Effects of genetic vs. environmental factors on cardiovascular autonomic function: a twin study.

AIMS: Cardiovascular autonomic function is often assessed in patients with diabetes by measuring heart rate variability and baroreflex sensitivity, the heritability of which is not fully understood. The present study was aimed to determine the effects of genetic and environmental factors on heart rate variability and baroreflex sensitivity in monozygotic and dizygotic adult healthy twin pairs. METHODS: A total of 101 (63 monozygotic, 38 dizygotic) adult twin pairs (n = 202; mean age 44.3 years) were investigated. Anthropometric variables and serum metabolic markers were measured, while environmental characteristics were evaluated by questionnaires. Linear and spectral indices of heart rate variability and baroreflex sensitivity were determined by non-invasive methods. All measurements were adjusted for age and gender (model 1) and for all significantly relevant covariates (model 2). Heritability A-C-E structural equation models were used for characterizing the proportion of additive genetic, shared and unshared environmental influences. RESULTS: Genetic influence of different cardiovascular autonomic indices was estimated between 10.3 and 39.4%, common environmental influence was found between 0.0 and 33.2%, while unshared environmental influence was observed between 60.6 and 81.4% in model 1 analysis. In multivariable-adjusted heritability estimates (model 2), the magnitude of the genetic effects decreased to 0.0%, common environmental influence was nearly unchanged (values between 4.4 and 14.5%), while unshared environmental influence slightly increased (values between 85.5 and 96.5%). CONCLUSIONS: Unshared environmental but not genetic factors have substantial influence on cardiovascular autonomic function, suggesting that appropriate treatment of all modifiable environmental factors is of importance in order to prevent or ameliorate cardiovascular autonomic neuropathy.

Nitrative stress and poly(ADP-ribose) polymerase activation in healthy and gestational diabetic pregnancies.

AIMS/HYPOTHESIS: Increased oxidative-nitrosative stress, poly(ADP-ribose) polymerase (PARP) activation and subsequent cellular damage play important roles in the complications of both diabetes mellitus and pregnancy. Our aim was to investigate nitrative stress and PARP activity levels during normal and gestational diabetic (GDM) pregnancy in both maternal and fetal tissues. METHODS: Blood samples were collected during pregnancy (weeks 16-29 and 36-40), and placental and umbilical cord tissues were harvested after delivery from healthy volunteers and GDM patients subjected to a carbohydrate-restricted diet or insulin treatment. Immunohistochemical staining was performed on leucocytes and tissue sections using anti-nitrotyrosine (NT), anti-poly(ADP-ribose) (PAR) and anti-apoptosis inducing factor antibodies. RESULTS: In healthy pregnancies the intensity of NT and PAR staining of leucocytes correlated positively with gestational week (R (2) = 0.43, p < 0.01 and R (2) = 0.49, p < 0.001, respectively). In patients on a carbohydrate-restricted diet PAR staining was already strong in weeks 16-29 (p < 0.001 vs control) and did not increase further. In weeks 16-29 there was a correlation between PAR staining and the 2 h value of the oral glucose tolerance test (R (2) = 0.49, p < 0.001). Patients with the highest level of leucocyte PARP activity later required insulin therapy, which decreased the intensity of NT and PAR staining. Placental and umbilical cord tissues also had a higher level of nitrative stress markers in GDM pregnancies, but the highest level of PARP activity was observed after insulin therapy. CONCLUSIONS/INTERPRETATION: Continuous elevation of tyrosine nitration and PARP activation may be considered physiological during pregnancy. However, the high level of PARP activity in early pregnancy may signal the subsequent development of severe GDM.

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.

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< or =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.

Relation between bronchial and cardiac vagal tone in healthy humans.

A generally increased vagal nervous activity was suggested to exist in bronchial asthma; however, this contention has never been fully tested. Before studying vagal activity in asthma, we raised the question if there existed any relation between vagal outflows to different target organs in the physiologic state. The specific aim of this study was to determine the relation between the resting level of bronchial and cardiac vagal activity in healthy volunteers. Bronchial and cardiac vagal tone were measured in 12 subjects as the change of airway resistance (Raw) and heart period (HP) in response to complete cholinergic blockade. It was found that after intravenous atropine administration, Raw was reduced from 1.30 +/- 0.41 cm H2O/L/s to 0.51 +/- 0.22 cm H2O/L/s and HP was reduced from 789 +/- 84 ms to 506 +/- 41 ms. All the values are given as mean +/- SD. Both the bronchial and cardiac responses exhibited considerable interindividual variability, but there was no significant correlation between them (R = 0.22, p = 0.50). It was concluded that under resting conditions, vagal control of bronchial tone and HP were not related in healthy human subjects.

Study of cardiac sympathetic and vagal efferent activity during reflex responses produced by stretch of the atria.

(1) In chloralose anesthetized dogs, effects of the left and the right atrial stretch were studied in the same animal. Stretch of the sino-atrial region of the right atrium produced acceleration of the heart rate during, and reversal of response at the termination of, the stretch. Stretch of the left pulmonary vein-atrial junctional region evoked an initial decrease followed by an increase in heart rate. The responses were similar in all animals, despite initial heart rates ranging from 120 to 200 beats per minute. (2) Activity in vagal and sympathetic nerve branches innervating the heart was recorded simultaneously. Care was taken to identify the vagal fibers innervating the heart, and record their activity without contamination of sympathetic impulses. The right atrial stretch evoked an augmentation of sympathetic activity which reached its peak at 20 sec after the beginning of stimulus. The stimulus slightly increased the vagal activity; this change occurred slowly and reached its peak in about 40-60 sec after stretch. At the release of stretch, sympathetic activity generally showed a reversal of response, i.e. activity was inhibited for 30 sec. (3) Stretch of the left atrium produced biphasic changes in cardiac sympathetic nerves; their activity was strongly inhibited for the first 15 sec, then augmented throughout the remainder of the stretch. This effect lasted 30 sec after the cessation of stimulus. Effects on vagal cardiac nerve fibers were smaller; mild augmentation in activity was produced. The onset of this effect was faster than that seen in case of the right atrium stretch. (4) Reciprocal action between vagal and sympathetic cardiac nerves was obvious only in the early phase of left atrium stretch. Effects on the heart were determined by balances in activity of these antagonistic nerves. In contrast with what occurred in cardiac reflexes, carotid sinus distension even in the same animal produced a large increase in vagal activity, and near complete inhibition of sympathetic nerve activity. Thus, good reciprocal action between the two sets of nerves was demonstrated. A difference in the two types of reflexes was thus revealed. (5) Stretch of the right atrium evoked during carotid sinus distension caused an increase in heart rate from the new low level which was produced by baroreceptor activation. Vagal activity which was greatly augmented by sinus distension was decreased by atrial stretch, while previously inhibited sympathetic activity due to sinus distension was augmented by stretch of the atrium. The effect of stretch on vagal activity seems to depend to a degree on the prestimulus level. It is of interest that the powerful baroreceptor reflexes do not mask the cardiac reflexes studied. (6) The relative importance of sympathetic and vagal efferents to atrial stretch reflexes was discussed.

Reduced cardiac vagal excitability in hyperthyroidism.

A great deal of uncertainty persists regarding the exact nature of the interaction between autonomic nervous activity and thyroid hormones in the control of heart rate. In the present work we investigated whether reduced vagal influence could contribute to the tachycardia in hyperthyroidism. Vagal excitability was studied in ten hyperthyroid patients. Prolongation of R-R interval in response to carotid baroreceptor stimulation by neck suction was found to be less in the hyperthyroid state compared to the control state after therapy. The extent of nocturnal bradycardia and the vagal excitatory response to the central effect of low dose atropin was significantly reduced in hyperthyroid patients compared to euthyroid controls. We concluded that in the hyperthyroid state cardiac vagal motoneurones were in low excitability state, and speculated that the inhibition might have resulted from thyroid hormone action on CNS structures integrating autonomic function and behaviour.

Measurement of aortic arch distension wave with the echo-track technique.

The aim of this study was to use the echo-track method for measuring aortic arch diameter, distension waveform and elastic parameters. Data were obtained from 50 healthy volunteers of 32 +/- 15 y (mean +/- 1 SD). The aortic arch was interrogated from the suprasternal position with M-mode ultrasonography using a 3.5-MHz transducer; diameter and distension waves were determined by means of an echo-track algorithm (WTS, Pie Medical); arterial blood pressure was measured in the arm with sphygmomanometry. Aortic arch diameter, distension, distensibility and compliance were 24.55 +/- 2.99 mm, 2199 +/- 726 micrometer, 3.9 +/- 1.4. 10(-3) mmHg(-1) and 1.86 +/- 0.61 mm(2). mmHg(-1), respectively. Intrasession, interobserver and intersession variability was less than 10%, 10% and 18%, respectively. It is concluded that aortic arch distension wave can be recorded noninvasively with acceptable reproducibility, allowing assessment of aortic elastic parameters, and yielding insight into pressure wave reflection within the arterial system.

Comparison of aortic arch and carotid sinus distensibility in humans--relation to baroreflex sensitivity.

We compared aortic arch (AA) and carotid sinus (CS) distensibility, and determined the relationship between baroreflex sensitivity (BRS) and distensibility coefficients of AA and CS (DCAA and DCCS, respectively). In 47 healthy 19-71-year-old subjects, AA and CS end-diastolic diameter (D) and pulsatile distension (delta D) was measured with ultrasound and arterial pressure by sphygmomanometer and Finapres. DC was calculated as 2delta D/(D x delta P), where delta P is the pulse pressure. BRS was determined by the sequence method. Data are given as mean +/- S.D.; significance was set at p < 0.05. DCAA and DCCS were linearly related across subjects (r = 0.80, p < 0.001). No difference between DCAA and DCCS (3.8 +/- 1.4 x 10(-3) and 4.2 + 2.2 x 10(-3) mm Hg(-1), respectively) was found for all subjects (paired t-test). However, at ages < 35 years, DCCS exceeded DCAA, whereas this relation was reversed at ages > 35 years. Age accounted for 53% of variability in DCAA and 73% of variability in DCCS. BRS was related to DC (linear regression), with the BRS-DCAA relation being steeper and closer (r = 0.73, p < 0.001) than the BRS-DCCS relation (r = 0.54, p < 0.05). It is suggested that aortic baroreceptors may have a dominant role in heart rate control.

Preserved structural and functional characteristics of common carotid artery in properly treated normoglycemic women with gestational diabetes mellitus.

Women with gestational diabetes mellitus (GDM) are at high risk of subsequently developing type 2 diabetes mellitus which is an important cardiovascular risk factor. We have evaluated whether preclinical morphological and functional arterial changes are present in GDM. Diameter, intima-media thickness (IMT), intima-media cross-section area (IMCSA) and elasticity features (compliance, distensibility coefficient, circumferential strain, stiffness index (SI) α and ß, incremental elastic modulus) of the common carotid arteries (CCA) were studied in the 3rd trimester in 25 women with GDM, and 17 normal pregnant women matched for age and body mass index using an ultrasonographic vessel wall-movement tracking system and applanation tonometry. Mean IMT, IMCSA and SI α tended to be larger, whereas compliance was smaller in women with GDM but none of these differences were significant. Serum glucose (4.99 ± 0.51 vs. 4.79 ± 0.61 mmol/L, p=0.37) and HbA1c (5.33 ± 0.27 vs. 5.36 ± 0.47 mmol/L, p=0.85) proved normoglycemia in both groups. In conclusion, by the combination of methods we applied in this case control study, neither morphological nor functional characteristics of large elastic arteries differ significantly between well-treated normoglycemic women with GDM and non-diabetic pregnant women in the 3rd trimester.

Adenosine triphosphate in exhaled breath condensate of healthy subjects and patients with chronic obstructive pulmonary disease.

OBJECTIVES: The effect of hypoxic relapse of chronic obstructive pulmonary disease (COPD) on lung adenosine triphosphate (ATP) concentration was studied measuring ATP in exhaled breath condensate (EBC). SUBJECTS: Thirty COPD patients with severe exacerbation, thirteen healthy non-smokers and thirteen healthy smokers. METHODS: ATP was detected using a luciferin-luciferase assay, dilution of airway droplets in EBC was assessed measuring sample conductivity. RESULTS: ATP concentrations were similar in COPD patients, non-smoking and smoking healthy individuals (141 +/- 44, 115 +/- 21 and 90 +/- 15 pM; p = 0.66). After treatment oxygenation of COPD patients improved (6.85 +/- 1.29 kPa vs. 8.20 +/- 1.28 kPa, p < 0.001), but EBC ATP concentration was similar to that of admission (p = 0.84). There was no correlation between EBC ATP concentration and airway droplet dilution. CONCLUSION: ATP detected in EBC indicates the presence of ATP in airway lining fluid. Lack of difference in ATP concentration between health and COPD suggests that airway ATP level is under complex control of multiple factors.

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.

S-nitrosoglutathione-containing hydrogel increases dermal blood flow in streptozotocin-induced diabetic rats.

BACKGROUND: Endothelial dysfunction is characterized by decreased vasodilatory capacity of the arterioles mainly due to the reduced release of nitric oxide (NO). Application of NO donors may prevent or even reverse the consequences of endothelial dysfunction, such as diabetic leg ulcers. OBJECTIVES: To investigate the vasodilatory capacity and the possible side-effects of topical application of an NO donor-containing hydrogel in diabetic rats. METHODS: S-nitrosoglutathione (GSNO) was incorporated in Pluronic F127 hydrogel and applied on the foot sole skin of healthy and streptozotocin-induced diabetic rats. Blood flow was monitored using a laser-Doppler probe. Nitrotyrosine formation, a possible side-effect of GSNO action, was evaluated by Western blotting of skin protein extracts. Systemic circulatory side-effects were investigated by monitoring blood pressure and heart rate during the application. RESULTS: The hydrogel alone did not induce any changes in microvascular flow, while GSNO-containing hydrogel caused a twofold increase in perfusion. This effect was similar in diabetic and healthy animals. Topical GSNO application did not increase the nitrotyrosine content of skin proteins, nor did it have any effect on blood pressure or heart rate. CONCLUSIONS: Dermal application of GSNO may be an effective treatment for promoting the local vasodilation in both healthy and diabetic states, without inducing protein nitration or alterations in blood pressure or heart rate.

Determinants of baroreflex function in juvenile end-stage renal disease.

Arterial baroreflex sensitivity (BRS) is markedly reduced in middle-aged patients with end-stage renal disease (ESRD), due to the combined effects of aging, arterial stiffening, and autonomic neuropathy. Much less is known about the effects of ESRD on arterial baroreflex in juvenile patients. Therefore, we investigated baroreflex function and its relation to carotid artery elasticity and heart rate variability in children and young adults with ESRD. We studied 42 subjects (9-30 years): 14 patients on maintenance hemodialysis (HD), 14 renal transplant recipients (RT), and 14 healthy control subjects (C). Baroreflex function was determined by pharmacological (BRS) and spontaneous (sequence and spectral indices) techniques. Carotid artery elasticity was characterized by stiffness index beta. Heart rate variability was assessed using time and frequency domain measures. Data are expressed as mean+/-s.d. BRS was markedly reduced in HD as compared to C (10.0+/-4.2 vs 25.7+/-5.9 ms/mm Hg); spontaneous indices were reduced to similar extent. Carotid artery stiffness was approximately 50% higher in HD than in C and was inversely related to BRS. Heart rate variability was also compromised in HD, and was directly related to spontaneous indices. No significant differences existed in any of these variables between RT and C. Decreased baroreflex function in juvenile HD is partly due to loss of carotid artery elasticity and partly due to impaired heart rate variability. Renal transplantation may partly prevent impairment or improve compromised baroreflex function in young patients with ESRD.

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.

Responses in cutaneous vascular tone to transient hypoxia in man.

The study was undertaken to determine the changes in skin circulation in response to systemic hypoxia. Transient hypoxia was induced in 24 healthy subjects by breathing 8% O2 in N2 for 90 s. Arterial oxygen tension was measured transcutaneously (tcpO2), photoplethysmography was employed to monitor changes in cutaneous vessel tone, and respiratory tidal volumes were measured by a Fleisch pneumotachograph. End tidal CO2 concentration, ECG, heart rate, finger skin resistance, and finger skin temperature were monitored continuously as well. Multi-unit skin sympathetic activity was recorded in the median nerve at wrist level by using Tungsten microelectrodes. By the end of the 90 s hypoxic test period tcpO2 was reduced from the control value of 95 +/- 1.8 mm Hg to 51.7 +/- 2.8 mm Hg; respiratory tidal volume increased from the resting value of 585 +/- 22 ml to 746 +/- 38 ml, while photoplethysmographic pulse amplitude decreased to 60% of control, together with a 0.22 +/- 0.03 degree C drop in skin temperature. The values are +/- S.E. In half of the cases skin resistance was reduced as well. Skin sympathetic activity increased during hypoxia and each major burst was followed by a reduction in pulse amplitude. The respiratory and photoplethysmographic responses exhibited similar time courses, with corresponding peaks. Voluntary hyperventilation for 90 s with room air also produced an initial reduction in pulse amplitude; however, it recovered within 60 s. It is concluded that in conscious human subjects, transient systemic hypoxia leads to constriction of cutaneous vessels in the hand, and that the vasoconstriction is the result of increased traffic in sympathetic efferent fibers. Skin vasoconstriction can develop independently of respiratory changes; usually the concomitant hyperventilation facilitates the cutaneous response.

Cardiac vagal tone in generalised anxiety disorder.

The aim of the present study was twofold: firstly, to determine cardiac vagal tone in subjects with generalised anxiety disorder directly, using an invasive pharmacological method; and secondly, to test whether the non-invasive method of measuring the amplitude of respiratory sinus arrhythmia (RSA) gives a reliable estimate of cardiac vagal tone in this form of anxiety disorder. Comparison of baseline physiological measures of anxious and control subjects revealed that cardiac vagal tone and heart rate were not different in the two groups of subjects, whereas length of the respiratory cycle and amplitude of respiratory sinus arrhythmia were reduced in the former group. Because of the higher respiratory rates of anxious subjects, the RSA method was found to underestimate cardiac vagal tone in generalised anxiety disorder.

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

Since Cannon's 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.