos resulting from in vitro-matured oocytes. CONCLUSIONS: These results demonstrate that primate embryos resulting from in vitro-matured and in vitro-fertilized oocytes differ in their culture requirement when compared with embryos resulting from in vivo-matured oocytes.

One-way neonatal transports: a new approach to increase effective utilization of air medical resources.

INTRODUCTION: High-risk neonates often require significant stabilization and preparation time for transport. The purpose of this study was to determine whether the institution of a formal neonatal transport policy would increase the effective utilization of air medical resources and to determine whether such a policy would be useful to other air medical transport programs. METHODS: A descriptive review of flight data from time periods before and after the institution of the policy, combined with a telephone survey of 20 other rotor-wing transport programs. RESULTS: After implementation of the policy, the total number of neonatal transports decreased (85 in 13 months vs 60 in 17 months) as did the number of two-way transports per month (4.6 vs 1.3). In addition, average mission time for all neonatal transports decreased. Fifteen of 20 transport programs in this Association of Air Medical Resources region perform neonatal transports. Only one of the 15 has a formal policy to determine the use of one-way versus two-way neonatal transports. CONCLUSION: The neonatal team transport policy has increased the effective utilization of air medical resources in this program. This policy could have wider application for other programs as well.

Quantitative effects of sympathetic and vagal nerve stimulations on sinus and AV junctional rhythms.

Responses of the sinus node and atrioventricular (AV) junctional pacemakers to autonomic denervation and to individual stimulations of the right and left stellate and both vagi were studied in 33 anesthetized dogs. Autonomic denervation depressed sinus node automaticity by only 18% from control, whilst AV junctional automaticity was reduced by 48.5% from control. Sympathetic and parasympathetic stimulation frequency-response curves (0.25, 0.5, 1, 2, 4, 8, 16 and 32 Hz) were obtained. In the sinus node the chronotropic responses to sympathetic stimulations reflect a bilaterally asymmetrical innervation with a right sided preponderance. In contrast, sinus slowing in response to either right or left vagal stimulations were indistinguishable when lower frequencies of stimulation were used. At 4 Hz and higher frequencies there is a right vagal preponderance. The AV junctional chronotropic responses suggest that this major subsidiary pacemaker receives a bilaterally symmetrical autonomic innervation. The chronotropic responses to individual nerve stimulations expressed as percent changes in sinus rate and AV junctional rate from their respective controls after autonomic denervation show that the AV junction is far more responsive than the sinus node to both sympathetic and parasympathetic stimulations. To allow for more meaningful comparisons the data were normalized using the respective maximum increase and maximum decrease of sinus node and AV junctional rates to left and right sympathetic and parasympathetic stimulations as the 100% reference. These normalized curves show that 50% of the maximal chronotropic responses were always achieved at a lower stimulus frequency in the AV junction than in the sinus node; shift of the AV junctional response curves to the left of the sinus node response curves by a 0.2 (sympathetic) and 0.3 (parasympathetic) log units was observed. These studies further showed that sympathetic activity in the AV junction is an absolute prerequisite to maintain regular AV junctional rhythms especially during the bradycardic episodes evoked in the study of vagal stimulus frequency-response curves.

Differential cardiac sympathetic activity during acute myocardial ischemia.

Efferent sympathetic activities were simultaneously recorded from two thoracic cardiac nerves in 33 chloralose-anesthetized dogs. Efferent innervation patterns were determined by electrical stimulation prior to recording in each animal. One of the nerves selected for recording was shown to innervate the proposed ischemic region, whereas the other nerve was selected because it was shown to innervate nonischemic regions. Left ventricular ischemia was produced by occlusion of a branch of either the left anterior descending (LAD) or left circumflex (LCX) coronary arteries. Heart rate was paced. Cardiac postganglionic sympathetic efferent activities were recorded during a 30-min coronary occlusion in 22 animals. Thirty minutes after LAD occlusion (n = 10), postganglionic sympathetic activity to ischemic myocardium was decreased (84 +/- 5% of control; P less than 0.05) while activity to nonischemic myocardium was unchanged. Thirty minutes after LCX occlusion (n = 12), postganglionic sympathetic activity to ischemic myocardium was also decreased (87 +/- 3% of control; P less than 0.01); however, sympathetic activity to nonischemic myocardium was increased (159 +/- 10% of control; P less than 0.001). Thus, in the anesthetized canine, regional left ventricular ischemia elicits differential sympathetic neural responses that are dependent on the location of the ischemic myocardium as well as the efferent destinations of the nerves. Changes in cardiac postganglionic sympathetic efferent activities are characterized by decreased activity to ischemic regions, with either no change or increased activity to nonischemic regions.

Effects of deafferentation or sequential occlusions on cardiac sympathetic activity during ischemia.

In 39 anesthetized dogs, we compared the effects of selective afferent sympathectomy, vagotomy, epicardial phenol painting, or a previous coronary occlusion on cardiac sympathetic activities recorded during 30 min of acute myocardial ischemia. Efferent innervation patterns were verified by electrical stimulation, and cardiac efferent activities were simultaneously recorded from two thoracic sympathetic nerves. One nerve was selected for recording because it was shown to innervate the proposed ischemic region, whereas the other nerve was selected because it was shown to innervate nonischemic regions. Left ventricular ischemia was produced by occlusion of a small branch of either the left anterior descending (LAD) or left circumflex (LCX) coronary arteries. Heart rate was paced. Reflex changes in sympathetic activities to ischemic regions were prevented by elimination of afferent fibers with either phenol or bilateral vagotomy. Ablation of sympathetic afferents prevented ischemia-induced reflex changes to nonischemic regions but did not prevent reflex changes to ischemic regions. Reflex changes in cardiac sympathetic activities during a second coronary occlusion of 30 min were vastly different from the initial occlusion but were similar to those observed after local deafferentation. Our findings indicate that 1) differential reflex changes in cardiac sympathetic activities during 30 min of left ventricular ischemia are due to afferent signals originating from the ischemic region, 2) both sympathetic and vagal afferent fibers participate in the ischemia-induced cardio-cardiac reflex, and 3) the nature of the sympathetic reflex response to acute ischemia is influenced by a previous ischemic insult.

The effects of negative chronotropic interventions on sinus node recovery time.

The effects of multiple increases in sinus cycle length on sinus node recovery time (SNRT) were examined in 5 dogs. Pacing was performed from the left atrial appendage for 30 and 60 seconds using at least 4 different pacing cycle lengths selected between 230 and 620 msec. Each dog received propranolol (1 mg/kg, IV) prior to any measurements. The effects of increases in sinus cycle length on SNRT were first assessed during 2 levels (4 and 8 Hz) of continuous vagal stimulation. From a control cycle length of 439 +/- 28 msec (mean +/- SE), the vagal stimulations lengthened the sinus cycle lengths to 604 +/- 10 msec and 758 +/- 16 msec respectively. Sinus cycle length was then prolonged by combined muscarinic and beta-receptor blockade resulting in a sinus cycle length of 549 +/- 9 msec. Autonomic blockade plus verapamil (3-10 mg IV) resulted in sinus cycle lengths of 612 +/- 14 and 721 +/- 18 msec respectively, which were not significantly different from those obtained with vagal stimulation. Data relating SNRT to the sinus cycle length, pacing cycle length, duration of pacing and the negative chronotropic interventions used to achieve the changes in the sinus cycle length were analyzed via covariance analysis. The results demonstrate that the single most important determinant of SNRT is the sinus cycle length. Furthermore, equivalent increases in sinus cycle length whether obtained by vagal stimulation, autonomic blockade or intravenous verapamil results in SNRTs that are not significantly different. Therefore, in the sinus node, changes in the rate of pacemaker activity, regardless of how they are achieved, will largely determine the changes in SNRT.

Differential interaction of adrenergic and cholinergic effects on AV junctional automaticity and AV conduction.

The effects of postsynaptic autonomic interactions on atrioventricular (AV) junctional automaticity and AV conduction were studied in six canine heart in situ using direct injections of norepinephrine (NE) and physostigmine (PSM) into the AV node artery. Injection of NE (0.05 microgram/ml, 2 ml) caused an AV junctional rhythm (AVJR) in every dog. After injection of PSM (10 micrograms/ml, 2 ml), the responses of AVJR to NE were virtually identical to those observed before cholinesterase inhibition (160 +/- 13 vs 162 +/- 12 bpm). In contrast, this moderate cholinesterase inhibition still had a readily demonstrable negative dromotropic effect. In any given dog, depressed AV conduction was characterized by one of two types (I and II) of retrograde atrial capture during AVJR. Before PSM in the AV junction, onset of atrial depolarization during AVJR preceded the onset of ventricular depolarization in both type I and type II responses. After PSM, atrial depolarization occurred later with respect to ventricular depolarization (i.e., during or mostly after ventricular activation) in type I, whereas in the type II responses atrial depolarizations began much earlier than before PSM, thus being completed long before the onset of ventricular activation. Because of such differential responsiveness of AV junctional automaticity and AV conduction and because of the two types of intranodal conduction observed after administration of PSM into the AV junction, we can postulate that under appropriate autonomic imbalance retrograde or antegrade AV block could readily develop in spite of preserved AV junctional automaticity.

Atrioventricular junctional tachycardia during heart block.

The response of the atrioventricular (AV) junction to brief intense adrenergic stimulation applied during episodes of second degree heart block achieved by acetylcholinesterase paralysis in the AV junction was examined in six dogs. Despite profound depression of AV conduction due to enhanced cholinergic activity, strong local adrenergic stimulation still readily elicited AV junctional tachycardia. Increase in cholinomimetic influences in the AV junction did not prolong transatrial or His bundle-ventricular conduction times. During AV junctional rhythm and retrograde atrial capture (n = 4), neither the sequence of retrograde atrial activation nor the atrial electrogram configurations were altered. In the two remaining dogs the AV junctional tachycardia was associated with AV dissociation. These findings suggest that the acetylcholine-induced depression of AV conduction is located in the AV node region exclusively. More important, however, is the demonstration that retrograde atrial activation originating from a pacemaker located in the AV node or immediate vicinity could actually precede the inscription of the H spike by a considerable amount of time, further suggesting that anterograde conduction from the pacemaker site to the bundle of His is far more depressed by acetylcholine than is the concomitant retrograde conduction from the pacemaker site to the atrium. Thus, inference of the origin of a subsidiary pacemaker from the P wave configuration or the relation of the A wave to the His bundle electrogram, or both, may lead to erroneous conclusions.

Differential effects of sympathetic activity on AV junctional automaticity and AV conduction.

Rapid ventricular response during episodes of supraventricular tachycardia are often followed, on abrupt cessation of the tachycardia, by prolonged pauses terminated by a sluggish and sometimes erratic escape of a supraventricular pacemaker. Such chronotropic-dromotropic paradoxes are readily reproduced in the animal laboratory following elimination of the sinus node and bilateral decentralization of the stellate ganglia and vagi. This study examined whether left stellate stimulation (0.5, 1, 2, 4, 8 and 16 Hz) or lack thereof differentially affected AV junctional automaticity and AV conduction. In the absence of any sympathetic neural activity (maximal sympathetic deficit), the AV junctional rate averaged a mere 22 +/- 2 percent of its peak performance, whereas under the same conditions, anterograde AV conduction averaged 73 +/- 5 percent and retrograde VA conduction 56 +/- 13 percent of their respective peak performances. On comparing the response curve (normalized responses) for AV junctional automaticity with that obtained for anterograde AV conduction the differences were significant at all frequencies between 0 and 4 Hz. Retrograde VA conduction (as assessed by the fastest ventricular pacing rate still conducted 1:1 to the atria) was always significantly less than anterograde AV conduction (as assessed by the fastest atrial pacing still conducted 1:1 to the ventricles). These results indicate that AV junctional automaticity is considerably more affected by sympathetic deficit than are either anterograde or retrograde AV conduction. In other words, AV junctional automaticity is far more dependent upon sympathetic input than AV conduction. While sympathetic influence is critical to the escape and maintenance of AV junctional automaticity both anterograde and retrograde AV conduction are remarkably resilient even under conditions of severe sympathetic deficit.

Cardiac autonomic efferent activity during baroreflex in puppies and adult dogs.

Blood pressure and heart rate were recorded in 15 anesthetized puppies (6-10 wk, 1-6 kg) and 18 adult mongrel dogs (greater than 1 yr, 18-26 kg) before and during acute blood pressure changes achieved with nitroglycerin or phenylephrine (4 and 8 micrograms/kg iv). Overall heart rate responses to blood pressure changes in adults were significantly (P less than 0.05) greater than those in puppies. Following control baroreflex responses, two multifiber efferent preparations from the discrete thoracic cardiac nerves (sympathetic, n = 48; parasympathetic, n = 18) were simultaneously recorded and analyzed by microprocessor. Severing of the nerves significantly attenuated the heart rate responses to blood pressure changes in puppies only, suggesting less redundancy of the neural regulation of the sinus node in the puppy. The pressure-induced reflex changes in the sympathetic or parasympathetic efferent nerve activities were not significantly different between adult dogs and puppies. There were no significant differences in reflex activities in right-sided (n = 29) vs. left-sided (n = 19) sympathetic nerves in either puppies or adult dogs. Preganglionic sympathetic fibers in puppies (but not adult dogs) were more responsive to blood pressure changes than were postganglionic sympathetic fibers. Thus baroreceptor reflex control in the puppy is less developed than in the adult canine heart, and the maturational difference in neural regulation of the heart is at or beyond the efferent nerve terminals.

Differential sympathetic-parasympathetic interactions in sinus node and AV junction.

This study examined the effects and interactions evoked by various combinations of parasympathetic (P) and sympathetic (S) stimulations on sinus node automaticity and atrioventricular (AV) conduction (both anterograde and retrograde) in 11 two-mo old anesthetized puppies. The respective effects of these autonomic maneuvers were assessed by covariant analysis using a linear regression model of the form Y = alpha 0 + alpha 1 X base line + alpha 2 X sequence + alpha 3 X S + alpha 4 X S2 + alpha 5 X P + alpha 6 X P2 + alpha 7 X (P X S).... The effects of parasympathetic and sympathetic stimulation, of the quadratic term S2, and of the interaction term (P X S) were highly significant (P less than 0.0001) on both sinus node automaticity and AV conduction (anterograde and retrograde). In contrast, the effect of the quadratic term P2 was significant on sinus node automaticity only. Sequence of stimulation whether (SP) or (PS) had no significant effect on either chronotropic or dromotropic properties. Furthermore, whereas augmented antagonism with parasympathetic predominance over sympathetic activity was readily demonstrable in the control of sinus rate, the reverse, i.e., augmented antagonism with sympathetic preponderance over parasympathetic activity, was found to apply to the control of AV conduction (both anterograde and retrograde). Hence, the nature and extent of parasympathetic-sympathetic interactions vary considerably, depending on which cardiac structure and/or function is under scrutiny. It appears that AV conduction is especially sensitive to modulation of sympathoadrenergic activity, whereas sinus node automaticity is particularly responsive to cholinomimetic influences.

Effects of the diethylamino analog of ethmozin (ethacizin) upon sympathetic and parasympathetic efferent activity to the canine heart.

This study examined the effects of the diethylamino analog of ethmozin (ethacizin) (1 mg/kg i.v.) on the spontaneous and reflexly elicited efferent activity in thoracic cardiac sympathetic and parasympathetic nerves. Nitroglycerin and phenylephrine (4 and 8 micrograms/kg i.v.) were administered to 15 anesthetized mongrel dogs while monitoring blood pressure and heart rate. In each dog two cardiac nerves were isolated and efferent neurograms were recorded simultaneously and analyzed by microprocessor. Ethacizin significantly attenuated the spontaneous sympathetic efferent activity in both left and right, preganglionic (n = 8) and postganglionic (n = 14) sympathetic nerves to the heart. In contrast, reflex changes in sympathetic activity elicited by baroreceptor challenges were not affected by ethacizin. Also, ethacizin did not significantly affect either spontaneous or baroreceptor reflex-induced parasympathetic efferent activities in eight preganglionic nerves. Thus, this new phenothiazine derivative may exert part of its antiarrhythmic action through a reduction of the spontaneous sympathetic tonic discharges to the heart. The fact that ethacizin reduced neither the reflex-induced changes in sympathetic or parasympathetic activities nor influenced the tonic vagal discharges further suggests that the compound is not likely to interfere with reflexly mediated cardiovascular adaptive changes.

Differences in the determinants of overdrive suppression between sinus rhythm and slow atrioventricular junctional rhythm.

Sinus node recovery time was compared to the recovery time of a slow atrioventricular junctional rhythm in each of the same seven pentobarbital anesthetized dogs. Recovery time and the first five cardiac cycles were examined after pacing atria and ventricles for 20, 40, and 60 seconds at four or more pacing cycle lengths. Data relating recovery times and return to control conditions to prepacing cycle length, pacing cycle length, duration of pacing, site of pacing, and origin of rhythms were analyzed by covariance analysis. From the analyses, the relative contribution of the determinants are: the prepacing cycle length 73%, the site of pacing 3.5%, the pacing cycle length 2%, and the interaction of the site of pacing and pacing cycle length 1% for sinus node recovery time; and for slow atrioventricular junctional rhythm recovery time, the duration of pacing 40%, the interactions between the duration of pacing and the pacing cycle length 27%, and the prepacing cycle length 9%. A modified exponential decay model predicted 8 beats for return to prepacing conditions during sinus rhythm and 66-100 beats during atrioventricular junctional rhythm. We conclude that the single most important determinant of sinus node recovery time is the prepacing cycle length. Pacing cycle length and site of pacing have a significant but small influence on sinus node recovery time and duration of pacing, beyond 20 seconds, has no significant influence. In contrast, duration of pacing is the most important determinant of slow atrioventricular junctional recovery time. Another major determinant of slow atrioventricular junctional recovery time is the interactions between pacing cycle length and duration of pacing. Prepacing cycle length has a minor influence, and site of pacing has no influence, on slow atrioventricular junctional recovery time.

Negative chronotropic and parasympatholytic effects of alinidine on canine sinus node and AV junction.

The direct effects of alinidine (N-allyl-clonidine) on the sinus node and atrioventricular (AV) junction were studied in 18 anesthetized dogs. Stimulus frequency-response curves to right stellate ganglion and right cervical vagus stimulations as well as responses to norepinephrine or acetylcholine were determined before and after selective perfusion of alinidine into the sinus node artery. Alinidine (1 microgram/ml) had no effect on spontaneous sinus rate [148 +/- 5 (SE) beats/min]. However, alinidine concentrations of 5, 10, and 25 micrograms/ml produced significant (P less than 0.05) sinus slowing to 138, 127, and 121 beats/min, respectively. Recovery to control rate was dose dependent and took from 4 to 33 min. Sinus rate increases with right stellate stimulations were not affected by alinidine. However, sinus rate decreases with right vagal stimulations were significantly (P less than 0.01) attenuated by alinidine. The negative chronotropic effects of acetylcholine were not influenced by alinidine. Alinidine (1-100 micrograms/ml into AV node artery) had no effect on the A-H interval of the His bundle electrogram. However, alinidine (10 and 25 micrograms/ml) diminished the AV block produced by stimulation of the left vagus in electrically paced hearts but not the negative dromotropic actions of directly administered acetylcholine. Thus alinidine has direct negative chronotropic effects, no effect on sinus node responses to sympathetic stimulation, ability to diminish sinus node and AV junctional responses to vagal stimulations without interference at the cholinergic muscarinic receptor, and 4) no effect on AV nodal conduction.

Influence of atrial mechanoreceptors upon the sympathetic efferent activity elicited during a cardiogenic chemoreflex in the dog.

Two different cardiogenic reflexes were studied for their effects upon the activity recorded from the thoracic cardiac nerves of 7 chloralose-anesthetized dogs. Sympathetic efferent activity in 5 right and 7 left anterior ansa multifiber preparations was quantitatively analyzed. A cardiogenic hypertensive chemoreflex was induced by administration of serotonin (100 micrograms/ml, 2 ml) into the left atrium before, during and after inflation of balloons in the right and/or left atrial appendages. Balloon inflations alone resulted in heart rate increases of 5-20 bpm but arterial blood pressure, pulse pressure and end diastolic pressure were not significantly affected. Control serotonin-induced efferent discharges were 174 +/- 10% (mean +/- S.E.M.) of resting activity with a duration of 5.2 +/- 0.4 s. During inflation of both atrial balloons, the serotonin-induced efferent discharges were 165 +/- 14% (N.S.) of resting activity with a duration of 7.8 +/- 0.7 s (P less than 0.01). Following release of the balloons, the serotonin-induced discharges were 152 +/- 17% (N.S.) of resting activity with a duration of 4.9 +/- 0.4 s (N.S.). These results suggest that afferent signals from atrial mechanoreceptors can act to increase the duration but not the total intensity of the efferent sympathetic discharges elicited during a cardiogenic chemoreflex.

Effects of gamma-aminobutyric acid on neural regulation of the canine sinus node.

The direct effects of gamma-aminobutyric acid (GABA) on the sinus node and its neural regulation were studied by selective perfusion of the sinus node artery (SNA) in 26 open-chest pentobarbital-anesthetized dogs. GABA (1-1,000 micrograms/ml, 2 ml, SNA) produced no direct effect on intrinsic sinus rate, but in several experiments (at 1 microgram/ml) it produced an inconsistent augmentation of the sinus tachycardia due to right stellate stimulation. At all other concentrations tested (10-1,000 micrograms/ml), GABA did not significantly alter the sinus node response to stellate stimulation. Local alpha-receptor blockade with phentolamine or muscarinic receptor blockade with atropine through the SNA prior to the perfusion of GABA did not change the sinus node responses. GABA (10-1,000 micrograms/ml, 2 ml, SNA) significantly attenuated (by 45-80% of control) the sinus bradycardia due to stimulation of the right vagosympathetic trunk. The sinus node response to selective perfusion of norepinephrine or acetylcholine (0.01 micrograms/ml, 2 ml, SNA) was not affected by GABA. The stellate augmentation (when present) and vagal attenuation were both antagonized by perfusion of picrotoxin (1,000 micrograms/ml, 2 ml, SNA). It is concluded that GABA does not exert any direct effect on the sinus node but may indirectly influence sinus rate by an effect on local nerves or ganglia.