E. Marek

Measurements of lactate in exhaled breath condensate at rest and after maximal exercise in young and healthy subjects

Arterial lactate concentrations, taken as indicators of physical fitness, in athletes as well as in patients with cardio-respiratory or metabolic diseases, are measured invasively from arterialized ear lobe blood. Currently developed micro enzyme detectors permit a non-invasive measurement of hypoxia-related metabolites such as lactate in exhaled breath condensate (EBC). The aim of our study is to prove whether this technology will replace the traditional measurement of lactate in arterialized blood. Therefore, we determined the functional relation between lactate release in EBC and lactate concentration in blood in young and healthy subjects at rest and after exhausting bicycle exercise. During resting conditions as well as after exhausting bicycle exercise, 100 L of exhaled air along with blood samples from the ear lobe was collected after stationary load conditions in 16 healthy subjects. EBC was obtained by cooling the expired air volume with an ECoScreen I (FILT GmbH, Berlin) condenser. The analysis was performed within 90 min using an ECoCheck ampere meter (FILT GmbH, Berlin). Lactate measurements were performed using a bi-enzyme sensor after lactate oxidase-induced oxidation of lactate to pyruvate and H2O2. The rates of lactate release via the exhaled air were calculated from the lactate concentration, the volume and the collection time of the EBC. The functional relation of lactate release in exhaled air and lactate concentration of arterial blood was computed. At rest, the mean lactate concentration in arterialized blood was 0.93 ± 0.30 mmol L(-1). At a resting ventilation of 11.5 ± 3.4 L min(-1), the collection time for 100 L of exhaled air, Ts, was 8.4 ± 2.9 min, and 1.68 ± 0.40 mL EBC was obtained. In EBC, the lactate concentration was 21.4 ± 7.7 µmol L(-1), and the rate of lactate release rate in collected EBC was 4.5 ± 1.7 nmol min(-1). After maximal exercise load (220 ± 20 W), the blood lactate concentration increased to 10.9 ± 1.8 mmol L(-1) and the ventilation increased to 111.6 ± 21.4 L min(-1). The EBC collection time decreased to 3.9 ± 1.9 min, and 1.20 ± 0.44 mL EBC were obtained in the recovery period after termination of exercise. The lactate concentration in EBC increased to 40.3 ± 23.0 µmol L(-1), and the lactate release in EBC increased to 13.6 ± 8.6 nmol min(-1) (p < 0.01). Assuming a volume of 4.3 mL water in 100 L of exhaled air (saturated with water at 37 °C), we calculated a lactate release at rest of 11.5 ± 4.3 nmol min(-1) and 48.6 ± 30.7 nmol min(-1) (p < 0.01) after exhausting exercise. Detectable releases of lactate in exhaled breath condensate were found already under resting conditions. During exhausting external load on a bicycle spiroergometer, an increase in the lactate concentration was found in arterialized blood along with an increased lactate release in EBC. The correlation between expiratory lactate release via EBC and lactate concentration in arterialized blood is studied in pursuing investigations.

L-Laktat und H2O2-Bestimmungen im Atemkondensat unter Ruhebedingungen sowie unter leichter bis mittelgradiger Belastung von jungen gesunden Probanden

BACKGROUND The recently developed microenzyme detectors make a non-invasive measurement of inflammatory markers and L-lactate in exhaled breath condensate (EBC) possible. In a group of young and healthy subjects, we examined whether L-lactate and H (2)O (2) can be detected in EBC. METHODS During resting conditions as well as at 60 and 120 Watt external load on a cycle ergospirometer 100 l exhaled air were collected under stationary load conditions from 19 healthy subjects. Exhaled breath condensate (EBC) was obtained by cooling the expired air volume. The analysis was performed within 90 min using an ECo-Check amperometer (Viasys Health Care). The H (2)O (2) measurement was performed amperometrically by means of a biosensor after chemical reaction catalysed by peroxidase. Lactate measurements were performed using a bienzyme sensor after lactate oxidase-induced oxidation of L-lactate to pyruvate and H (2)O (2). The rates of release of L-lactate in nmol/min und H (2)O (2) in pmol/min were calculated from the concentrations of L-lactate and H (2)O (2) in the EBC and the time of collection. RESULTS At rest 100 l exhaled air were collected in 10.6 +/- 5.1 min, and 0.99 +/- 0.3 ml EBC were obtained, at the 60 Watt step 1.23 +/- 0.47 ml EBC were collected in 6.7 +/- 1.8 min, and at 120 Watt 1.09 +/- 0.38 ml EBC in 4.8 +/- 0.8 min. At rest, there was a mean rate of L-lactate release of 3.3 +/- 3.1 nmol/min, which increased at the 60 Watt step to 8.4 +/- 5.1 nmol/min (p < 0.05), and at 120 Watt to 5.0 +/- 12.6 nmol/min (p < 0.02). The rate of L-lactate was proportional to the metabolic rate (r = 0.99). The rate of H (2)O (2) release at rest was 49.1 +/- 37.9 pmol/min, it increased at 60 Watt to 159 +/- 113 pmol/min (p < 0.05) and decreased at 120 Watt to 96.5 +/- 49.5 pmol/min (p < 0.05). CONCLUSIONS Significant measurable concentrations of L-lactate and H (2)O (2) in the exhaled breath condensate were found already under resting conditions. During external load, an increase in the L-lactate concentration was found, correlating with the metabolic rate. H (2)O (2) is an inflammatory marker, its concentration in the EBC was markedly increased during the first step of applied external load, but less during the second. A probable correlation between L-lactate concentration in EBC and arterialized blood will be studied in future investigations.

Exercise in Cold Air and Hydrogen Peroxide Release in Exhaled Breath Condensate

Athletes have changes in the lung epithelial cells caused by inhalation of cold and dry air. The exhaled breath condensate contains a number of mediators from the respiratory system and H(2)O(2) is described as a marker of airways inflammation. The aim of this study was to determine the influence of exercise combined with cold air on the H(2)O(2) release in the exhaled breath. Twelve males (23.1 ± 1.5 years) were randomly assigned at 2 different days (1 day rest) to perform a 50 min run (75-80% of their max. heart rate) under normal (N) laboratory (18.1 ± 1.1°C) or cold (C) field condition (-15.2 ± 3.1°C). Before and immediately after each run, the EBC was collected under laboratory conditions and was analyzed amperometrically. Prior to the two runs, H(2)O(2) concentrations were 145.0 ± 31.0 (N) and 160.0 ± 49.1 nmol/L (C) and theoretical release was 70.3 ± 37.1 (N) and 82.6 ± 27.1 pmol/min (C) (p > 0.05). After each run, H(2)O(2) concentration increased significantly to 388.0 ± 22.8 nmol/L (N) and 622.1 ± 44.2 nmol/L (C) (p < 0.05), along with an increase in the theoretical release: 249.2 ± 35.7 pmol/min (N) and 400.9 ± 35.7 pmol/min (C) (p < 0.05). We conclude that release of H(2)O(2) into the EBC takes place under both resting conditions and after exercise. The concentration and release of H(2)O(2) increased after exercise in cold air compared to resting and laboratory conditions, which points to an increase in inflammatory and oxidative stress.

Lungenfunktion im Alter: Brauchen wir neue Referenzwerte?

INTRODUCTION The structure of our aging population has significantly changed in the last three decades as have also the anthropometric data. Therefore, the question arises as to whether or not the largely accepted reference values for ventilatory lung function, which were suggested by the European Community for Coal and Steel (EGKS), may still be used today, since these values were obtained in the 1960s from subjects in a limited age range. For the elderly, the measured values are deduced by extrapolation beyond the range of reference equations which had been obtained in a different population. Therefore decisions concerning elderly and smaller subjects concerning remuneration due to impaired lung function after industrial exposure on the basis of EGKS values are questionable. METHODS We have examined lung function using pneumotachography for recording static lung volumes and flow-volume curves in 176 asymptomatic non-smoking males, aged 20 to 90 years, and correlated the results to the reference values of the EGKS, SAPALDIA and LuftiBus. RESULTS The age dependence of respiratory parameters (VC, FVC, FEV (1), FEV (1) %FVC, PEF, MEF (75,50,25)) for the healthy subjects can be described with a linear function (y = - m x age + n). The forced expiratory volume in one second, FEV (1), is calculated by FEV (1) = - 0.046 x age + 6.11; r = 0.88. Mean FEV1 for younger subjects was found to be 108 +/- 9.9 % of the EGKS reference values, 105 +/- 13.7 % in the middle-aged group and 97.3 +/- 12.4 % in the older subjects. All measured parameters concerning lung function can be described as linear functions of age which are steeper than those described by the EGKS reference values. The steeper slope in age dependency was also seen in other investigated parameters. The correlation of lung function parameters to height largely follows the EGKS predictions. CONCLUSIONS Measured lung function values of healthy younger and elderly subjects showed a close correlation to the extrapolated reference values of the EGKS. Our results relating to normal lung function justify an extrapolation of the reference equations beyond the common ranges of age while applying the same limitations as described for subjects in the middle-age range. Our results permit an extrapolation of EGKS values beyond the range of the reference values and can be used for the classification of impaired lung function in older subjects. The alternatively discussed reference equations of the SALPADIA Study, of NHANES and partially of the LuftiBus Study are higher, but do not cover all the necessary parameters and/or age ranges. A multicentric study for contemporary reference values should be performed in order to solve the problems concerning valid reference values.

Lung function in our aging population

Aims of investigationThe chronological age of the Caucasian population and their anthropometrical data have significantly changed within the last five decades. Therefore the question arises whether or not the commonly used reference values of the European Community (ECCS) for lung function may still be accepted today. Since these values were obtained in the 1960s from subjects in a limited age range. For the elderly, the measured values are deduced by extrapolation beyond the range of reference equations which had been obtained in a different population. Therefore decisions concerning elderly and smaller subjects concerning remuneration due to impaired lung function after industrial exposure on the basis of EGKS values are questionable.MethodsLung function tests were performed by pneumotachography, recording static lung volumes and flow-volume-curves in 262 asymptomatic non smoking males, aged 20 to 90 years. Measurements were performed with the MasterLab, or Pneumo-Screen systems (CareFusion, Höchberg). Results were compared to the reference values of ECCS, SAPALDIA and LuftiBus.ResultsFor simplicity analysis of age and height dependence of investigated respiratory parameters (VC, FVC, FEV1, FEV1%FVC, PEF, MEF75,50,25) can be described by linear functions (y = a * height - b * age + c). The forced expiratory vital capacity, FVC, was calculated by FVC = 0.0615*H - 0.0308*A - 4.673; r = 0.78. Mean FVC for younger subjects was found to be 104.7 ± 10.7% of the ECCS reference values and 96.5 ± 11.8% in older subjects. For most parameters investigated linear regressions on age were steeper than described by the ECCS reference values. The regression of lung function to height largely follows the ECCS prescriptions.SummaryBochum lung function values of younger healthy subjects were higher compared to the reference values of the ECCS and showed a steeper age descent. The alternatively discussed reference values of the SAPALDIA-, or LuftiBus-Study are higher, but do not cover all necessary parameters and/or the age range. A multi centre study for contemporary reference values is recommended.

Serial measurements of exhaled nitric oxide at work and at home: a new tool for the diagnosis of occupational asthma.

Whereas serial measurements of lung function at work and at home are a well-known diagnostic tool for the diagnosis of occupational asthma (OA), little is known about the serial measurements of non-invasive parameters such as exhaled nitric oxide (eNO). A 51-year-old baker with variable shortness of breath without relation to work was examined for suspected OA. Skin prick test showed weak sensitizations to wheat and rye flour (without sensitizations to environmental allergens) that were corroborated by in vitro testing (CAP class 3). Baseline FEV1 of 58% predicted and a decrease of forced expiratory volume in 1 s (FEV1) after placebo (sugar powder) of 17% did not allow inhalational challenge testing. The patient performed daily measurements of FEV1 and eNO for about a month during a holiday at home and at work. Whereas symptoms and FEV1 did not show differences between holidays and work periods, eNO showed a clear increase from below 10 ppb to a maximum of 75 ppb. A diagnosis of bakers asthma was made, and the patient quit his job immediately after medical advice. A year afterwards, the patient was still taking asthma medication, but his symptoms had improved, FEV1 had increased to 73% predicted, and eNO was 25 ppb. We conclude that serial measurements of eNO at home and at work may be a useful tool for the diagnosis of OA.

Lung Function at Age 18–25 Years: A Comparison of Different Reference Value Systems

The anthropometrical data of the Caucasian population have significantly changed within the last five decades. The European Community for Coal and Steel (ECCS) assumes a plateau phase and recommends the entry of 25 years old for calculation of reference values in this age range. The question arises if the commonly used reference recommendations for lung function of the ECCS can still be accepted. In the present study standardized spirometric lung function tests were performed by pneumotachography, recording lung volumes and flows (MasterScreen Pneumo, CareFusion, Höchberg) in asymptomatic nonsmoking subjects (202 females, 201 males), aged between 18 and 26, according to the ATS/ERS criteria. The results were compared with the reference recommendations of ECCS, SAPALDIA, LuftiBus, and Bochum (only males). All absolute lung function values showed a correlation (p< 0.05) with height. With respect to FVC and FEV(1), SAPALDIA and Bochum reference values were comparable and close to a 100 (range 97.6-101.4) %pred, whereas both ECCS and LuftiBus showed higher values (range 103.6-109.9%pred). The FEV(1)/FVC ratio was close to a 100 (range 97.6-101.7) %pred in all reference systems, whereas flows showed a wide variability between the reference systems (77.1-114.6%pred), single flows (e.g., 96.9-114.2%pred for MEF(50)) and males/females (males: 93.6-114.6%pred; females: 77.1-107.9%pred). We conclude that SAPALDIA reference values for FVC and FEV(1) should be used, as they better represent lung function in the age group. ECCS and LuftiBus reference values are appreciably (4-10%) lower. Differences between reference systems were less important for the FEV(1)/FVC ratio and lung flows.

Zur Reproduzierbarkeit der Wegstrecken beim 6-Minuten-Gehtest im Rahmen eines stationären Rehabilitationsaufenthaltes

UNLABELLED AIMS OF THE INVESTIGATION: The repetition of the 6-minutes walk test (6 MWT) in older patients is frequently performed in order to document the maximal walking distance, although it is not recommended in any guidelines on exercise tests and although there is common consent to save clinical resources in terms of time and staff. Therefore, we have examined whether and to what extent the repetition of the walk tests helps patients to get more familiar with this kind of exercise test. Thus the acquired physiological data should reliably describe the physical fitness of the patients at the beginning and at the end of their clinical rehabilitation. METHODS 35 patients performed their walk tests before and after 3 - 4 weeks of clinical rehabilitation. Each test has been repeated after one hour of recovery. The patients were instructed to walk during 6 minutes as fast as possible. They were equipped with a mobile pulse oximeter for recording oxygen saturation and heart rate. The distance, S, and the heart rate, fc, were measured. Measurements were performed every 30 seconds and recorded. The efficiency, E (E = S/6/fc), was calculated as the ratio of distance per minute and the mean heart rate during the test. RESULTS In the first test the patients walked 416 +/- 63 m at a heart rate of 104.7 +/- 15.7 beats/min, in the first repeated test 454 +/- 71 m at a heart of 106.3 +/- 17.4 beats/min. In the second test, after clinical therapy, they walked 438 +/- 58 m at a heart rate of 106.3 +/- 17.4 beats/min, in the second repeated test 473 +/- 56 m at 108.6 +/- 13.2/min. The difference of the walking distances of the tests at the entrance were found to be 38.4 +/- 26.2 m (+ 9.3 +/- 6.2%), at the end of clinical rehabilitation 35 +/- 26 m (+ 8.4 +/- 6.4%). Both differences are found to be independent from the distance of the first test. They are not significantly different. The efficiency was not significantly different in the initial and final test (0.673 +/- 0.129 and 0.689 +/- 0.085 m/beat, respectively). The difference in efficiency, when repeating the tests at the beginning, was: 0.053 +/- 0.062 m/beat; at the end of the rehabilitation: 0.042 +/- 0.047 m/beat. They are found to be similar. CONCLUSIONS The distances the patients walked in the repeated tests at the entrance and at the end of their clinical rehabilitation were, besides the calculated efficiency, E, significantly increased. However, the increases in distance and efficiency are identical on both occasions, therefore the repetition delivers no further information. The test should be performed without repetitions in clinical routine investigations. The patients performance in the second walk test with an unchanged distance at a lower heart rate reveals an improved physical fitness. This is solely described by an increase of efficiency, E. Therefore the introduction of E is a suitable measure of the quantified effect of exercise training, even if the patient is not cooperative during the tests. E is proved to be a suitable estimation for the assessment of physical fitness as a benefit of clinical rehabilitation.

Ein numerisches Verfahren zur Objektivierung der körperlichen Leistungsfähigkeit im Rahmen eines stationären Rehabilitationsaufenthaltes mittels 6-Minuten-Gehtest

UNLABELLED AIMS OF THE INVESTIGATION: The 6-minute-walk-test (6-MW) is an effective tool for measuring physical fitness in elderly patients. The increased walking distance is taken as a parameter for improved physical conditions. Frequently an unaltered walking distance is found after clinical treatment, but heart rate is significantly lower in the second challenge, indicating an improved physical fitness. This positive effect is not recognised when only the walking distance is analysed. METHODS An analysis of the 6-MW test was performed on 263 patients before and after 3 - 4 weeks clinical rehabilitation. In a control group of 26 patients 6-MW was repeated after recovery at the beginning and the end of the clinical treatment. Instrumented by a mobile pulse oximeter for recording oxygen saturation and heart rate, patients were instructed to walk as fast as they can do during 6 minutes. Measurements were performed every 30 seconds and printed out. Two new parameters, efficiency (E = S/f (C)), the ratio of distance and mean heart rate, and the theoretical increase in walking distance (S (z) = Delta f (C1)/Delta f (C2) x S (2) - S (1)) were introduced and tested. S (z) = theoretical increase in distance, Delta f (C1) = difference in heart rate at rest and mean heart rate at steady state during the first walk test with distance, S1. Delta f (C2), and S2 are measured during the second walk. Thus, the increase in distance is calculated under the assumption that the second walk test would have been performed by the patient with the same difference in heart rate that he/she achieved in the first walk. RESULTS The patient groups walked 353 +/- 80 m at 106 +/- 14.3 beats/min in the 1st. 6-MW and 368 +/- 76.9 m at a heart rate of 105 +/- 14.0 beats/min in the final test. The increase of the walking distance was most significant in patients with shorter distances in the 1st 6-MW. A significant increase in the walking distance and in efficiency was found in patients with shorter walking distances or lower heart rates in the final test, using the numerical procedure described above. CONCLUSIONS The patients performance of the second walk test with an unchanged distance at a lower heart rate reveals an improved physical fitness. This is solely described by an increase by the parameter of efficiency, E. The calculation of the parameter, Sz, theoretical difference in walking distance (i. e., theoretical increase in almost all tests) provides a quantification of the effect of exercise training, even if the patient is not cooperative during the tests. Both parameters have proved to be suitable estimations for the assessment of physical fitness as a beneficial effect of clinical rehabilitation.