Early exposure to farm dust in an allergic airway inflammation rabbit model: Does it affect bronchial and cough hyperresponsiveness?

INTRODUCTION: Over the past 50 years, the prevalence of allergic respiratory diseases has been increasing. The Hygiene hypothesis explains this progression by the decrease in the bio-diversity of early microbial exposure. This study aims to evaluate the effect of early-life farm exposure on airway hyperresponsiveness and cough hypersensitivity in an allergic airway inflammation rabbit model. METHOD: A specific environment was applied to pregnant rabbits and their offspring until six weeks after birth. Rabbits were housed in a pathogen-free zone for the control group and a calf barn for the farm group. At the end of the specific environmental exposure, both groups were then housed in a conventional zone and then sensitized to ovalbumin. Ten days after sensitization, the rabbit pups received ovalbumin aerosols to provoke airway inflammation. Sensitization to ovalbumin was assessed by specific IgE assay. Cough sensitivity was assessed by mechanical stimulation of the trachea, and bronchial reactivity was assessed by methacholine challenge. The farm environment was characterized by endotoxin measurement. RESULTS: A total of 38 rabbit pups were included (18 in the farm group). Endotoxin levels in the farm environment varied from 30 to 1854 EU.m-3. There was no significant difference in specific IgE values to ovalbumin (p = 0.826) between the two groups. The mechanical threshold to elicit a cough did not differ between the two groups (p = 0.492). There was no difference in the number of cough (p = 0.270) or the intensity of ventilatory responses (p = 0.735). After adjusting for age and weight, there was no difference in respiratory resistance before and after methacholine challenge. CONCLUSION: Early exposure to the calf barn did not affect cough sensitivity or bronchial reactivity in ovalbumin-sensitized rabbits. These results suggest that not all farm environments protect against asthma and atopy. Continuous exposure to several sources of microbial diversity is probably needed.

A Physical Analog to Assess Surgical Face Mask Air Flow Resistance During Tidal Ventilation.

A large variety of disposable face masks have been produced since the onset of the COVID-19 pandemic. Decreased resistance to inspiration improves adherence to the use of the mask; the so called breathability is usually estimated by the measurement of air flow across a section of the tissue under a given pressure difference. We hypothesized that the mask pressure-flow relationship studied in conditions that mimic tidal breathing could allow a more comprehensive characterization of airflow resistance, a major determinant of mask comfort. A physical analog was made of a plaster cast dummy head connected through a pneumotachograph to a series of bellows inflated/deflated by a respirator. Pressure was measured at the mock airway opening over which the mask was carefully secured. The precision of the measurement equipment was quantified using two estimates of measurement error: repeatability coefficient (RC) and within-mask coefficient of variation (CVwm). The airflow resistance of 10 surgical masks was tested on 4 different days. Resistance means did not differ significantly among four repeated measures (0.34 hPa.s.L-1; 0.37 hPa.s.L-1; 0.37 hPa.s.L-1; and 0.37 hPa.s.L-1; p = 0.08), the estimated RC was 0.08 hPa.s.L-1 [95%CI: 0.06-0.10 hPa.s.L-1], and CVwm was 8.7% [95%CI: 1.5-12.2%]. Multiple comparisons suggest the presence of a learning effect by which the operator reduced the error over the course of repetitive resistance measurements. Measurement precision improved considerably when the first set of measures was not taken into account [RC ~ 0.05 hPa.s.L-1 (95%CI: 0.03-0.06 hPa.s.L-1); CVwm~4.5% (95%CI: 1.9-6.1%)]. The testing of the face mask resistance (R) appears simple and highly repeatable in conditions that resemble tidal breathing, once operator training was assured. The procedure adds further to the current standard assessment of breathability and allows estimating the maximal added respiratory load, about 10-20% of the respiratory resistance reported in heathy adult subjects.

Impact of Inhaled Corticosteroids on the Modulation of Respiratory Defensive Reflexes During Artificial Limb Exercise in Ovalbumin-Sensitized Rabbits.

INTRODUCTION: Cough is a major lower airway defense mechanism that can be triggered by exercise in asthma patients. Studies on cough reflex in experimental animal models revealed a decrease of cough reflex sensitivity during exercise in healthy animals, but a lack of desensitization in ovalbumin-sensitized rabbits. The aim of our study is to evaluate the impact of inhaled corticosteroids on cough reflex during artificial limb exercise in an animal model of eosinophilic airway inflammation. MATERIALS AND METHODS: Sixteen adult ovalbumin-sensitized rabbits were randomly divided into two groups. The "OVA-Corticoid" group (n = 8) received inhaled corticosteroids (budesonide; 1 mg/day during 2 consecutive days) while the "OVA-Control" (n = 8) group was exposed to saline nebulization. The sensitivity of defensive reflexes induced by direct mechanical stimulation of the trachea was studied in anesthetized animals, at rest and during artificial limb exercise. Cell count was performed on bronchoalveolar lavage fluid and middle lobe tissue sections to assess the level of eosinophilic inflammation. RESULTS: All rabbits were significantly sensitized but there was no difference in eosinophilic inflammation on bronchoalveolar lavage or tissue sections between the two groups. Artificial limb exercise resulted in a significant (p = 0.002) increase in minute ventilation by 30% (+ 209 mL.min-1, ± 102 mL/min-1), with no difference between the two groups. 322 mechanical tracheal stimulations were performed, 131 during exercise (40.7%) and 191 at rest (59.3%). Cough reflex was the main response encountered (46.9%), with a significant increase in cough reflex threshold during artificial limb exercise in the "OVA-Corticoid" group (p = 0.039). Cough reflex threshold remained unchanged in the "OVA-Control" group (p = 0.109). CONCLUSION: Inhaled corticosteroids are able to restore desensitization of the cough reflex during artificial limb exercise in an animal model of airway eosinophilic inflammation. Airway inflammation thus appears to be involved in the physiopathology of exercise-induced cough in this ovalbumin sensitized rabbit model. Inhaled anti-inflammatory treatments could have potential benefit for the management of exercise-induced cough in asthma patients.

Desensitization of the Cough Reflex Induced by Corticosteroids in Ovalbumin-Sensitized Rabbits During Artificial Limb Exercise.

INTRODUCTION: Cough is a major symptom frequently experienced during exercise, mainly in asthmatic patients. Inhaled glucocorticoids represent the keystone treatment in the management of asthma, but little is known about interactions between cough and exercise, especially in controlled patients. During exercise, cough reflex (CR) appears downregulated in healthy animal models whereas a lack of desensitization of CR has been shown in ovalbumin-sensitized animal models, mimicking asthmatic disease. AIMS AND OBJECTIVES: The goal of our study was to clarify the potential modulation of the CR induced by inhaled corticosteroids (CS) in ovalbumin (OVA) sensitized rabbits during artificial limb exercise. MATERIALS AND METHODS: Seventeen OVA sensitized rabbits were studied. Among them, 9 were treated with CS delivered intravenously (OVA-Corticoids). The ventilatory response to direct tracheal stimulation, performed at rest and during exercise, was determined to assess the incidence and the sensitivity of the CR. Broncho-alveolar lavage (BAL) and cell counts were performed to determine the level of airway inflammation. Exercise was mimicked by electrically induced hindlimb muscular contractions (EMC). RESULTS: Compared to rest values, EMC increased minute ventilation by 28% without any decrease in respiratory resistance (Rsr). Among 322 tracheal stimulations, 172 (53%) were performed at rest and 150 (47%) during exercise. The sensitivity of CR decreased during artificial limb exercise compared to baseline in OVA-Corticoids rabbits (p = 0.0313) while it remained unchanged in OVA rabbits (p = NS). CONCLUSION: Corticosteroids appear to restore the desensitization of the CR in OVA sensitized rabbits during artificial limb exercise, suggesting the potential role of airway inflammation in the pathophysiology of cough during exercise in asthmatics.

Modulation of protective reflex cough by acute immune driven inflammation of lower airways in anesthetized rabbits.

Chronic irritating cough in patients with allergic disorders may reflect behavioral or reflex response that is inappropriately matched to the stimulus present in the respiratory tract. Such dysregulated response is likely caused by sensory nerve damage driven by allergic mediators leading to cough hypersensitivity. Some indirect findings suggest that even acid-sensitive, capsaicin-insensitive A-δ fibers called "cough receptors" that are likely responsible for protective reflex cough may be modulated through immune driven inflammation. The aim of this study was to find out whether protective reflex cough is altered during acute allergic airway inflammation in rabbits sensitized to ovalbumin. In order to evaluate the effect of such inflammation exclusively on protective reflex cough, C-fiber mediated cough was silenced using general anesthesia. Cough provocation using citric acid inhalation and mechanical stimulation of trachea was realized in 16 ovalbumin (OVA) sensitized, anesthetized and tracheotomised rabbits 24h after OVA (OVA group, n = 9) or saline challenge (control group, n = 7). Number of coughs provoked by citric acid inhalation did not differ between OVA and control group (12,2 ±6,1 vs. 17,9 ± 6,9; p = 0.5). Allergic airway inflammation induced significant modulation of cough threshold (CT) to mechanical stimulus. Mechanically induced cough reflex in OVA group was either up-regulated (subgroup named "responders" CT: 50 msec (50-50); n = 5 p = 0.003) or down-regulated (subgroup named "non responders", CT: 1200 msec (1200-1200); n = 4 p = 0.001) when compared to control group (CT: 150 msec (75-525)). These results advocate that allergen may induce longer lasting changes of reflex cough pathway, leading to its up- or down-regulation. These findings may be of interest as they suggest that effective therapies for chronic cough in allergic patients should target sensitized component of both, reflex and behavioral cough.

A Calibration Device to Compare Body Plethysmographs Among Pediatric Lung Function Laboratories.

Multi-center studies in specific airway resistance have shown significant inter laboratory variability. Comparison of plethysmographic equipment using a lung model easily transportable from one site to another should be of help to international normative studies. A resistor made of parallel capillary tubes - insuring adequate linearity within 1 L/sec - was connected to a glass bottle. Thermal time constants were measured while the bottle was empty and while stuffed with steel wool. In the latter, isothermal condition was estimated to occur only at very low frequency (around 0.01 Hz) and gas compression was polytropic up to 0.6 Hz. With the empty analog, adiabatic gas compression was estimated to occur at frequencies ≥0.2 Hz, and more accurate volume estimation was obtained. The empty analog volume and specific resistance measured in a body plethysmograph on different days indicated within 5% accuracy as well as intersession repeatability. It is concluded that a physical analog built out of simple material provides accurate measurements of specific resistance. The apparatus should be of help to compare plethysmographic equipments from different laboratories.

Modulation of defensive airway reflexes during continuous positive airway pressure in the rabbit.

Lung hyperinflation may alter the pattern of ventilatory reflexes in chronic respiratory disorders. The aim of the study was to test the effect of continuous positive airway pressure (CPAP) on ventilatory responses to mechanical stimulation of the trachea.

Down-Regulation of Cough during Exercise Is Less Frequent in Healthy Children than Adults. Role of the Development and/or Atopy?

Cough is typically associated with physical activity in children with asthma, but the characteristics of the relationship between cough and exercise has not been established under physiological conditions. The aim of the study was to describe the effect of exercise on the reflex cough response elicited by a single breath of capsaicin in non-asthmatic children. A group of non-asthmatic adults was studied as reference. Thirty children and 29 adults were recruited. The cough reflex sensitivity to capsaicin was first determined to establish the dose that provokes 5 cough efforts (C5). The number of coughs elicited by C5 (NC5) was then compared at baseline and during a standardized submaximal treadmill exercise. Data are expressed as median (interquartile range). Children and adults showed a significant decrease in NC5 (respectively from 5.0 (4.0-6.0) to 2.5 (2.0-4.0), p < 0.0005 and from 6.0 (5.0-7.0) to 2.0 (0.0-3.0, p < 0.0005). During exercise, NC5 was observed to decrease in all adult subjects, but in only 24/30 children (80%, p = 0.02). A trend for a higher incidence of personal and familial atopy was observed in children that lacked cough down-regulation during exercise compared with other children. It is concluded that the cough reflex response to capsaicin is down regulated by exercise in both children and adults. The effect however is less consistently observed in the former. The difference may reflect maturation of descending inhibitory pathways of the cough reflex, but may also be associated to atopy. The data stress the importance of assessing the time relationship of cough and exercise in questionnaire studies of asthma.

Epinephrine but not vasopressin attenuates the airway response to anaphylactic shock in rats.

PURPOSE: The two life-threatening signs of anaphylactic shock (AS) are severe arterial hypotension and bronchospasm. Guidelines recommend epinephrine as first-line treatment. Arginine vasopressin (AVP) has been proposed as an alternative if epinephrine does not correct arterial hypotension. These two drugs may have beneficial, neutral or deleterious effects on airflow either directly or by modifying factors that regulate vasodilatation and/or edema in the bronchial wall. AIM OF THE STUDY: To compare the effects of epinephrine and AVP on airflow and airway leakage in a rat model of AS. MATERIALS AND METHODS: Thirty-two ovalbumin-sensitized rats were randomized into four groups: control (CON), AS without treatment (OVA), AS treated with epinephrine (EPI), and AS treated with AVP (AVP). Mean arterial pressure (MAP), respiratory resistance and elastance and microvascular leakage in the airways were measured. RESULTS: All OVA rats died within 20 minutes following ovalbumin injection. Ovalbumin induced severe arterial hypotension and airway obstruction (221 ± 36 hPa.s.L-1 vs. vehicle 52 ± 8 hPa.s.L-1; p < 0.0001) associated with microvascular leakage distributed throughout the trachea, bronchi and intra-pulmonary airways. EPI and AVP extended survival time; EPI restored a higher level of MAP than AVP. Airway obstruction was attenuated by epinephrine (146 ± 19 hPa.s.L-1; p < 0.0001), but not by AVP (235 ± 58 hPa.s.L-1; p = 0.42). CONCLUSIONS: Epinephrine was superior to AVP for alleviating the airway response in a rat model of AS. When bronchospasm and severe arterial hypotension are present during AS, epinephrine should be the drug of choice.

Maturation of Airway Defensive Reflexes Is Related to Development of Feeding Behavior during Growth in Rabbits.

Introduction: Cough and expiration reflex are major lower airway defense mechanisms that have not been studied throughout development in relation with the feeding behavior. Aim: To describe airway defense reflexes evoked by mechanical stimulation of the trachea in developing rabbit pups. Material and Methods: Sixty one pups were allocated to 3 groups according to their feeding behavior: suckling (n = 22), weanling (n = 21) and weaning (n = 18) group. The incidence and sensitivity of defense reflexes triggered by mechanical tracheal stimulation were studied in anesthetized and tracheotomized animals. Data are expressed as median (25th to 75th percentile). Results: The overall incidence of defensive responses (cough and/or expiration reflex) was found to be significantly higher in suckling [100% (50-100%); p = 0.01] and weanling [75% (40-100%); p = 0.05] animals when compared to weaning ones [37.5% (0-75%)]. However, cough motor pattern accounted for only 29% (0-62%) of all defensive responses in suckling rabbits and its frequency was significantly lower in this group when compared with weanling [100%(50-100%); p = 0.006] or weaning group [62%(50-100%), p = 0.05]. In other word the expiration reflex was the dominant response in suckling animals. Conclusion: Incidence and motor pattern of defensive responses were found to be linked to the pup feeding behavior and the expiration reflex was the major response triggered in suckling pups. The results suggest that this reflex is especially fitted to occur during the coordinated swallowing - breathing fast activities of sucking.

Lack of desensitization of the cough reflex in ovalbumin-sensitized rabbits during exercise.

INTRODUCTION: Cough is a major symptom of asthma frequently experienced during exercise but little is known about interactions between cough and exercise. The goal of our study was to clarify the potential modulation of the cough reflex (CR) by exercise in a spontaneously breathing anaesthetized animal model of airway eosinophilic inflammation. MATERIALS & METHODS: Ten ovalbumin (OVA) sensitized adult rabbits and 8 controls were studied. The ventilatory response to direct tracheal stimulation, performed both at rest and during exercise was determined to quantify the incidence and the sensitivity of the CR. Broncho-alveolar lavages (BAL) and cell counts were performed to assess the level of the airway inflammation following OVA-induced sensitization. Exercise was mimicked by Electrically induced hindlimb Muscular Contractions (EMC). RESULTS: Among 494 tracheal stimulations, 261 were performed at rest and 233 at exercise. OVA challenges in sensitized rabbits caused a significant increase in the percentage of eosinophils (p = 0.008) in BAL. EMC increased minute ventilation by 36% and 35% in OVA and control rabbits respectively, compared to rest values. The sensitivity of the CR decreased during exercise compared to baseline in control rabbits (p = 0.0313) while it remained unchanged in OVA rabbits. CONCLUSION: The desensitization of the CR during exercise in control rabbits was abolished in OVA rabbits. The precise role of airway inflammation in this lack of CR desensitization needs to be further investigated but it might contribute to the exercise-induced cough in asthmatics.

Breathing-related changes of respiratory resistance in vocal cord dysfunction.

Vocal cord dysfunction induced by exercise in children with uncontrolled asthma was identified by laryngoscopy. The paradoxical adduction of the vocal cords was also indicated by the breathing-related changes of the forced oscillation respiratory resistance showing prominent increase during inspiration and a large positive difference between inspiration and expiration. The breathing-related changes of respiratory resistance offer thus a useful first-line technique to diagnose vocal cord dysfunction.

Specific airway resistance in healthy young Vietnamese and Caucasian adults.

In healthy Vietnamese children the respiratory resistance has been suggested to be similar at 110 cm height but larger at 130 cm when compared with data in Caucasians from the literature, suggesting smaller airways in older Vietnamese children (Vu et al., 2008). The hypothesis tested here is whether the difference in airway resistance remains consistent throughout growth, and if it is larger in adult Vietnamese than in Caucasians. Airway resistance and Functional Residual Capacity were measured in healthy young Caucasian and Vietnamese adults in their respective native country using identical equipment and protocols. Ninety five subjects in Vietnam (60 males) and 101 in France (41 males) were recruited. Airway resistance was significantly larger in Vietnamese than in Caucasians and in females than in males, consistent with difference in body dimensions. Specific airway resistance however was not different by ethnicity or gender. The findings do not support the hypothesis that airway size at adult age - once normalized for lung volume - differs between Vietnamese and Caucasians.

Neonatal hyperoxia up regulates cough reflex in young rabbits.

A link between cough in childhood and neonatal exposure to hyperoxia following preterm birth has been suggested. The hypothesis is tested that neonatal exposure to hyperoxia is associated with long term up regulation of cough. Airway hyperresponsiveness is also investigated because it is a frequent finding in children born preterm and may be linked to cough. Thirty nine young rabbits survived a 48 h neonatal exposure to 93% oxygen (H, n=26) or continued room air (A, n=13). Cough response to mechanical tracheal stimulation and airway responses to a mock exercise and to methacholine were studied under anaesthesia. Larger incidence of coughs and increased sensitivity to tracheal stimulation occurred in H vs A (p<0.01), while bronchial responses to exercise or methacholine were similar in both groups. Neonatal hyperoxia is associated with up regulation of cough in young rabbits. The lack of a parallel increase in airway responsiveness suggests up regulation may be of central origin.

Flow dependence of specific airway resistance and diagnostic of asthma in children.

Panting majors turbulent flow and contribution of larger airways to the measurement of specific airway resistance (sRaw). The hypothesis was tested that the difference between asthmatic and healthy children is enhanced by narrowing the flow interval to compute sRaw. sRaw was measured during panting in 40 asthmatic and 25 healthy children and computed using all data points (full scale flow) and limited to the flow intervals ± 1L/sec and ± 0.5 L/sec. sRaw was not different between asthmatics (0.87 ± 0.20 kPa.s) and controls (0.80 ± 0.25 kPa.s) when computed full scale, while it was significantly larger in asthmatics than controls within ± 1L/sec (0.77 ± 0.16 kPa.s vs 0.65 ± 0.15 kPa.s, P < 0.004) or ± 0.5 L/sec (0.77 ± 0.21 kPa.s vs 0.61 ± 0.17 kPa.s, P < 0.002). On the other hand, the within subject coefficient of variation was significantly larger when sRaw was computed within ± 1L/sec (13.7 ± 7.2%) or ± 0.5 L/sec (28.3 ± 18.1%) than full scale (11.0 ± 6.7%), respectively P < 0.002 and P < 0.0001. It is concluded that narrowing the flow interval to compute sRaw is associated with better discrimination between asthma and health in children, although the short term variability of sRaw is increased.

Airway obstruction, upper airway artifact and response to bronchodilator in asthmatic and healthy children.

BACKGROUND: The forced oscillation technique (FOT) may be affected by the upper airway artifact in children with airway obstruction. The aim was to test the impact of the artifact on FOT ability to diagnose asthma in children as recommended by current guidelines. METHODS: The FOT was performed in 58 asthmatics and 27 controls before and after salbutamol. Respiratory resistance (Rrs) was measured at 8 Hz with a standard generator (SG), and a head generator (HG) that minimizes the upper airway artifact. The response to salbutamol was computed as change in respiratory resistance (Δ%Rrs) and admittance (Δ%Ars) that almost cancels the effect of the upper airway artifact. RESULTS: Rrs and Δ%Rrs were significantly larger in patients than controls by HG (respectively P < 0.001 and P = 0.002) but not SG, while Δ%Ars was larger in asthma than control by both (P < 0.04). Best discriminators between patients and controls were Rrs or Δ%Rrs by HG and Δ%Ars by SG. CONCLUSION: In asthmatic children, the upper airway artifact significantly impacts FOT measurements. The diagnostic value may be improved by minimization of the shunt, such as the computation of Δ%Ars.

Specific airway resistance in children: panting or tidal breathing?

BACKGROUND: The measurement of specific airway resistance during tidal breathing (sRaw(tb)) has gained popularity in children, but methodological concerns have been raised regarding the electronic compensation for the thermal artifact. The panting method (sRaw(p)) is efficient in minimizing the latter, but may be associated with a change in end expiratory lung volume if the effort is not properly balanced. The aim of the study was to compare sRaw(tb) with sRaw(p) in children. METHODS: Fifty-five children aged 6.5-11.5 years were studied. sRaw(tb) was measured in a commercial plethysmograph. sRaw(p) was measured with a home made equipment that allowed breath by breath analysis (sRaw(p1)) as well as with the commercial body box (sRaw(p2)). RESULTS: sRaw(tb) was significantly larger than either sRaw(p1) or sRaw(p2) (P < 0.0001). The mean (95% CI) difference sRaw(p1) - sRaw(tb) was -0.374 (-0.835 to 0.088) kPa s. The difference between sRaw(p1) and sRaw(p2) was significant (P < 0.005) but not clinically relevant, and mean (95% CI) difference sRaw(p1) - sRaw(p2) was 0.115 (-0.094 to 0.324) kPa s. The breath by breath analysis showed small but significant increase in sRaw(p1) throughout the maneuver (P < 0.001), whatever the pattern of end expiratory level. CONCLUSION: Tidal breathing is associated with an overestimation of sRaw compared with panting in children. Although the latter results in small increase throughout the panting maneuver, sRaw(p) is probably more trustful than sRaw(tb).

Desensitization of the cough reflex during limb muscle contraction in anesthetized rabbits.

The 'cough network' exhibits plasticity at the sensor and integration levels leading to modulation of the strength or pattern of the cough reflex. Little is known about the interactions between cough and human activities, especially during exercise. The present study was designed to determine whether exercise, mimicked by electrically induced muscle contractions, can modify the incidence and/or strength of cough following mechanical stimulation of the trachea in anesthetized rabbits. Thirteen anesthetized, tracheotomized rabbits were studied by a total of 311 tracheal stimulations: 196 at rest and 115 during exercise. During muscle contractions, the incidence of the cough reflex (CR) decreased and the expiration reflex (ER) increased (p < 0.0001). The sensitivity of the CR and ER both decreased during exercise compared to the sensitivity of the CR at rest (p < 0.02), while the strength of the expulsive response remained unchanged. These results indicate that adjustments occurring during muscle contractions likely downregulate tracheal defensive reflexes in anesthetized rabbits.

Nasal stimulation by water down-regulates cough in anesthetized rabbits.

The 'cough center' can be tuned by various afferent inputs, suggesting possible interactions at a central level of neural pathways originating from distant anatomical sites. The present study was designed to determine whether brief mechanical stimulation of the trachea can trigger cough during apnea elicited by nasal instillation of water. Twelve anesthetized, tracheotomized rabbits were studied. Mechanical stimulation of the trachea was performed under 3 conditions: baseline control, after instillation of saline into the nose and during apnea following instillation of water. The baseline breathing pattern did not differ between the 3 conditions. In a series of 171 stimulations, expiration reflex occurred in 81% of stimulations during apnea with a significantly (p<0.0001) lower peak expiratory flow than at baseline or during saline instillation. The incidence of responses comprising a cough reflex was also lower during water instillation than at either baseline or with saline (p<0.0001). These results indicate that stimulation of nasal afferents with distilled water likely down-regulates cough.