Bradykinin B1 receptor up-regulation by interleukin-1beta and B1 agonist occurs through independent and synergistic intracellular signaling mechanisms in human lung fibroblasts.

Bradykinin B1 receptors (B1R) are rapidly induced after tissue trauma and are thought to be involved in maintaining the inflammatory response. Little is known about the intracellular signaling pathways mediating B1R induction in response to stress and inflammation. Here, we show that up-regulation of B1R by B1R agonist and interleukin-1beta (IL-1beta) occur through distinct but synergistic pathways in IMR-90 human lung fibroblasts. Incubation of cells with the B1R agonist desArg10kallidin (desArg10KD; 100 nM) and IL-1beta (500 pg/ml) resulted in a 3- and 4-fold increase, respectively, in B1R by 6 h, whereas coincubation of these factors produced up to a 20-fold increase. Furthermore, coincubation increased the potency of IL-1beta by 2-fold. Both the individual and the synergistic responses were sensitive to genistein, a general tyrosine kinase inhibitor. On the other hand, only the desArg10KD response and the synergistic response were sensitive to the p38 mitogen-activated protein kinase inhibitor SB 203580. Furthermore, only the synergistic response was sensitive to the nuclear factor-kappaB inhibitor pyrrolidine dithiocarbamate. Despite B1R up-regulation in A549 human lung epithelial cells by desArg10KD or IL-1beta individually, these factors did not act synergistically in this cell line. In conclusion, our results reinforce the view that kinins act in concert with proinflammatory cytokines to enhance selectively the inflammatory response of certain lung cells to kinins through distinct but synergistic intracellular signaling mechanisms. Thus, kinins may exert a pivotal role in maintaining and modulating feed-forward inflammatory processes in the lung.

Regulation of bradykinin receptor gene expression in human lung fibroblasts.

In WI-38 human fibroblasts, interleukin-1 beta and tumour necrosis factor-alpha (TNF-alpha) increased bradykinin B(1) receptor mRNA, which peaked between 2 and 4 h, remaining elevated for 20 h. Binding of the bradykinin B(1) receptor selective ligand [3H]des-Arg(10)-kallidin, also increased, peaking at 4 h and remaining elevated for 20 h. The B(max) value for [3H]des-Arg(10)-kallidin rose from 280+/-102 fmol/mg (n=3) to 701+/-147 fmol/mg (n=3), but the K(D) value remained unaltered (control, 1.04+/-0.33 nM (n=3); interleukin-1 beta, 0.88+/-0.41 nM (n=3)). The interleukin-1 beta-induced [3H]des-Arg(10)-kallidin binding sites were functional receptors, as bradykinin B(1) receptor agonist-induced responses increased in treated cells. Bradykinin B(2) receptor mRNA and [3H]bradykinin binding were upregulated by interleukin-1 beta, but not TNF-alpha. The effect of interleukin-1 beta on bradykinin B(2) receptors was smaller than for bradykinin B(1) receptors. Cycloheximide prevented interleukin-1 beta-mediated increases in B(1) and B(2) binding, but not mRNA suggesting that de novo synthesis of a transcriptional activator was unnecessary.

Bradyzide, a potent non-peptide B(2) bradykinin receptor antagonist with long-lasting oral activity in animal models of inflammatory hyperalgesia.

Bradyzide is from a novel class of rodent-selective non-peptide B(2) bradykinin antagonists (1-(2-Nitrophenyl)thiosemicarbazides). Bradyzide has high affinity for the rodent B(2) receptor, displacing [(3)H]-bradykinin binding in NG108-15 cells and in Cos-7 cells expressing the rat receptor with K(I) values of 0.51+/-0.18 nM (n=3) and 0.89+/-0.27 nM (n=3), respectively. Bradyzide is a competitive antagonist, inhibiting B(2) receptor-induced (45)Ca efflux from NG108-15 cells with a pK(B) of 8.0+/-0.16 (n=5) and a Schild slope of 1.05. In the rat spinal cord and tail preparation, bradyzide inhibits bradykinin-induced ventral root depolarizations (IC(50) value; 1.6+/-0.05 nM (n=3)). Bradyzide is much less potent at the human than at the rodent B(2) receptor, displacing [(3)H]-bradykinin binding in human fibroblasts and in Cos-7 cells expressing the human B(2) receptor with K(I) values of 393+/-90 nM (n=3) and 772+/-144 nM (n=3), respectively. Bradyzide inhibits bradykinin-induced [(3)H]-inositol trisphosphate (IP(3)) formation with IC(50) values of 11.6+/-1.4 nM (n=3) at the rat and 2.4+/-0.3 microM (n=3) at the human receptor. Bradyzide does not interact with a range of other receptors, including human and rat B(1) bradykinin receptors. Bradyzide is orally available and blocks bradykinin-induced hypotension and plasma extravasation. Bradyzide shows long-lasting oral activity in rodent models of inflammatory hyperalgesia, reversing Freund's complete adjuvant (FCA)-induced mechanical hyperalgesia in the rat knee joint (ED(50), 0.84 micromol kg(-1); duration of action >4 h). It is equipotent with morphine and diclofenac, and 1000 times more potent than paracetamol, its maximal effect exceeding that of the non-steroidal anti-inflammatory drugs (NSAIDs). Bradyzide does not exhibit tolerance when administered over 6 days. In summary, bradyzide is a potent, orally active, antagonist of the B(2) bradykinin receptor, with selectivity for the rodent over the human receptor. British Journal of Pharmacology (2000) 129, 77 - 86

Autoregulation of bradykinin receptors: agonists in the presence of interleukin-1beta shift the repertoire of receptor subtypes from B2 to B1 in human lung fibroblasts.

Elevated formation of bradykinin (BK) and Lys-BK or kallidin (KD) and their carboxypeptidase metabolites desArg(9)BK and desArg(10)KD is evident at sites of inflammation. Moreover, B2 receptors (B2R), which mediate the action of BK and KD, participates in the acute stage of the inflammatory and pain response, whereas B1 receptors (B1R), through which desArg(9)BK and desArg(10)KD act, partake in the chronic stage. We hypothesized that kinins autoregulate B2R and B1R expression in favor of B1R. Incubation of IMR-90 cells with BK (100 nM) led to a loss (89%) of B2R with a half-life (T(1/2)) of 7.0 min. Concomitantly, BK increased B1R (2- to 3-fold) with a T(1/2) of 120 min. DesArg(10)KD (100 nM) had no effect on B2R but increased B1R (3- to 4-fold) with the same rate as BK. Interleukin-1beta (IL-1beta; 500 pg/ml) also increased B1R (4- to 6-fold). Although both desArg(10)KD and BK increased the level of IL-1beta mRNA, IL-1beta receptor antagonist inhibited the increase in B1R only in response to BK. DesArg(10)KD and BK synergistically increased B1R (9-fold), which was further increased by inclusion of IL-1beta (36-fold). Therefore, kinin metabolism and kinin-stimulated production of cytokines may play a pivotal role in shifting the repertoire of kinin receptor subtypes in favor of B1R during inflammation.

Use of nasal peak flow to assess nasal patency.

Nasal patency is usually assessed in the laboratory by measuring nasal airflow conductance (Gnaw); peak inspiratory and/or expiratory flow measurements via the nose (PIFna, PEFna) have been proposed as simple alternatives suitable for home monitoring of rhinitis. We have compared the scale of changes in PIFna and PEFna (measured with a pneumotachograph) with changes in Gnaw (measured by the forced-oscillation technique) when nasal patency was increased by a topical alpha-adrenergic agonist, xylometazoline (five control subjects, seven with seasonal rhinitis, studied when asymptomatic) or decreased by topical histamine (eight control subjects). In further experiments, we altered intrapulmonary airway calibre by having subjects inhale histamine or salbutamol aerosols and examined effects on the configuration of nasal flow-volume curves (six subjects with rhinitis and mild asthma). After topical xylometazoline, there was a mean 283% increase in Gnaw, 80% increase in PEFna, and 63% increase in PIFna. After topical histamine, there was a mean 72% decrease in Gnaw, 38% decrease in PEFna, and 39% decrease in PIFna. Inducing intrapulmonary airway obstruction sometimes obscured changes in nasal patency by removing the effects of added nasal resistance on expiration and preventing development of flow limitation in the nose on inspiration. Thus, after topical drug treatment to the nose, changes in Gnaw were considerably larger than in PEFna or PIFna, which were proportionately similar. Because PIFna is usually restricted by nasal flow limitation, it is probably superior to PEFna for assessing nasal patency. When effort is submaximal, intrapulmonary dynamic resistance is increased, or nasal dynamic resistance is low, PEFna and PIFna can give a misleading impression of nasal patency. These errors can be avoided by comparisons with mouth PEF and/or PIF, suggesting that nasal and mouth peak flow should both be measured during home monitoring.

Selective labelling of bradykinin receptor subtypes in WI38 human lung fibroblasts.

1. Binding of the B1 bradykinin receptor radioligand, [3H]-des-Arg10-kallidin (-KD) and the B2 receptor radioligand [3H]-bradykinin (-BK) was investigated in membranes prepared from WI38 human foetal lung fibroblasts. 2. One-site analysis of the saturation data for [3H]-des-Arg10-KD gave an equilibrium dissociation constant (KD) value of 0.51 +/- 0.12 nM and a maximum receptor density (Bmax) of 260 +/- 49 fmol mg-1 of protein. [3H]-des-Arg10-KD binding was displaced by ligands in the order: des-Arg10-KD > KD > > des-Arg9[Leu8]-BK > des-Arg9-BK > Hoe 140 > > BK, implying that it was binding selectively to B1 receptors. 3. One-site analysis of the binding of [3H]-BK to W138 membranes indicated that it had a KD value of 0.25 +/- 0.06 nM and a Bmax of 753 +/- 98 fmol mg-1 of protein. The potencies for displacement of [3H]-BK binding were: Hoe 140 > > BK = KD > > > des-Arg10-KD = des-Arg9[Leu8]-BK = des-Arg9-BK, which was consistent with binding to B2 receptors. 4. This is the first characterization of [3H]-des-Arg10-KD binding to include both kinetic and equilibrium data, and demonstrates that [3H]-des-Arg10-KD has a high affinity for human B1 bradykinin receptors and is sufficiently selective to be used as a radioligand for B1 receptors in human cells or tissues expressing an excess of B2 BK receptors.

Evaluation of a new interrupter device for measuring bronchial responsiveness and the response to bronchodilator in 3 year old children.

The interrupter technique for measuring airway resistance is non-invasive and convenient, and therefore ideally suited for the assessment of induced changes in airway calibre in preschool children. The aim of this study was to evaluate a commercially available interrupter device (based on Microlab 4000), which calculates the interrupter resistance (Rint) from pressure and flow following a brief interruption of expiration during quiet breathing. The repeatability of Rint was assessed, and its response to methacholine challenge and the bronchodilator salbutamol were compared with an indirect technique, the fall in transcutaneous oxygen tension (Ptc,O2), using the sensitivity index (SI, i.e. the change after challenge expressed in multiples of the baseline standard deviation) in 12 wheezy children (aged 3 yrs +/- 2 months). The mean (SD) baseline value of Rint was 0.91 (0.20) kPa.L-1.s. Short-term repeatability and baseline variability were satisfactory for Rint (intraclass correlation coefficient = 0.6; mean intrasubject coefficient of variation = 13%). Although 10 of the 12 subjects obtained a significant response using Rint at maximal bronchoconstriction (i.e. SI > 2), overall, Rint was five times less sensitive than Ptc,O2 (geometric mean SI: Rint 3 vs Ptc,O2 16; p < 0.0001). Reversal of obstruction with administration of a bronchodilator was clearly demonstrated in almost all subjects: Rint after challenge (mean +/- SD) 1.25 (0.22) kPa.L-1.s; after salbutamol 0.78 (0.19) kPa.L-1.s; p < 0.001. In conclusion, the convenient interrupter resistance method appears more promising for detecting bronchodilator responses than induced bronchoconstriction in wheezy preschool children; however, measurement of transcutaneous oxygen tension provides a reliable indirect means of detecting induced airway obstruction in this age-group.

B1 bradykinin receptors and sensory neurones.

1. The location of the B1 bradykinin receptors involved in inflammatory hyperalgesia was investigated. 2. No specific binding of the B1 bradykinin receptor ligand [3H]-des-Arg10-kallidin was detected in primary cultures of rat dorsal root ganglion neurones, even after treatment with interleukin-1 beta (100 iu ml-1). 3. In dorsal root ganglion neurones, activation of B2 bradykinin receptors stimulated polyphosphoinositidase C. In contrast, B1 bradykinin receptor agonists (des-Arg9-bradykinin up to 10 microM and des-Arg10-kallidin up to 1 microM) failed to activate polyphosphoinositidase C, even in neurones that had been treated with interleukin-1 beta (100 iu ml-1), prostaglandin E2 (1 microM) or prostaglandin I2 (1 microM). 4. Dorsal root ganglion neurones removed from rats (both neonatal and 14 days old) that had been pretreated with inflammatory mediators (Freund's complete adjuvant, or carrageenan) failed to respond to B1 bradykinin receptor selective agonists (des-Arg9-bradykinin up to 10 microM and des-Arg10-kallidin up to 1 microM). 5. Bradykinin (25 nM to 300 nM) evoked ventral root responses when applied to peripheral receptive fields or central terminals of primary afferents in the neonatal rat spinal cord and tail preparation. In contrast, des-Arg9-bradykinin (50 nM to 500 nM) failed to evoke ventral root depolarizations in either control rats or in animals that developed inflammation following ultraviolet irradiation of the tail skin. 6. The results of the present study imply that the B1 bradykinin receptors that contribute to hypersensitivity in models of persistent inflammatory hyperalgesia are located on cells other than sensory neurones where they may be responsible for releasing mediators that sensitize or activate the nociceptors.

Measurement of intracellular calcium in cell populations loaded with aequorin: neurokinin-1 responses in U373MG cells.

Changes in intracellular calcium concentration are important in mediating a wide variety of physiological responses. Recently there has been renewed interest in the use of aequorin, a protein from jellyfish that emits light when calcium is bound, to measure calcium levels in cells. We have loaded populations of cells from the human glioma line, U373MG, with aequorin. Lysis of aequorin-loaded but not control cells with detergent resulted in a luminescence signal that was dependent on extracellular calcium. Aequorin-loaded cells responded to substance P, histamine, or the calcium ionophore, ionomycin, with an increase in luminescence. Signals in response to detergent, ionomycin, or substance P could be detected up to 48 h after cells were loaded with aequorin. Other neurokinin-1 agonists but not agonists at neurokinin-2 or neurokinin-3 receptors produced luminescence signals. Neurokinin-1 antagonists inhibited the substance P-induced signal. The aequorin-loading procedure worked well with U373MG cells but not with AR42J, CHO, IMR-90, or WI-38 cells.

Evaluation of the interrupter technique for measuring change in airway resistance in 5-year-old asthmatic children.

The interrupter technique is a noninvasive method for measuring airway resistance during quiet breathing which requires minimal subject cooperation. It, therefore, has enormous potential for use in young children unable to cooperate with conventional lung function tests. We evaluated the interrupter technique during bronchial challenge with methacholine administered by the tidal breathing method in 10 5-year-old asthmatic children. The mouth pressure/time [P mo(t)] curve obtained following brief airflow interruption during the expiratory phase of quiet breathing was analyzed to determine the interrupter resistance (Rint) using four different methods: RintC, a smooth curve fit with back-extrapolation; RintEO, calculated from the pressure change after the postinterruption oscillations had decayed (end-oscillation); RintL, two-point linear fit with back-extrapolation; and RintEI, calculated from the pressure change at the end of the period of interruption. The four Rint methods were compared for repeatability and sensitivity with the direct measurement of resistance by the forced oscillation technique (Rrs), and with an independent method of measuring the response to challenge, utilizing the change in transcutaneous oxygen tension (PtcO2). The sensitivity of the methods was defined by a sensitivity index (SI), the change after challenge expressed in multiples of the baseline standard deviation. The PtcO2 method had the lowest variability and was by far the most sensitive method (geometric mean SI 18.9), at least 1 doubling concentration more sensitive than the other techniques in every subject (P < 0.05). RintL was more sensitive than the other interrupter methods (geometric mean SI: RintL 4.2; RintC 1.0; RintEO 2.7; RintEI 3.1; P < 0.05) and similar in sensitivity to Rrs (geometric mean SI 4.6) in 7 out of 10 children in which this could be measured. We conclude that the interrupter method provides a simpler method than the oscillation technique for assessing airway obstruction in this age group.

Comparison of four methods of assessing airflow resistance before and after induced airway narrowing in normal subjects.

Four methods for assessing airflow resistance were compared in seven normal adults at baseline and after inducing airway narrowing with inhaled methacholine. Airway resistance (Raw) was measured during panting at 1-2 Hz within a body plethysmograph; total lung resistance was measured by using an esophageal balloon during quiet breathing (RLq) and with doubling of frequency while maintaining the original tidal volume; total respiratory resistance (Rrs) was measured at 6 Hz during forced oscillation applied at the airway opening, and interruption resistance (Rint) was measured at midtidal expiratory flow. Three methods of obtaining Rint after airflow interruption were compared [smooth curve fit of mouth pressure (Pm) back extrapolated to valve closure; two-point linear fit of Pm back extrapolated to 15 ms after closure; and Pm at 100 ms after valve closure]. We found similar basal median values (cmH2O.l-1.s) of Raw (1.3), RLq (1.4), RL of double resting frequency (1.9), Rrs (1.7), and smooth curve fit of Pm back extrapolated to valve closure (1.5); basal values of two-point linear fit of Pm back extrapolated to 15 ms after closure (2.4) and Pm at 100 ms after valve closure (4.4) were considerably larger. After induced airway narrowing, all methods of measuring resistance showed significant increases; these were largest with RLq (median %change of 265) and smallest with the three Rint methods (median %change of 62-72). Rint and Rrs methods had poorer sensitivity for detecting bronchoconstriction than lung resistance of Raw. Of the Rint methods, end interruption pressure was the most sensitive.(ABSTRACT TRUNCATED AT 250 WORDS)

The measurement of methacholine responsiveness in 5 year old children: three methods compared.

The aim of this study was to compare the feasibility of three techniques for measuring the response to bronchial challenge in young children: a direct airway measurement, the forced oscillation technique (FOT) for determining respiratory system resistance at 6 and 8 Hz (Rrs6 and Rrs8), and two indirect methods, the change in transcutaneous oxygen tension (PtcO2) and the detection of wheeze on auscultation of the chest. Thirty children aged 5 yrs, with a history of wheeze, and six asymptomatic controls, took part in a bronchial challenge test using methacholine administered by Wright nebulizer by the tidal-breathing method. The provocative concentration which produced a 35% increase in Rrs6 (PC35Rrs6) and a 15% decreases in PtcO2 (PC15PtcO2) were determined by interpolation, and the chest was ausculated after each dose of methacholine. The FOT was found to be unreliable in this age group: in seven children, the data were technically unsatisfactory in the presence of induced bronchoconstriction, whilst in three children, changes in Rrs were inconsistent after challenge. The use of Rrs8 did not improve the detection of positive responses. PC15PtcO2 was measurable in 29 of 30 children, and in 18 of these PC35Rrs6 was also measurable. In no subject did a significant, sustained increase in Rrs occur during challenge in the absence of a significant change in PtcO2. Wheeze was audible in only 4 of 25 (16%) of the positive and in no negative challenges. With this protocol, we found the FOT to be unreliable and the auscultation method valueless and potentially dangerous, since marked falls in PtcO2 of up to 33% sometimes occurred in the absence of wheeze.(ABSTRACT TRUNCATED AT 250 WORDS)

Accuracy and sensitivity of the interrupter technique for measuring the response to bronchial challenge in normal subjects.

The interrupter technique is a non-invasive method for measuring airway calibre. Since the calculation of interrupter resistance (Rint) is critically dependent upon the analysis of the mouth pressure/time (Pmo(t)) curve obtained after flow interruption, we wanted to assess the relative merits of four different analyses of Pmo(t) curves, obtained under basal conditions and following methacholine-induced airway narrowing, in 10 healthy adults. Four methods of analysing the Pmo(t) curves were used to calculate Rint values: RintC-a smooth curve fit with back-extrapolation; RintL-two-point linear fit with back-extrapolation; RintEO-calculated from the pressure change after the post-interruption oscillations had decayed (end-oscillation); and RintEI-calculated from the pressure change at the end of the period of interruption. The airway response measured with the four Rint methods was compared with plethysmographic airway resistance (Raw). The sensitivity of the methods was determined by calculating a sensitivity index (SI), the change in resistance after challenge expressed in multiples of baseline standard deviation. Values of RintC were similar to Raw values under all conditions. Resistance values from the remaining Rint methods significantly exceeded Raw (mean basal difference: 0.13-0.34 kPa.l-1 x s; mean difference after challenge: 0.12-0.42 kPa.l-1 x s. Raw was the most sensitive method for detecting bronchoconstriction (doubling of Raw was equivalent to SI of 10.5). Of the Rint methods, RintEI gave the highest sensitivity index (SI = 3.1), with a 42% mean change; RintC produced the greatest proportionate change after challenge (55%), but with a lower SI (2.2).(ABSTRACT TRUNCATED AT 250 WORDS)

Repeatability of methacholine challenge in asthmatic children measured by change in transcutaneous oxygen tension.

BACKGROUND: The airway response to bronchial provocation may be evaluated by monitoring the fall in transcutaneous oxygen tension (PtcO2) but the repeatability of this method has not been rigorously assessed. METHODS: To determine the repeatability of this indirect method of assessment, bronchial challenge was performed with methacholine in nine children with stable asthma (age range 6-12 years) and was repeated 24 hours later. The response was determined by the fall both in forced expiratory volume in one second (FEV1) and in PtcO2. A modified tidal inhalation protocol was used in which quadrupling concentrations of methacholine were given, thereby reducing the time taken for the full challenge by almost half. The concentrations of methacholine that provoked a 20% decrease in FEV1 (PC20FEV1) and 15% and 10% falls in PtcO2 (PC15PtcO2, PC10PtcO2) were calculated. RESULTS: Repeatability measures, assessed as the 95% range for a single determination, were +/- 0.96 and +/- 1.12 doubling concentration differences respectively for PC15PtcO2 and PC10PtcO2 and +/- 0.80 for PC20FEV1. CONCLUSION: This challenge method using quadrupling concentrations and an indirect assessment of the response by PtcO2 was sufficiently repeatable for clinical use and compared favourably with repeated challenge assessed by FEV1. The PtcO2 method is simple and effort independent, and should prove particularly useful for measuring bronchial reactivity in young children.

Atopy, bronchial responsiveness, and symptoms in wheezy 3 year olds.

Fifty children with at least one hospital admission for acute lower airway obstruction in the first 2.5 years of life were assessed at 3 years of age to determine the relationship between atopy, bronchial responsiveness, and the pattern of their symptoms. Bronchial responsiveness was measured by assessing the effect of inhaled metacholine, using the change in transcutaneous oxygen tension (PtCO2) as an indirect measure of response. Symptom patterns were defined by the number of wheezing episodes associated with colds and the presence or absence of cough or wheeze unrelated to viral infections. Forty per cent of the children were found to be atopic by skin prick test or history. In contrast to the situation found in older children and adults, the non-atopic children had significantly greater bronchial responsiveness (lower mean concentration of methacholine causing a 20% fall in PtCO2, the PC20) than the atopic children and significantly more of them had an onset of respiratory symptoms in the first year of life. Cough and wheeze in the absence of colds was more frequently found in the atopic children as was the use of continuous medication. However, the number of reported acute episodes of wheeze associated with colds was the same in the two groups. The findings of the study suggest that in this hospital based group of children, acute wheeze associated with colds in the first three years of life is independent of the finding of atopy and that bronchial responsiveness in this age group may have a different pathogenesis from that in older subjects.

Skin prick testing using allergen-coated lancets: a comparison between a multiple lancet device and a single lancet applied with varying pressures.

Allergen-coated lancets have been developed to simplify skin prick testing. The effect of variation in application pressure on the response to prick tests was assessed in 20 atopic subjects, and the results compared to the response obtained with a newly devised multiple lancet device (MLD), capable of holding up to eight lancets and of providing a standardized application pressure. A positive weal (greater than or equal to 7 mm2) using light pressure occurred in 2/20 subjects, compared with 14/20 after moderate pressure. The largest weals were obtained using hard pressure and with the MLD (all 20 subjects obtained positive weals) and there was no significant difference in weal size between the two. There was evidence of a late reaction in 4/17 subjects with the MLD and with hard pressure applied to a single lancet, but in only one with moderate and in none with light pressure. Thus both the early and late skin responses are dependent on the pressure applied. The newly designed MLD allows skin prick tests to be performed using a standard pressure. It is also convenient for multiple allergen testing of uncooperative children.

Use of transcutaneous oxygen tension, arterial oxygen saturation, and respiratory resistance to assess the response to inhaled methacholine in asthmatic children and normal adults.

Respiratory resistance (Rrs6), transcutaneous oxygen tension (PtcO2), and oxygen saturation (SaO2) were measured during methacholine challenge in 15 asthmatic children and six normal adults. During bronchoconstriction, induced by a wide range of inhaled methacholine concentrations (0.5-256 g/l), the rise in Rrs6 was reflected by a fall in PtcO2 in all subjects. Although there was a significant mean fall in SaO2 at maximum bronchoconstriction there was no consistent relation between changes in SaO2 and Rrs6. The inhaled dose of methacholine causing a 40% increase in Rrs6 (PD40Rrs6) and a 20% fall in PtCO2 (PD20PtcO2) was calculated for each subject. There was no significant difference in mean PD40Rrs6 and PD20PtcO2, and the relation between the two was similar in the asthmatic children and the normal adults. It was therefore concluded that the measurement of PtcO2, but not SaO2, during methacholine challenge can be used for the assessment of bronchial responsiveness, and that it could prove particularly useful for children too young to cooperate with lung function tests.