Nasal heat exchange in the giraffe and other large mammals.

The respiratory air of the giraffe is exhaled at temperatures substantially below body core temperature. As a consequence, the water content of the exhaled air is reduced to levels below that in pulmonary air, resulting in substantial reductions in respiratory water loss. Measurements under outdoor conditions showed that at an ambient air temperature of 24 degrees C, the exhaled air was 7 degrees C below body core temperature, and at ambient air temperature of 17 degrees C, the exhaled air was 13 degrees C below core temperature. The observations were extended to two additional species of wild and four species of domestic ungulates. All these animals exhaled air at temperatures below body core temperature. The average amount of water recovered due to cooling of the air during exhalation, calculated as per cent of the water loss that would occur if air were exhaled at body core temperature, amounted to between 24 and 58%, the average value for the giraffe being 56%.     

Respiratory heat loss during work at various ambient temperatures

The purpose of this investigation was to establish the temperature and humidity of the expired air of subjects working at various metabolic rates at ambient temperatures between -40 degrees C and 20 degrees C in order to calculate respiratory heat loss. Measurements of the respired air temperature and water vapour content were made for five subjects while they either stood or walked on a treadmill. The results indicated that the maximum respired air temperature varied slightly with the ambient air temperature but changes in metabolic rate, respiration rate and breathing frequency had no apparent effect on the expired air temperature under the conditions studied. The relative humidity of the respired air was found to be close to saturation in the extreme-cold environments. Heat loss due to respiration was between 25 and 30% of the resting metabolic and between 15 and 20% of the working metabolic rate.     

Respiratory heat loss is not the sole stimulus for bronchoconstriction induced by isocapnic hyperpnea with dry air

It is uncertain if respiratory heat loss or respiratory water loss is the stimulus for bronchoconstriction induced by isocapnic hyperpnea or exercise with dry air in subjects with asthma. We partially separated these 2 stimuli by having 18 subjects with asthma breathe dry air (0 mg/L water content) at increasing ventilations by isocapnic hyperpnea while we measured the increase in specific airway resistance (SRaw). The study was divided into 2 phases. In Phase 1, we used an apparatus with a single respiratory valve and evaluated the subjects' responses at 3 different inspired temperatures (-8.4, 20.5, and 39.4 degrees C). Seven of the subjects had esophageal catheters with 2 thermocouples in place to measure retrocardiac and retrotracheal temperatures. In this phase, we found that there were no significant differences in the ventilation required to cause a 100% increase in SRaw among the 3 different inspired temperatures (48.4 L/min, cold; 47.5 L/min, room temperature; 44.2 L/min, hot), even though the retrotracheal temperature fell more when the subjects breathed cold air at 40 L/min (2.1 degrees C) than when they breathed hot air (1.2 degrees C), suggesting greater airway cooling with the cold air. In Phase 2, in order to accurately measure inspired and exhaled temperatures and exhaled water content, we used 2 separate systems for delivering the inspired air and collecting the exhaled air at 2 different inspired temperatures (-21.4 and 38.9 degrees C). Again, we found that there was no significant difference in the ventilation required to cause a 100% increase in SRaw between the 2 different inspired temperatures (28.3 L/min, cold; 33.6 L/min, hot). When the subjects inhaled cold air, exhaled temperature was warmer than previously reported.(ABSTRACT TRUNCATED AT 250 WORDS)     

Thermoregulatory and cardiac responses of infant spontaneously hypertensive and Wistar-Kyoto rats to cold exposure.

Cardiovascular function during cold exposure is dependent on effective thermoregulation. This dependence is particularly apparent in infants. For example, we have previously demonstrated that in infant rats during cold exposure, cardiac rate is directly related to their ability to produce heat endogenously. The primary source of endogenous heat production for infant rats is brown adipose tissue (BAT). Because of the dependence of cardiac rate on effective thermoregulation in the cold and because hypertension in spontaneously hypertensive rats (SHR) is influenced by the preweanling environment, in this study we examined the thermoregulatory and cardiac rate responses of infant SHR and Wistar-Kyoto rats (WKY) to varying levels of cold exposure. In experiment 1, 7- to 8-day-old SHR and WKY were acclimated at a thermoneutral air temperature (35 degrees C) and then exposed to successive decreases in ambient temperature (30.5 degrees C, 26.5 degrees C, 23 degrees C, and 17 degrees C) while thermal and metabolic measures were recorded. Although both strains increased BAT thermogenesis and oxygen consumption in response to cold exposure, SHR cooled more than WKY and exhibited lower levels of oxygen consumption at the lowest air temperatures. Experiment 2 was identical to experiment 1 except that cardiac rate was also measured. Again, SHR exhibited substantial thermoregulatory deficits compared with WKY; in addition, they were less able than WKY to maintain cardiac rate at the 2 lowest air temperatures tested. Finally, in experiment 3, infant SHR exhibited diminished BAT thermogenesis in response to a range of doses of a selective beta3-adrenoceptor agonist. We hypothesize that long-term thermoregulatory deficits during the early postnatal period influence cardiovascular function and contribute to the development of hypertension in SHR.     

Ventilation and oxygen extraction in the little penguin (Eudyptula minor), at different temperatures in air and water

Oxygen consumption, (VO2), carbon dioxide production (VCO2) and ventilation were measured in little penguins (Eudyptula minor) in air and water over a range of ambient temperatures. Barometrically measured minimum ventilation was very low (112 +/- 23 ml.min-1 over the range 15-20 degrees C, n = 9), principally due to a lower than predicted respiratory frequency (f) (6.9 +/- 1.1 min-1). This low ventilation resulted in a very high oxygen extraction (EO2) and low air convection requirement (VI/VO2). Despite large increases in metabolic rate in birds in cold water, VI/VO2 and hence EO2 did not vary in non-heat exposed penguins over a range of air and water temperatures. Mean extraction was 53.2 +/- 10.8% (n = 82) corresponding to an air convection requirement of 11.3 +/- 2.5 L.L-1. Above thermoneutrality there was a dramatic increase in ventilation and fall in EO2 resulting from increases in both f and tidal volume (VT). End-expired gases were measured at thermoneutrality and during heat exposure. At thermoneutrality FE'CO2 was 0.074 +/- 0.005, and FE'O2 0.115 +/- 0.009. During heat exposure end expired gases approached atmospheric levels.     

The design and the role of the nasal passages in temperature regulation in the dik-dik antelope (Rhynchotragus kirkii) with observations on the carotid rete

Exhaled air temperatures (Tex) and rectal temperatures (Trec) were measured in five dik-dik antelopes under controlled environmental temperatures (Ta) between 15 and 40 degrees C. In addition morphometric analysis of the nasal passages, gross and histological identification of the carotid rete were carried out in an attempt to illustrate a possible brain cooling system in this small ungulate. Below Ta of 30 degrees C, Tex decreased with decreasing Ta leading to the establishment of a temperature gradient of about 20 degrees C between Ta and Trec at a Ta of around 15 degrees C. At Ta of 30 degrees C Tex was approximately half a degree lower than the Trec. Gross and histological studies revealed the presence of carotid rete and profuse vascularisation of the nasal turbinates. Morphometric analysis established a mean volume density of the nasal passages (np) in the splanchnocranium (sp) (VV(np,sp], surface density (SV(np,sp] and harmonic mean width of the nasal passages (W) of 9.5%, 2.15 cm2/cm3 and 1.23 mm, respectively. Combined physiological and anatomical measurements suggest that the dik-dik may possess an efficient brain cooling and water conserving system. The design of the upper respiratory system in these antelopes may help these animals to tolerate the extremes of temperatures and insolation encountered in their habitat.     

Influence of ambient and ventilator output temperatures on performance of heated-wire humidifiers

Although heated humidifiers are considered the most efficient humidification devices for mechanical ventilation, endotracheal tube occlusion caused by dry secretions has been reported with heated-wire humidifiers. We tested the hypothesis that inlet chamber temperature, influenced by ambient air and ventilator output temperatures, may affect humidifier performance, as assessed by hygrometry. Hygrometry was measured with three different humidifiers under several conditions, varying ambient air temperatures (high, 28-30 degrees C; and normal, 22-24 degrees C), ventilators with different gas temperatures, and two VE levels. Clinical measurements were performed to confirm bench measurements. Humidifier performance was strongly correlated with inlet chamber temperature in both the bench (p < 0.0001, r2 = 0.93) and the clinical study. With unfavorable conditions, absolute humidity of inspired gas was much lower than recommended (approximately 20 mg H2O/L). Performance was improved by specific settings or new compensatory algorithms. Hygrometry could be evaluated from condensation on the wall chamber only when ambient air temperature was normal but not with high air temperature. An increase in inlet chamber temperature induced by high ambient temperature markedly reduces the performance of heated-wire humidifiers, leading to a risk of endotracheal tube occlusion. Such systems should be avoided in these conditions unless automatic compensation algorithms are used.     

Brain cooling and respiratory heat exchange in camels during rest and exercise

Respiration in relation to brain temperature (Tb) and body temperature (Tc) were investigated in two camels at rest and one during exercise (running at 10 km/h). The animals were subjected to natural ambient conditions (day: 25-30 degrees C, relative humidity (RH) about 65%; night: 15-20 degrees C, RH approx. 90%). They were studied when fully hydrated and during progressive dehydration by up to 15% of initial weight. At low Tc (less than 38 degrees C) Tb greater than Tc by approximately 0.2 degrees C, at higher Tc significant brain cooling was observed by as much as 1.5 degrees C during exercise. Minute ventilation (VE) and respiratory rate (f) increased with Tc such that tidal volume was constant and similar at rest and during exercise (Tc-Tb) increased linearly with f and hence VE. The cooling, dependent on turbinate heat exchange was related to certain features of the air flow pattern and f which have also been described in other large mammals.     

Metabolic heat production and cold tolerance during cold stress of different intensity of adult and aged male C57BL/6J mice

Adult and aged male C57BL/6J mice were subjected to a 3-h cold stress test at either 24°C, 18°C, 12°C, or 6°C. Body mass was measured before the test, and colonic temperature, O₂ consumption, and CO₂ production were measured during the test. The slopes of colonic temperature over time of test and the mean metabolic heat production were calculated for each animal. While adult mice had a relatively small reduction in colonic temperature during the test at all four ambient temperatures, in the aged mice ambient temperatures resulted in steeper reductions of colonic temperature. In adult mice, an increase in metabolic heat production was proportional to ambient cold. The thermogenic response of aged mice at 24°C and at 18°C was similar to adult mice, suggesting that the ability of aged mice to respond to cold by increasing heat production does not diminish with age. However, in aged mice metabolic heat production at 12°C and 6°C was significantly below that of adult mice, which indicated a reduced capacity for thermogenesis.     

Heat exchnage in the black skipjack, and the blood-gas relationship of warm-bodied fishes

The black skipjack, Euthynnus lineatus, uses a centrally located vascular heat exchanger to maintain core body temperatures warmer than ambient sea water. The heat exchanger is composed of the dorsal aorta, the posterior cardinal vein, and a large vertical rete. The dorsal aorta is embedded in the posterior cardinal vein and is completely bathed in venous blood. Skipjack hemoglobin appears similar to that of the bluefin tuna in that oxygen capacity is unaffected by changing temperature. Temperature-insensitive hemoglobin may function in warm-bodied fishes to prevent the premature dissociation of oxygen from hemoglobin as blood is warmed en route to the muscles.     

Reducing ventilatory response to carbon dioxide by breathing cold air

To study the effect of cooling of nasal receptors on breathing we had 10 normal male volunteers rebreathe through their noses 8% CO2 in oxygen at "warm" (23 to 30 degrees C) and at "cold" (-4 to 10 degrees C) temperatures. In order to further examine the effect of nasal receptors on the control of breathing, 11 subjects had their nasal response to CO2 measured at the warm temperature before and after topical nasal anesthesia. To exclude an increase in nasal resistance as the cause of the reduced response to CO2, 10 subjects had their nasal resistance measured before and after nasal rebreathing of cold 8% CO2 in oxygen. To also exclude increased bronchial resistance, forced expiratory volume in one second (FEV1) was measured in 12 subjects before and after nasal breathing of cold oxygen for 3 min. The mean ventilatory response to CO2 was reduced from 3.0 +/- 1.6 L/min/mmHg to 2.5 +/- 1.1 L/min/mmHg (p less than 0.05) by the cold air. Topical nasal anesthesia increased the response to CO2 at the warm temperature from 2.4 +/- 0.7 to 2.7 +/- 0.9 L/min/mmHg. The effect of nasal breathing of 8% CO2 in oxygen at the cold temperature was to reduce nasal inspiratory resistance at 1 L/s from 4.3 +/- 3.0 cm H2O L/s to 2.6 +/- 1.0 cm H2O L/s (p less than 0.05). Expiratory resistance at 1 L/s fell from 3.7 +/- 1.5 cm H2O L/s to 2.4 +/- 0.7 cm H2O L/s (p less than 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)     

Respiratory effects of photochemical oxidant air pollution in exercising adolescents

Healthy volunteers 12 to 15 yr of age (46 boys, 13 girls) were exposed to purified air and to smoggy Los Angeles ambient air on different occasions. The studies were performed in random order approximately 2 wk apart. They included 1 h of continuous bicycle exercise (mean ventilation, 32 L/min) plus brief warm-up and cool-down periods. Symptoms and forced expiratory performance were recorded preexposure in purified air, immediately postexposure, and after 1 h recovery in purified air. Mean exposure temperature was 32 degrees C, and mean relative humidity was 45%. In ambient exposure, pollutant concentrations averaged 0.144 ppm for ozone and 153 micrograms/m3 for total suspended particulates. Group mean FEV1 decreased during ambient exposure (p less than 0.01) and only partially recovered during the following 1 h. Unlike adults studied previously, this subject group reported no significant increase in respiratory symptoms accompanying changes in FEV1. Adolescents may be less aware of early respiratory irritation by oxidants and thus more at risk from ambient exposures than are adults.     

Breathing pattern affects airway wall temperature during cold air hyperpnea in humans

We studied the influence of flow rate on respiratory heat exchange in 9 healthy adult subjects using a new noninvasive technique, the single-breath temperature washout (SBTW) curve. The SBTW curve is a plot of exhaled gas temperature versus exhaled volume during a standard exhalation and consists of an initial rise (within the first 200 ml) to a plateau temperature that persists through the remainder of exhalation. We found that exhaled gas temperatures within the initial expirate were colder at every airway locus than corresponding intra-airway gas temperatures at end-inspiration, suggesting that heat exchange occurs between lumenal gas and the relatively cooler airway walls during exhalation. The SBTW plateau temperatures were: (1) lower after preconditioning the airways with rapid (80 L/min) isocapnic hyperpnea of frigid air than after less rapid (40 L/min) cold-air hyperpnea or after quiet breathing; (2) lower when, after identical airway preconditioning regimens, the SBTW exhalation was performed with a slower (0.5 versus 2.5 L/s) expiratory flow; and (3) lower when SBTW curves were obtained after airway preconditioning using respiratory patterns with larger inspiration-expiration duration (I:E) ratios (5:1 versus 1:5) at fixed minute ventilation and respiratory rate. Our results indicate that the global respiratory gas-wall heat transfer coefficient increases with velocity to the 0.9 power, a finding similar to that in previous studies of turbulent flow in rigid pipes.     

Effects of immersion in cool water on lung-exhaled nitric oxide at rest and during exercise.

Lung nitric oxide (NO) has been postulated to relax airway and vascular smooth muscle at rest and during exercise. As a cold environment is a common cause of respiratory distress, lung exhaled NO was measured during skin and core body cooling at rest and during a progressive cycle exercise. Ten healthy male subjects were immersed in water at a water temperature (Tw) which was thermal neutral (35 degrees C) at 30 degrees C Tw, at which only skin temperature is decreased; and at 20 degrees C Tw, at which the core temperature is decreased (0.05 degrees C). At rest, V(O), and V(E) increased while exhaled NO concentration [NO] and the rate of expiration of NO (V(NO)) decreased with decreased Tw. V(O2) and ventilation (V(E)) increased with workload (W) and the values at all Tw were not different, whereas, [NO] decreased with W and the values during exercise were progressively less at all Ws as Tw declined. These results indicate that lung NO output is reduced in a graded fashion during body cooling at rest and during exercise. This suggests that lower lung NO may contribute to airway obstruction in cold environments and NO may contribute to regulation of lung heat and water exchange.     

The effect of ambient temperature on glucose tolerance.

Standard (75 g) oral glucose tolerance tests were performed at two different ambient temperatures (23 and 33 degrees C) in random order in 16 (eight obese) diabetic and 16 (eight obese) non-diabetic Nigerian subjects. Consistently higher plasma glucose values were found 120 min post-glucose ingestion at 33 degrees C, with mean differences of 0.5 (SE 0.3) and 4.5 (SE 1.5) mmol l-1 in the non-diabetic and diabetic subjects, respectively. This caused reclassification of two of the non-diabetic subjects as Impaired Glucose Tolerance at 33 degrees C, applying the WHO criteria. The difference was consistently greater (p less than 0.01) in the non-obese subjects (non-diabetic 0.8 (0.4), diabetic 6.9 (2.8) mmol l-1) than in the obese (non-diabetic 0.2 (0.4), diabetic 2.1 (0.9) mmol l-1). In the diabetic subjects, a negative correlation (r = -0.50, p less than 0.01) was established between the difference and the body mass index. This variability in responses to ambient temperature between the obese and non-obese subjects could be due to a variable influence of heat on arm blood flow consequent on differences in amounts of subcutaneous fat. The ambient temperature for the conduct of the oral glucose tolerance test is important.     

Effects of water temperature on pulmonary volumes in immersed human subjects

Pulmonary volumes and capacities have been measured at three water temperatures (Tw = 25, 34, 40 degrees C) in standing subjects immersed up to the shoulders. The comparison of data obtained in air with those obtained in thermoneutral immersion (Tw = 34 degrees C) confirms the results previously published in several studies. The comparison of data obtained in immersion at different Tw shows: 1. A significant decrease in vital capacity (VC) with bath temperature (VC 40 degrees C greater than VC 34 degrees C greater than VC 25 degrees C). The same decrease is observed in the inspiratory reserve volume (IRV) while the expiratory reserve volume (ERV), the residual volume (RV) and the functional residual capacity (FRC) do not vary. 2. A significant decrease in maximum breathing capacity (MBC) with bath temperature (MBC 40 degrees C greater than MBC 25 degrees C). 3. A significant increase in tidal volume (VT) in cold or hot water compared to thermoneutral water (VT40 degrees C greater than VT34 degrees C; VT34 degrees C less than VT25 degrees C) during quiet breathing. Breathing frequency does not change, thus ventilation (V) follows the same evolution as VT. The relative abdominal (ABD) contribution to VT, estimated by a double belt inductance plethysmograph, is reduced at Tw = 25 degrees C but unchanged at Tw = 40 degrees C compared to thermoneutral bath. Beside variations in the metabolic state, the variations of the pulmonary volumes as a function of Tw are estimated to be mainly due to alterations in respiratory muscles functioning.     

Energy turnover and heat exchange in mature lean and obese Zucker rats acutely exposed to three environmental temperatures for 24 hours

Differences between lean (FA/?, n = 6) and obese (fa/fa, n = 6) mature male Zucker rats' energy turnover and heat storage were compared during a 24-h period when the animals were exposed to ambient temperatures of 30, 15 or 5 degrees C. Energy turnover was examined through measurements of heat production rates via indirect calorimetry and heat loss rates via direct calorimetry. Heat storage rates were calculated as the difference between heat production and heat loss rates. Predicted heat storage rates were also calculated as the product of the change in core temperature and the calculated specific heat of the animal based on body composition (carcass) analysis. A minimal heat loss rate was determined for each animal representing a period of least activity. Various comparisons were made: between groups (lean/obese), temperature (30, 15, 5 degrees C), calorimetry method (indirect/direct), period (light/dark), heat storage (experimental/predicted), and minimal heat loss. Immediately before a test, pretest weight and colonic temperature were obtained. Then, the animal was placed into the calorimeter chamber and remained there unrestrained for 24 h. Normal light/dark periods were maintained. On removal from the calorimeter, core temperature and body weight measurements were again obtained. Upon completion of all tests, body composition was analyzed and surface area determined. Energy turnover, i.e. both heat production and heat loss in the lean and obese animals differed among the 30, 15 and 5 degrees C exposures. The obese animals had relatively greater heat production rate and heat loss rate (kcal/day or kcal/kg (FFM)/day than the lean animals at 30, 15 and 5 degrees C. But, on a relative basis, the increments in heat production in the cold environments were greater for the lean animals. Both the lean and obese animals tended to be more active during the dark period when at 30 degrees C, but the difference was less at 15 degrees C and even less at 5 degrees C. Experimental heat storage rates did not differ significantly from predicted values at any of the temperatures with the possible exception of the animals at 5 degrees C. It was concluded that the mature obese Zucker rats had no major discernible defect in thermoregulation as revealed by rates of heat production and loss, although three of the obese rats did elicit a drop in colonic temperature during exposure to 5 degrees C, i.e. their excessive subcutaneous adiposity and thermal insulation did not prevent a fall in colonic temperature.(ABSTRACT TRUNCATED AT 400 WORDS)     

Evidence for a hormonal tactic maximizing green turtle reproduction in response to a pervasive ecological stressor

Mortality of breeding sea turtles due to excessive heat exposure after nesting activities is an unusual feature of the Raine Island green turtle rookery. Breeding turtles that fail to return to the ocean after oviposition can experience increasing body temperatures that exceed lethal limits (>39 degrees C) as ambient temperatures rise after sunrise. We investigated how acute increases in body temperature influenced plasma corticosterone (B) concentrations of individual turtles. Furthermore, interactions between progesterone (P) and testosterone (T) and increasing body temperature and the glucocorticoid corticosterone were examined for negative correlations. Breeding green turtles exhibited a 16-fold mean increase in plasma corticosterone concentration as body temperature (cloacal) rose from 28.2 to 40.7 degrees C in less than 6 h. However, the absolute increase in plasma B was small and much less than expected, despite the lethal stressor. Comparatively, the maximal B response to lethal heat stress was similar to plasma B concentrations obtained from breeding female turtles exposed to 8 h of capture stress. However, the maximal B response of breeding turtles exposed to heat and capture stressors was significantly less than the B response of nonbreeding adult female turtles subjected to an 8-h capture stressor. No negative correlations were observed between plasma T and plasma B, between plasma T and body temperature, between plasma P and plasma B, or between plasma P and body temperature. Our findings provide further evidence that reduced adrenocortical function operates in breeding green turtles in the presence of even the most pervasive of environmental stressors.     

Effect of atmospheric nitric oxide (NO) on measurements of exhaled NO in asthmatic children

The measurement of exhaled nitric oxide concentrations [NO] may provide a simple, noninvasive means for measuring airway inflammation. However, several measurement conditions may influence exhaled NO levels, and ambient NO may be one of these. We measured exhaled NO levels in 47 stable asthmatic children age 5 to 17 years and in 47 healthy children, gender and age matched. Exhaled [NO] in expired air was measured by a tidal breathing method with a chemiluminescence analyzer, sampling at the expiratory side of the mouthpiece. NO steady-state levels were recorded. In order to keep the soft palate closed and avoid nasal contamination, the breathing circuit had a restrictor providing an expiratory pressure of 3–4 cm H₂O at the mouthpiece. To evaluate the effect of [NO] in ambient air, measurements were randomly performed by breathing ambient air or NO-free air from a closed circuit. Breathing NO-free air, exhaled [NO] in asthmatics (mean ± SEM) was 23.7 ± 1.4 ppb, significantly higher (P < 0.001) than in healthy controls (8.7 ± 0.4 ppb). Exhaled NO concentrations measured during ambient air breathing were higher (49 ± 4.6 ppb, P < 0.001) than when breathing NO-free air (23.7 ± 1.4 ppb) and were significantly correlated (r = 0.89, P < 0.001) with atmospheric concentrations of NO (range 3–430 ppb). These findings show that (1) exhaled [NO] values of asthmatic children are significantly higher than in healthy controls, and (2) atmospheric NO levels critically influence the measurement of exhaled [NO]. Therefore, using a tidal breathing method the inhalation of NO-free air during the test is recommended. Pediatr Pulmonol. 1998; 26:30–34. © 1998 Wiley-Liss, Inc.     

Characterization of exhaled nitric oxide: introducing a new reproducible method for nasal nitric oxide measurements.

Nitric oxide (NO) is present in the human nasal airways and has been suggested to originate primarily from the paranasal sinuses. The aim of this study was to establish a new and reproducible method for measurement of nasal NO. Through repeated single-breath measurements the intra- and inter-individual variations of NO levels in nasally (into a tightly fitting mask covering the nose) and orally exhaled air were determined in healthy humans. Variations due to the methods used were investigated. The contribution of oral NO to the nasal exhalations by introducing a mouthwash procedure was also studied. This study shows distinct individual values of NO in nasally and orally exhaled air of healthy humans. Some diurnal variability was also found with a rise in NO in nasally and orally exhaled air over the day, but no, or little, day-to-day variability when comparing the results from separate mornings. There was no correlation between NO levels in nasally and orally exhaled air, whereas there was a strong correlation between NO levels in air exhaled through the left and right nostril. The levels of NO in air exhaled at 0.17 L x s(-1) through either nostril separately were higher than in air exhaled at the same flow rate through both nostrils simultaneously. After the introduction of a mouthwash procedure the level of NO in orally, but not nasally exhaled air was reduced. To conclude the method using nasal exhalation into a nose mask is highly reproducible. It is also suggested that subtracting the level of NO in orally exhaled air, after mouthwash, from that in nasally exhaled air, would adequately reflect nasal NO levels.