Márk Kollai

Exhaled biomarkers in lung cancer

Lung cancer is the leading cause of cancer death. Results of therapeutic interventions are particularly discouraging when the disease is discovered in an advanced stage. Early diagnosis is limited by the fact that the disease usually develops asymptomatically and available screening methods do not fulfil the requirements for reliable discrimination between patients with lung cancer and subjects not suffering from the disease. Breath sampling is completely noninvasive and provides a potentially useful approach to screening lung cancer. Exhaled biomarkers contain both volatile and nonvolatile molecules. The profile of volatile organic compounds is different in patients with lung cancer than in control subjects. In exhaled breath condensate, the proteomic profile of breath from cancer patients differs from that of healthy smokers. We reviewed the scientific evidence demonstrating that a unique chemical signature can be detected in the breath of patients with lung cancer and that the exhaled breath biomarker profile could aid clinical decision making.

Maturation of Cardiovagal Autonomic Function From Childhood to Young Adult Age

Background—Cardiovagal autonomic control declines with age in adult subjects, which is related in part to increasing stiffness of the barosensory vessel wall. It is not known, however, whether autonomic function changes with age in children. Methods and Results—We studied 137 healthy subjects divided into 4 age groups: group 1, 7 to 14 years; group 2, 11 to 14 years; group 3, 15 to 18 years; and group 4, 19 to 22 years. Brachial artery pressure was measured by sphygmomanometry and continuous radial artery pressure and carotid artery pulse pressure (&Dgr;P) by applanation tonometry. The R-R interval was derived from the ECG. Autonomic function was assessed by spontaneous sequence and frequency-domain indices, which indicate the extent of coupling between fluctuations in heart rate and systolic pressure. Carotid artery diastolic diameter (DD) and pulsatile distension (&Dgr;D) were measured by echo wall tracking; carotid compliance coefficient (CC) was defined as &Dgr;D/&Dgr;P and distensibility coefficient as 2&Dgr;D/DD · &Dgr;P. From group 1 to group 3, spontaneous indices increased significantly (18.1±1.7 versus 33.3±4.0; 14.4±1.1 versus 25.5±22; 12.9±1.1 versus 20.8±2.0; and 6.4±0.6 versus 16.2±1.4 ms/mm Hg [mean±SEM] for Seq+, Seq−, LF&agr;, and LFgain, respectively), with no significant changes afterward. CC and DC were inversely proportional to age (r=−0.49 and −0.62, respectively, P<0.001). The efficiency of neural integrative mechanisms, estimated as the ratio of spontaneous indices and CC, more than doubled from group 1 to group 3. Spontaneous indices were linearly related to measures of cardiac vagal activity. Conclusions—The increase in spontaneous indices from early childhood to adolescence, despite gradual stiffening of the carotid artery, may indicate improved cardiovagal autonomic function, which is most likely a result of maturation of neural mechanisms, attaining peak level at adolescence.

Exhaled breath condensate pH standardised for CO2 partial pressure

Exhaled breath condensate (EBC) pH is considered to reflect the acid-base balance of the airways. Current pH measurements do not take into account the effect of CO2. The aim of the present study was to determine the effect of condensate CO2 partial pressure on pH and to provide a more precise mode of EBC pH determination. Condensate pH and CO2 partial pressure were measured in parallel from 12 healthy volunteers and 12 asthmatics using a blood gas analyser in neat, argon de-aerated and CO2-loaded samples. The regression analysis was used to test the relationship between pH and CO2, and to calculate the pH at a CO2 level of 5.33 kPa (physiological alveolar CO2 partial pressure). Reproducibility of different pH readings was compared using the Bland–Altman test. Condensate CO2 concentration was variable both in neat and argon de-aerated samples. There was a close negative logarithmic relationship between CO2 and pH. Calculation of pH at a CO2 level of 5.33 kPa provided reproducibility approximately six times as good as that of the currently used measurements. Condensate CO2 partial pressure influences pH measurements. Determination of pH at a standard CO2 level provides the most reproducible condensate pH values to date.

Adenosine level in exhaled breath increases during exercise-induced bronchoconstriction

In asthmatic patients, airway obstruction provoked by exercise challenge is accompanied by an increase in plasma adenosine level. In this study, the current authors investigated if exercise-induced bronchoconstriction was associated with local changes of adenosine concentration in the airways. Oral exhaled breath condensate (EBC) collection (5-min duration) and forced expiratory volume in one second (FEV1) measurements were performed at rest (baseline) and 4–8 times after treadmill exercise challenge in healthy and asthmatic subjects. Adenosine concentration in EBC was determined by HPLC. Observations indicated that physical exercise results in bronchoconstriction together with a significant increase of adenosine level in EBC in asthmatic patients (mean±sd maximal fall in FEV1 27±13%; associated increase in adenosine 110±76% as compared to baseline), but not in healthy control subjects. Exercise-induced changes in adenosine concentration correlated significantly with the fall in FEV1 values in asthmatic patients. In conclusion, the observed increase in adenosine concentration of oral exhaled breath condensate most probably reflects changes in the airways during exercise-induced bronchoconstriction. Due to its known bronchoconstrictor property in asthma, adenosine may contribute to the development of bronchospasm.

Heart rate variability after complete autonomic blockade in man

In a number of studies, using the autoregressive model for frequency domain analysis of R-R interval fluctuations, the low frequency (LF) component (centered at about 0.1 Hz) is claimed to index sympathetic activity level. The aim of this study was to investigate the mediation mechanism of the LF component by pharmacological blockade. Our results support earlier findings, obtained with the use of fast Fourier transformation, that in supine subjects spectral components of R-R interval variability at around 0.1 Hz are mediated mainly by cholinergic mechanisms. Therefore, the use of the LF component as sympathetic index appears questionable.

Increase of carotid artery stiffness and decrease of baroreflex sensitivity in exfoliation syndrome and glaucoma

Aim: To investigate the distensibility of the common carotid artery (CCA), baroreflex sensitivity (BRS) and its relation to plasma homocysteine concentration in exfoliation syndrome or exfoliation glaucoma (XFS/XFG). Methods: Homocysteine concentrations were measured in 30 XFS/XFG patients and 18 age matched controls. In 21 patients and 17 controls the end diastolic diameter of the CCA and pulsatile distension were measured and BRS was calculated. Results: There was no significant difference between the groups in sex distribution, age, heart rate, blood pressure, systemic diseases, or medication. In XFS/XFG patients homocysteine concentration was significantly elevated (unpaired t test, p = 0.023), and CCA stiffness was higher (p<0.05), while strain, cross sectional compliance coefficient, distensibility, and BRS were significantly reduced compared to the controls (Mann-Whitney U test, p⩽0.013 for each parameter). In XFS/XFG patients a positive correlation was found between age and plasma homocysteine level (Pearson’s correlation, r = 0.490, p = 0.007), and a negative correlation between age and BRS (Kendall’s correlation r = −0.374, p = 0.021), as well as between homocysteine concentration and BRS (Kendall’s correlation r = −0.377, p = 0.024). No correlation was seen between these variables in the control group. Conclusions: These results suggest a pathological large artery function as well as altered parasympathetic vascular control in XFS/XFG which increases with age and with higher homocysteine concentration.

Lack of mitochondrial nitric oxide production in the mouse brain

Based on our initial finding that the nitric oxide (NO) sensitive fluorochrome diaminofluorescein (DAF) was localized to mitochondria in cultured primary neurons, we investigated whether brain mitochondria produce NO through a mitochondrial NO synthase (mtNOS) enzyme. Isolated brain mitochondria were loaded with DAF and subjected to flow cytometry analysis. Neither the application of NOS inhibitors nor the genetic disruption of either NOS gene diminished the DAF‐fluorescence. However, peroxynitrite scavengers reduced the mitochondrial DAF fluorescence, indicating that the DAF signal is not specific to NO. Chemiluminescence detection in the head space gas and a Clark‐type NO‐sensitive electrode in the solution failed to detect NO release in brain mitochondria. NOS activity in mitochondria was only 1% of the whole brain NOS activity level, which may be attributed to extramitochondrial contamination. Extensive immunoblotting and immunoprecipitation experiments failed to show the presence of endothelial, neuronal, or inducible NOS in mouse brain mitochondria using a variety of primary antibodies. Arginine, calmodulin or 2,5‐ADP affinity purification protocols successfully concentrated eNOS and nNOS from full brain tissue but failed to show any signal in mitochondria. We conclude that mouse brain mitochondria do not contain NOS isoforms, nor do they produce NO through a NOS‐dependent mechanism.

Mitochondria produce reactive nitrogen species via an arginine-independent pathway

We measured the contribution of mitochondrial nitric oxide synthase (mtNOS) and respiratory chain enzymes to reactive nitrogen species (RNS) production. Diaminofluorescein (DAF) was applied for the assessment of RNS production in isolated mouse brain, heart and liver mitochondria and also in a cultured neuroblastoma cell line by confocal microscopy and flow cytometry. Mitochondria produced RNS, which was inhibited by catalysts of peroxynitrite decomposition but not by nitric oxide (NO) synthase inhibitors. Disrupting the organelles or withdrawing respiratory substrates markedly reduced RNS production. Inhibition of complex I abolished the DAF signal, which was restored by complex II substrates. Inhibition of the respiratory complexes downstream from the ubiquinone/ubiquinol cycle or dissipating the proton gradient had no effect on DAF fluorescence. We conclude that mitochondria from brain, heart and liver are capable of significant RNS production via the respiratory chain rather than through an arginine-dependent mtNOS.

Multiple modes of operation of cardiac autonomic control: development of the ideas from Cannon and Brooks to the present

Since Cannons time much emphasis has been placed on reciprocal control of the organ function by the autonomic nerves while other modes of control has been neglected. In our laboratory, prompted by Dr. Brooks, we initiated the study of the autonomic control of the heart by recording simultaneously the activity of cardiac sympathetic and parasympathetic nerves under a variety of conditions in chloralose anesthetized dogs. We have demonstrated that the central nervous system exercises reciprocal as well as non-reciprocal pattern of control over the two arms of the autonomic outflows under different physiological and behavioral conditions, and that each mode of control has an important functional significance.

Static and dynamic changes in carotid artery diameter in humans during and after strenuous exercise

Arterial baroreflex function is altered by dynamic exercise, but it is not clear to what extent baroreflex changes are due to altered transduction of pressure into deformation of the barosensory vessel wall. In this study we measured changes in mean common carotid artery diameter and the pulsatile pressure: diameter ratio (PDR) during and after dynamic exercise. Ten young, healthy subjects performed a graded exercise protocol to exhaustion on a bicycle ergometer. Carotid dimensions were measured with an ultrasound wall‐tracking system; central arterial pressure was measured with the use of radial tonometry and the generalized transfer function; baroreflex sensitivity (BRS) was assessed in the post‐exercise period by spectral analysis and the sequence method. Data are given as means ±s.e.m. Mean carotid artery diameter increased during exercise as compared with control levels, but carotid distension amplitude did not change. PDR was reduced from 27.3 ± 2.7 to 13.7 ± 1.0 μm mmHg−1. Immediately after stopping exercise, the carotid artery constricted and PDR remained reduced. At 60 min post‐exercise, the carotid artery dilated and the PDR increased above control levels (33.9 ± 1.4 μm mmHg−1). The post‐exercise changes in PDR were closely paralleled by those in BRS (0.74 ≤r≤ 0.83, P < 0.05). These changes in mean carotid diameter and PDR suggest that the mean baroreceptor activity level increases during exercise, with reduced dynamic sensitivity; at the end of exercise baroreceptors are suddenly unloaded, then at 1 h post‐exercise, baroreceptor activity increases again with increasing dynamic sensitivity. The close correlation between PDR and BRS observed at post‐exercise underlies the significance of mechanical factors in arterial baroreflex control.