Hemicholinium-3: noncholinergic effects on squid axons.

Hemicholinium-3, when applied to the inside of a squid axon, is effective in blocking the action potential. This action is not antagonized by the addition of choline or acetylcholine to the perfusate. Voltage-clamp experiments show that hemicholinium-3 depresses both the early transient and late steady-state components of membrane ionic conductances, with a greater effect on the peak transient component.

Inhibition of potentially lethal radiation damage repair in normal and neoplastic human cells by 3-aminobenzamide: an inhibitor of poly(ADP-ribosylation).

The effect of 3-aminobenzamide (3AB), an inhibitor of poly(ADP-ribose) synthetase, on potentially lethal damage repair (PLDR) was investigated in normal human fibroblasts and four human tumor cell lines from tumors with varying degrees of radiocurability. The tumor lines selected were: Ewing's sarcoma, a bone tumor considered radiocurable and, human lung adenocarcinoma, osteosarcoma, and melanoma, three tumors considered nonradiocurable. PLDR was measured by comparing cell survival when cells were irradiated in a density-inhibited state and replated at appropriate cell numbers at specified times following irradiation to cell survival when cells were replated immediately following irradiation. 3AB was added to cultures 2 hr prior to irradiation and removed at the time of replating. Different test radiation doses were used for the various cell lines to obtain equivalent levels of cell survival. In the absence of inhibitor, PLDR was similar in all cell lines tested. In the presence of 8 mM 3AB, differential inhibition of PLDR was observed. PLDR was almost completely inhibited in Ewing's sarcoma cells and partially inhibited in normal fibroblast cells and osteosarcoma cells. No inhibition of PLDR was observed in the lung adenocarcinoma or melanoma cells. Except for the osteosarcoma cells, inhibition of PLDR by 3AB correlated well with radiocurability.

Cerebellar role in the load-compensating response of expiratory muscle.

The hypothesis that the cerebellum is involved in the load-compensating response of expiratory muscles to expiratory tracheal occlusion was tested in anesthetized cats. A continuous expiratory threshold load (ETL; 5 cmH2O) was applied to elicit consistent phasic baseline electromyographic activity in the transversus abdominis muscle (EMGab). Tracheal occlusion for single expirations (TOE) were applied, and the evoked responses were compared in the intact and decerebellate preparation. Cold blockade of the dorsal spinal column (C5-7) and bilateral vagal inactivation (cold blockade or transection) were employed to determine the role of afferents from the lung, airways, chest wall, and diaphragm in shaping the cerebellar involvement in the motor response. The results showed that 1) decerebellation increased the baseline amplitude of the integrated EMGab (fEMGab) activity (P < 0.05) with little change in expiratory duration, 2) TOE applied after decerebellation markedly increased the expiratory duration compared with the intact values (P < 0.05), with little effect on the peak fEMGab, 3) cooling the dorsal spinal columns (C5-7) did not significantly affect EMGab responses in the intact or decerebellate preparations, and 4) vagal inactivation in the intact or decerebellate preparation significantly eliminated the fEMGab responses to ETL and TOE. We conclude that the cerebellum is involved in the modulation of transversus abdominis activity during ETL and TOE. Vagal afferents provide the major sensory input for the cerebellar modulation of the expiratory loading response.

Role of phrenic nerve afferents in the control of breathing.

A long-held belief is that respiratory-related reflexes mediated by afferents in the diaphragm are weak or absent. However, recent data suggest that diaphragmatic afferents are capable of altering ventilatory motor drive as well as influencing perception of added inspiratory loads in humans. This review describes the sensory elements of the diaphragm, their central projections, and their functional significance in the control of respiratory muscle activation. The reflexes elicited by electrical stimulation of phrenic nerve afferents and the contribution of diaphragmatic afferents in respiratory load compensation and perception are considered. There is growing evidence that phrenic nerve afferents are activated under a variety of conditions. However, the significance of this input to the central nervous system is yet to be discerned.

A technique for recording from intact phrenic nerve afferents.

Recent evidence has suggested that phrenic nerve afferents can influence respiratory motor drive. This paper presents a technique whereby the activity of single phrenic nerve afferents can be recorded from uncut dorsal root filaments. Cervical dorsal roots 4, 5, and 6 were exposed by dorsal laminectomy in 10 anesthetized, spontaneously breathing cats. A stimulating electrode was placed on the right whole phrenic nerve low in the neck. The animal was then placed in a spinal suspension frame. Dissection of the dorsal root filaments was performed with probes made of fine tungsten wire. Single filaments were isolated intact from the dorsal root fascicles and placed across a tungsten electrode. Fiber classification was performed by determining conduction velocity. Monopolar recordings were made from a total of 38 fibers. Tonic activity was observed in 21, respiratory-related activity was evident in 15, and two fibers were silent but could be recruited by phrenic nerve stimulation. The conduction velocities ranged from 2.2 to 103 m/s. Approximately one-half of the fibers had conduction velocities of less than 20 m/s. This technique offers a way to record the activity of diaphragm afferents while maintaining the integrity of possible reflex pathways. Application of this method should prove helpful in elucidating the possible role of the various diaphragm afferents in the control of respiratory motor drive.

Diaphragm afferent modulation of phrenic motor drive.

Experiments were performed in eight lightly anesthetized thiopental sodium (Pentothal) cats to examine whether diaphragmatic afferents can significantly alter the neural drive to the diaphragm when the animal is exposed to lower body negative pressure. Moving-time-averaged diaphragmatic electromyograms (EMGma) were recorded and compared before and during exposure to lower body negative pressure in each of three consecutive conditions: C7 spinalization, bilateral vagotomy, and cervical dorsal rhizotomy. Application of lower body negative pressure in C7-spinalized animals resulted in a decrease in inspiratory time and peak diaphragmatic activity compared with control levels. After bilateral vagotomy, EMGma activity was prolonged with the application of lower body negative pressure. However, there was no increase in peak EMGma activity. After transection of the cervical dorsal roots subserving the phrenic nerve, the prolongation of diaphragmatic activity negative was eliminated. Therefore, we conclude that the significant increase in duration of inspiration in response to application of lower body negative pressure in the C7-spinalized, bilaterally vagotomized cat is mediated by phrenic nerve afferents.

Effect of nicotine and acetylcholine on crustacean muscle membrane potential.

We have investigated the effect of nicotine and acetylcholine on the resting membrane potential of the crayfish extensor muscle in order to determine whether crustacean muscle can be activated by cholinergic compounds. Intracellular recordings from individual deep extensor abdominal muscle cells were made using standard glass microelectrode techniques. The resting membrane potential was measured before and after treatment with glutamate, nicotine, and acetylcholine. Glutamate, which is a known activator of crayfish muscle, was used to determine whether the muscle cell preparation was viable and capable of responding to any of the test substances. Our results confirm that application of glutamate is associated with a depolarization of the muscle membrane. However, muscle cells showed no depolarization after treatment with nicotine (50 microM) or acetylcholine (66 microM). These results argue against the notion that increases in muscle tension may be responsible for the increased receptor organ discharge observed in the presence of nicotine. Rather, it supports the hypothesis that nicotine is acting directly on the mechanoreceptor membrane to change its sensitivity.

Involvement of the fastigial nuclei in vagally mediated respiratory responses.

Previous studies have demonstrated that the cerebellum, especially the fastigial nucleus (FN), is capable of modulating respiratory responses to chemical and mechanical stimuli. Because there is evidence to show projections from vagal afferents to the FN, the goal of this study was to determine the role of the FN in the respiratory reflexes elicited by activation of vagal afferents. Experiments were performed in anesthetized (chloralose), paralyzed, and artificially ventilated cats with an occipital exposure of the cerebellum. Administration of capsaicin (Cap; 5-10 micrograms/kg) via the right external jugular vein at the end of inspiration and application of lung inflation (LI; 10 cmH2O) during inspiration were carried out to stimulate nonmyelinated and myelinated vagal afferents, respectively. The phrenic neurogram was recorded as an index of the respiratory motor output. Control cardiorespiratory variables [expiratory duration (TE), arterial blood pressure] and their immediate responses to stimuli were compared before and after bilateral lesions of the FN. The results showed the following. 1) Cap injection and LI resulted in a dramatic increase in TE (apnea). 2) FN lesions did not significantly alter the control TE; however, the apneic duration induced by Cap injection was prolonged. 3) Neither FN lesions nor cerebellectomy affected the apneic duration that resulted from application of LI. 4) Cold blockade of the vagi (6-8 degrees C) eliminated the respiratory responses elicited by LI but not Cap injection; vagotomy abolished the responses to both stimuli. 5) FN lesions did not change the control ABP or its responses to either LI or Cap injection. It is concluded that the FN is involved in vagally mediated respiratory reflexes elicited by activation of nonmyelinated (C-fiber) vagal afferents.

Respiratory-related neurons of the fastigial nucleus in response to chemical and mechanical challenges.

Responses of cerebellar respiratory-related neurons (CRRNs) within the rostral fastigial nucleus and the phrenic neurogram to activation of respiratory mechano- and chemoreceptors were recorded in anesthetized, paralyzed, and ventilated cats. Respiratory challenges included the following: 1 ) cessation of the ventilator for a single breath at the end of inspiration (lung inflation) or at functional residual capacity, 2) cessation of the ventilator for multiple breaths, and 3) exposure to hypercapnia. Nineteen CRRNs having spontaneous activity during control conditions were characterized as either independent (basic, n = 14) or dependent (pump, n = 5) on the ventilator movement. Thirteen recruited CRRNs showed no respiratory-related activity until breathing was stressed. Burst durations of expiratory CRRNs were prolonged by sustained lung inflation but were inhibited when the volume was sustained at functional residual capacity; it was vice versa for inspiratory CRRNs. Multiple-breath cessation of the ventilator and hypercapnia significantly increased the firing rate and/or burst duration concomitant with changes noted in the phrenic neurogram. We conclude that CRRNs respond to respiratory inputs from CO2 chemo- and pulmonary mechanoreceptors in the absence of skeletal muscle contraction.

Modulation of respiratory motor output by cerebellar deep nuclei in the rat.

The present study was undertaken to determine what roles the various cerebellar deep nuclei (CDN) play in modulation of respiration, especially during chemical challenges. Experiments were carried out in 12 anesthetized, tracheotomized, paralyzed, and ventilated rats. The integrated phrenic nerve activity (integralPN) was recorded as an index of respiratory motor output. A stimulating electrode was sequentially placed into the fastigial nucleus (FN), the interposed nucleus, and the lateral nucleus. Only stimulation of the FN significantly altered respiration, primarily via increasing respiratory frequency associated with a pressor response. The evoked respiratory responses persisted after blocking the pressor response via pretreatment with phenoxybenzamine or use of transient stimulation (<2 s) but were abolished by microinjection of kainic acid into the FN. To test the involvement of FN neurons in respiratory chemoreflexes, ventilation with hypercapnic gases mixture and intravenous injection of sodium cyanide were applied before and after CDN lesions induced by kainic acid. CDN lesions did not significantly alter eupneic breathing, but FN lesions attenuated the respiratory response to hypercapnia and sodium cyanide. We conclude that, with respect to the CDN in the rat, FN neurons uniquely modulate respiration independent of cardiovascular effects and facilitate respiratory responses mediated by activation of CO(2) and O(2) receptors.

Chemosensitivity of crayfish slowly adapting stretch receptors to nicotine.

It has recently been demonstrated that slowly adapting stretch receptors (SASRs) in the airways of the dog respond directly to nicotine (Federation Proc. 43: 318, 1984). The purpose of the present experiment was to investigate this chemical effect on an isolated stretch receptor. The crayfish muscle receptor organ was chosen, since crayfish muscle is reported to be insensitive to nicotine or acetylcholine and therefore permits the testing of any direct chemical effect of nicotine on the muscle stretch receptors. The tail was removed and pinned out in a tissue bath, and a stretch receptor organ was surgically isolated. Single-unit SASR extracellular nerve recordings were made while simultaneously measuring tension in the tail. Drugs were prepared in Van Harreveld's solution and administered into the bath kept at 18 degrees C. When resting muscle tension was essentially reduced to zero by cutting both ends of the receptor organ muscle, nicotine (0.07 microM) added to the bath increased receptor activity fourfold. This response was abolished by treatment with hexamethonium (690 microM). In a second group of animals in which the muscle was left intact, nicotine was shown to significantly increase receptor sensitivity to step changes in muscle tension. Once again hexamethonium blocked the response to nicotine. These results demonstrate that the sensitivity of mechanoreceptor can be altered by chemical interaction with nicotinic receptors, which dramatically alter sensory receptor activity.

Cerebellar modulation of ventilatory response to progressive hypercapnia.

The cerebellar contribution to the ventilatory response to progressive hypercapnia was examined in 18 anesthetized tracheotomized spontaneously breathing cats. The absolute values for minute ventilation (VE), tidal volume (VT), respiratory frequency (f), inspiratory duty cycle (TI/TT), and mean inspiratory flow (VT/TI) were measured. Progressive hypercapnia [35-65 Torr end-tidal PCO2 (PETCO2)] was induced using the rebreathing method. The respiratory variables at each level of PETCO2 and the slopes of ventilatory (VT and f) responses to hypercapnia were compared across the intact, decerebellate, and decerebellate-vagotomized preparations. In 12 cats, decerebellation preceded vagotomy, and in 6 cats the order of the surgical procedures was reversed. The results show that, compared with intact control, decerebellation had little effect on respiratory variables when PETCO2 was 30-35 Torr. However, during a hypercapneic challenge (40-65 Torr PETCO2), VE and the slope of the VE response were significantly reduced. Bilateral vagotomy increased VT and decreased f but failed to alter the ventilatory response in the PETCO2 range of 35-55 Torr. However, combination of decerebellation and vagotomy, regardless of the surgical order, severely blunted VE (35-65 Torr PETCO2) and the slopes of VE, VT and f responses. When decerebellation followed vagotomy, significant decreases in VT (absolute values and slopes) were noted with little further alteration in f response. We conclude that the cerebellum and its interaction with the vagus nerves play a facilitatory or disinhibitory role in the ventilatory responses to hypercapnia.

Hypoxic respiratory responses attenuated by ablation of the cerebellum or fastigial nuclei.

The general contribution of the cerebellum to hypoxic respiratory responses and the special role of the fastigial nucleus (FN) in the hypoxic respiratory reflex mediated via peripheral chemoreceptors were investigated in anesthetized and spontaneously breathing cats. Seven cats were exposed to isocapnic progressive hypoxia before and after cerebellectomy by decreasing the fractional concentration of end-tidal O2 (FETO2) from 15 +/- 0.3% to 7% while maintaining the pressure of end-tidal CO2 at a constant level of approximately 30 mmHg. Five additional cats inhaled five breaths of pure N2 (transient hypoxia) and received sodium cyanide (50 micrograms iv) before and after thermal lesions of the bilateral FN. The results showed that cerebellectomy or FN lesions failed to alter the respiratory variables (minute ventilation, tidal volume, respiratory frequency, and the peak of integrated diaphragm activity) during eupneic breathing. However, cerebellectomy significantly attenuated minute ventilation (FETO2 < or = 13%) and the peak of integrated diaphragm activity (FETO2 < or = 10%) compared with control. During progressive hypoxia, changes in respiratory frequency were noted earlier (FETO2 < or = 13%) than changes in tidal volume (FETO2 < or = 10%). Similarly, bilateral lesions of the FN resulted in a profound reduction in these respiratory responses to transient hypoxia and sodium cyanide. We conclude that the cerebellum can facilitate the respiratory response to hypoxia and that the FN is an important region in the modulation of the hypoxic respiratory responses, presumably via its effects on inputs from peripheral chemoreceptors.

Transient respiratory augmentation elicited by acute head-down tilt in the anesthetized cat.

Acute head-down tilt (AHDT, -30 degrees) in humans induces a transient ventilatory augmentation for 1-2 min accompanied by a high venous return. However, the mechanisms underlying this respiratory response remain obscure because of limitations of experiments carried out in human subjects. The present study was undertaken to determine whether AHDT-induced respiratory augmentation exists in the anesthetized, paralyzed, and ventilated cat and, if so, whether this response depends on 1) the cerebellum, 2) the carotid sinus (CS) and/or vagal afferents, and 3) elevation of central venous return. The integrated phrenic neurogram, arterial blood pressure, central venous pressure (CVP), and end-tidal PCO2 were recorded before, during, and after AHDT. The results showed that AHDT produced a transient ( approximately 2 min) enhancement of minute phrenic activity (approximately 30%) primarily via an increase in peak integrated phrenic neurogram amplitude associated with a remarkable elevation of CVP (approximately 3 min). Cerebellectomy, CS denervation, bilateral vagotomy, or clamping CVP did not affect the presence of the AHDT-induced minute phrenic activity response. These findings demonstrate that the anesthetized cat is a suitable model for investigating the mechanisms involved in AHDT-induced respiratory augmentation. Preliminary studies suggest that this response does not require the cerebellum, CS/vagal afferents, or an associated rise in central venous return.

Microinjection of acetazolamide into the fastigial nucleus augments respiratory output in the rat.

The rostral fastigial nucleus (FNr) of the cerebellum facilitates the respiratory response to hypercapnia. We hypothesized that some FNr sites are chemosensitive to focal tissue acidosis and contribute, at least partially, to respiratory modulation. Minute ventilation (VE) was recorded in 21 anesthetized and spontaneously breathing rats. Acetazolamide (AZ; 50 microM) was microinjected unilaterally into the FNr while an isocapnic condition was maintained throughout the experiment. AZ (1 or 20 nl) injection into the FNr significantly elevated VE (46.0 +/- 6.7%; P < 0.05), primarily via an increase in tidal volume (31.7 +/- 3.8%; P < 0.05), with little effect on arterial blood pressure. This augmented ventilatory response was initiated at 6.3 +/- 0.8 min and reached the peak at 19.7 +/- 4.1 min after AZ administration. The same dose of AZ delivered into the interposed and lateral cerebellar nuclei, or vehicle injection into the FNr, failed to elicit detectable cardiorespiratory responses. To determine whether the ventilatory response to AZ injection into the FNr resulted from an increase in respiratory central drive, the minute phrenic nerve activity (MPN) was recorded in seven paralyzed and ventilated rats. Similar to VE, MPN was increased by 38.9 +/- 8.9% (P < 0.05) after AZ administration. Our results suggest that elevation of CO2/H+ within the FNr facilitates respiratory output, supporting the presence of ventilatory chemoreception in rat FNr.

Respiratory load compensation. II. Role of the cerebellum.

Effects of inspiratory tracheal occlusion (TO) on respiratory duration (inspiratory and expiratory duration), ventilation, and the peak integrated diaphragm electromyographic (integral of EMGdi) response were tested in 16 anesthetized cats before and after decerebellation with and without vagal input. The same protocols were repeated in the decerebrate preparation. Decerebellation did not significantly affect the baseline or the loaded values [tracheal occlusion (TO)] for respiratory duration, tidal volume, or magnitude of the integral of EMGdi response. Vagal blockade eliminated the load-compensating responses in the intact and the decerebrate preparation. However, vagal blockade in concert with decerebellation resulted in a significant (P < 0.05) reversible inhibition of the peak integral of EMGdi response during inspiratory TO. This suggests that removal of vagal and cerebellar influences during loaded breathing unmasked inhibitory inputs to the respiratory pattern generator. With vagus intact, decerebellation before or after decerebration abolished the attenuation of the peak integral of EMGdi response to TO observed with decerebration alone. We conclude that the cerebellum does play a role in determining the pattern of the respiratory response to TO. This influence may be direct and/or indirect via interaction with information emanating from suprapontine, vagal, and nonvagal sources.

Respiratory load compensation. III. Role of spinal cord afferents.

In a previous study, we reported that inspiratory tracheal occlusion (TO) significantly inhibited the motor drive to the diaphragm in a decerebellated bilaterally vagotomized preparation (J. Appl. Physiol. 75:675-681, 1993). The hypothesis to be tested in the present study was that respiratory muscle afferents activated by inspiratory TO provided the inputs responsible for the observed inhibition. Adult cats were anesthetized, tracheotomized, and instrumented with diaphragm electromyographic (EMGdi) recording electrodes. The cerebellum, vagi, and dorsal spinal cord (C2-T2) were surgically exposed. Inspiratory TO was applied before and after cold blockade of the dorsal cord (C6) or dorsal root (C3-6) transection in the intact and decerebellated vagotomized cat. Respiratory timing (inspiratory and expiratory duration) was determined from the EMGdi record, and the peak integrated EMGdi (integral of EMGdi) response was used as an index of respiratory motor drive. Our results showed that 1) cold blockade at the dorsal C6 level in an intact preparation significantly increased the peak of the integral of EMGdi response to TO and was reversible upon rewarming; 2) as previously reported, decerebellation coupled with bilateral vagotomy significantly decreased the peak integral of EMGdi response to TO with no effect on timing; 3) cold blockade (-1 degree C) of the dorsal cord at C6 significantly attenuated this inhibition, and subsequent dorsal rhizotomy at C3-6 completely abolished this inhibition; and 4) decerebellation, cold blockade of the dorsal cord (C6), and dorsal rhizotomy (C3-6) did not significantly affect baseline values in bilaterally vagotomized cats.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of diaphragm small-fiber afferent stimulation on ventilation in dogs.

Little is known regarding the role of diaphragm small-fiber afferents (groups III and IV) in the control of breathing. This study was designed to determine whether activation of these afferents with use of capsaicin affects phrenic efferent activity. Capsaicin injections into the phrenic artery were made in 10 alpha-chloralose-anesthetized dogs after each of the following procedures performed in succession: bilateral cervical vagotomy, C7 spinal cord transection, bilateral cervical dorsal rhizotomy. In six of these animals injections were also made after C2 spinal cord transection and removal of the cervical spinal cord. Injections made in the vagotomized animals were associated with apneusis followed by hyperpnea. C7 spinal transection eliminated the hyperpneic response, but the apneusis remained. Cervical dorsal rhizotomy or C2 spinal cord transection failed to abolish the apneusis in response to injection. No diaphragm response was obtained after removal of the cervical spinal cord. Experiments in three additional animals showed that capsaicin does not have a direct excitatory effect on the muscle cells of the crural diaphragm, nor does it potentiate the release of neurotransmitter in the diaphragm. The results of this study indicate that small-fiber afferents in the diaphragm have an excitatory effect on phrenic motoneurons. There is a segmental component to this reflex, since the response is observed after C2 spinal cord transection. The data also suggest that at least some of these afferents enter the spinal cord through the ventral roots.

Fastigial nucleus-mediated respiratory responses depend on the medullary gigantocellular nucleus.

Electrical stimulation of the rostral fastigial nucleus (FNr) alters respiration via activation of local neurons. We hypothesized that this FNr-mediated respiratory response was dependent on the integrity of the nucleus gigantocellularis of the medulla (NGC). Electrical stimulation of the FNr in 15 anesthetized and tracheotomized spontaneously breathing rats significantly altered ventilation by 35.2 +/- 11.0% (P < 0.01) with the major effect being excitatory (78%). This respiratory response did not significantly differ from control after lesions of the NGC via bilateral microinjection of kainic or ibotenic acid (4.5 +/- 1.9%; P > 0.05) but persisted in sham controls. Eight other rats, in which horseradish peroxidase (HRP) solution was previously microinjected into the left NGC, served as nonstimulation controls or were exposed to either 15-min repeated electrical stimulation of the right FNr or hypercapnia for 90 min. Histochemical and immunocytochemical data showed that the right FNr contained clustered HRP-labeled neurons, most of which were double labeled with c-Fos immunoreactivity in both electrically and CO(2)-stimulated rats. We conclude that the NGC receives monosynaptic FNr inputs and is required for fully expressing FNr-mediated respiratory responses.

Influence of nicotine in cigarette smoke on acute ventilatory responses in awake dogs.

To determine whether the acute ventilatory responses to inhaled cigarette smoke are affected by a difference in nicotine level, control cigarettes (low-nicotine research cigarettes) were laced with nicotine to generate an increase of 330% (mean) in nicotine content with little or no change in the levels of other smoke constituents. Acute ventilatory responses to both control and nicotine-laced cigarettes were determined and compared in six awake chronic dogs. Spontaneous inhalation of nicotine-laced cigarette smoke (10% concn, 750 ml vol) via a tracheostomy tube caused distinct and consistent changes in breathing pattern on the first or second breath of inhaled smoke: an apnea in three dogs, an augmented inspiration in two dogs, and rapid shallow breathing in one dog. No significant change in breathing pattern was found immediately following inhalation of control cigarette smoke. Both types of cigarettes caused a delayed hyperpnea. However, the increase in minute ventilation induced by nicotine-laced cigarettes (from a base line of 2.8 to a peak of 25.7 l/min) was significantly greater than that by control cigarettes (from 2.9 to 5.5 l/min). Results of this study suggest that nicotine is responsible for the elicitation of both the immediate and delayed ventilatory responses to inhaled cigarette smoke generated under our experimental conditions.