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Departments of 1 Chemistry and Physics and 4 Biology, Asbury College, Wilmore 40390-1198; 2 Department of Physiology, University of Kentucky College of Medicine, Lexington 40536-0298; and 3 Center for Biomedical Engineering, University of Kentucky, Lexington, Kentucky 40506-0070
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ABSTRACT |
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This study explores the functional association between renal sympathetic nerve traffic (NT) and arterial blood pressure (BP) in the very-low-frequency range (i.e., <0.1 Hz). NT and BP (n = 6) or BP alone (n = 17) was recorded in unanesthetized rats (n = 6). Data were collected for 2-5 h, and wavelet transforms were calculated from data epochs of up to 1 h. From these transforms, we obtained probability distributions for fluctuation amplitudes over a range of time scales. We also computed the cross-wavelet power spectrum between NT and BP to detect the occurrence in time of large-amplitude transient events that may be important in the autonomic regulation of BP. Finally, we computed a time sequence of cross correlations between NT and BP to follow the relationship between NT and BP in time. We found that NT and BP follow comparable self-similar scaling relationships (i.e., NT and BP fluctuations exhibit a certain type of power law behavior). Scaling of this nature 1) points to underlying dynamics over a wide range of scales and 2) is related to large-amplitude events that contribute to the very-low-frequency variability of NT and BP. There is a strong correlation between NT and BP during many of these transient events. These strong correlations and the uniformity in scaling imply a functional connection between these two signals at frequencies where we previously found no connection using spectral coherence.
wavelet analysis; sympathetic nervous activity; cross correlation; blood pressure fluctuations; transient events; self-similar invariance
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INTRODUCTION |
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ARTERIAL BLOOD PRESSURE (BP) and sympathetic nerve traffic (NT) are well known to be functionally coupled, or highly coherent, to maintain normal homeostatic function. In the unanesthetized rat, for example, oscillations in sympathetic NT that repeat in ~2.5 s are clearly very tightly coupled with fluctuations in BP that have a similar period (3). This high coherence can be explained on the basis of the baroreflex (4). This 0.4-Hz rhythm appears to be a natural instability within this reflex, similar to a resonance in an electrical circuit or mechanical system produced by the time constants and delays in the constituent physical and neuronal phenomena. A similar phenomenon in the rabbit occurs at 0.3 Hz (13) and in the human at 0.1 Hz (7, 17). Conversely, the coherence between sympathetic activity and BP in the rat falls to very low values below a frequency of 0.1 Hz (3). In other words, the mathematical processes used to compute the coherence do not detect a close functional relationship between rhythmic changes in BP and corollary changes in sympathetic NT, or vice versa, that require more than ~10 s to repeat. A weak coupling between sympathetic activity and BP within the very low frequencies (i.e., <0.1 Hz) is unexpected, because the baroreflex is widely credited with minimizing BP fluctuations during, for example, postural changes that certainly fall at least within the upper limits of the low-frequency range.
Stationarity of a signal, or "time series," is required in spectral analysis, which is the basis of the coherence computation. A process is stationary if its statistical characteristics are the same when examined at any given moment during the time sequence. However, we recently described nonstationarity in rat BP recordings in the form of erratic, large-amplitude events (5). In addition, Roach et al. (21) described temporally localized contributions to human heart rate variability at ultralow frequencies (<0.0033 Hz). It appears that, rather than being stationary, very-low-frequency variability in cardiovascular variables is dominated by temporally localized, large-amplitude events (5, 15, 21, 22). Consequently, conclusions based on standard spectral analysis of cardiovascular variables within the low-frequency range, including that of a weak functional relationship between sympathetic nerve activity and BP, can be misleading (5).
Another very interesting property of cardiovascular signals is that,
when examined in the frequency domain, the power is generally perceived
to increase as a function of 1/f
for very low
frequencies, where
is the slope of the power spectrum curve on a
log-log plot (14, 16). Such a power law relationship between two variables (i.e., power and frequency) is a defining characteristic of a self-similar scaling relationship. Examples of
self-similar scaling that are familiar to everyone include the
branching patterns in tree limbs or the variations in a coast line: the
patterns are similar irrespective of the scale one examines (e.g., a
section of coast a few hundred yards long or a few hundred miles long).
Lack of stationarity in cardiovascular signals could limit the
effectiveness of power spectral processes in the analysis of these
signals. Wavelet analysis, which does not demand that a signal be
stationary (12), is an alternate means to detect the
presence of a scaling relationship. A wavelet is effectively a digital
band-pass filter that can be tuned to any given frequency of interest.
Restated, the wavelet scale can be chosen that specifically selects
those components (i.e., frequencies) in the signal that are of
particular interest. In addition, the wavelet can analyze the frequency
content of the signal over time: one "slides" the wavelet through
the signal across time to determine how power at that frequency
fluctuates from moment to moment. The entire data set can then be
scrutinized to determine how often a fluctuation of a given amplitude
occurred in the signal in the frequency range determined by the scaling
function. This analysis can be performed for any given frequency range
of interest by changing the time scale of the wavelet.
The result of the analysis described above is a probability
distribution for amplitudes of fluctuations over a range of frequencies determined by the time scale chosen to tune the wavelet. One defining characteristic of any self-similar signal is that it conforms to a
"generalized homogeneous function." A function
P(x,
) is a generalized homogeneous function if
it satisfies the following equation
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(1) |
) has the same shape (the
"equal" sign) over a wide range of time scales (i.e., regardless of
the value of L). We found, for example, that the probability
density curves for BP and NT for different frequency ranges (i.e.,
scales) have different widths and different peak locations, but, except
for one unique frequency, all have the same shape or form. In a more
specific sense, Ivanov et al. (12) showed that a
self-similar cardiovascular signal (i.e., heart rate) fits a specific
homogenous function called the gamma distribution. Finally, power law
behavior is a natural consequence of the properties of generalized
homogeneous functions. For example, the gamma distribution is
proportional to the parameter
when plotted as a function of the
dimensionless variable
x (Eq. 4).
In the present experiment, we sought to quantify the relationship between sympathetic NT and BP in the very-low-frequency range. We had several specific objectives: 1) We were intrigued by the isolated, large-amplitude fluctuations that contribute to the very-low-frequency variability. A wavelet analysis was a natural choice to study these temporally localized events, because it does not require that the signal be stationary and it is the best compromise for localizing an event in time and in frequency (6). 2) We computed the cross-wavelet power spectrum (24) between NT and BP to detect the occurrence in time of these large-amplitude, transient events and to determine the frequency range of these events. 3) In addition to these wavelet analyses, we computed a time sequence of cross correlations between NT and BP from overlapping data sets to follow the correlation between NT and BP in time. The combination of a wavelet analysis and a more traditional cross-correlation analysis gave us a consistent picture of the relationship between the sympathetic nervous system and BP in intact animals in the very-low-frequency range and helps explain why standard spectral processes do not detect a strong coherence between these two signals below ~0.1 Hz.
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METHODS |
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Experiments were performed on 23 Sprague-Dawley rats (Harlan Industries, Indianapolis, IN) weighing 330-450 g. The standards for care and use of animals of the American Physiological Society were observed at all stages of the experiment.
Surgery. The rats were anesthetized with pentobarbital sodium (65 mg/kg ip) and prepared for sterile surgery. A Teflon catheter (0.012 ID; catalog no. 30 LW, Small Parts, Miami Lakes, FL) was inserted into the aorta by way of the femoral artery in all 23 animals. In six rats a sympathetic nerve coursing over the aorta toward the kidney was also identified through a flank incision. A small section of this nerve, usually from the cephalad angle formed by the renal artery and aorta, was dissected free of connective tissue and placed on fine, closely spaced (0.4-0.8 mm), bipolar gold electrodes (A-M Systems, Seattle, WA). The exposed nerve and electrode were encased in silicone gel (Wacker Chemie, Munich, Germany). The distal ends of the catheter and wires soldered to the end of the electrodes were tunneled under the skin, exited at the nape of the neck, and led through a flexible tether.
Data acquisition. Data were collected 24 h after surgery in the conscious state while the rat rested quietly in a cage. The arterial pressure signal from a Cobe transducer attached to the femoral artery catheter was amplified and displayed by a Grass model 7 polygraph. The electrical signal from the renal sympathetic nerve was amplified (50,000) and band-pass filtered between 30 Hz and 3 kHz by a Grass P511 differential amplifier.
To obtain accurate measurements from the sympathetic nerve recordings, data were digitized at 10,000 samples/s using a Cache 486 microprocessor and Data Translation DT2821-F analog-to-digital converter. Data were collected for 2-5 h. The initial highly detailed NT signal was full-wave rectified and averaged over every 10 points to produce a 1,000 samples/s signal. This process retains cumulative information from the initial 10,000 samples/s signal. The pressure signals were compressed "on-line" to a 1,000 samples/s signal by saving every 10th point. Because we were interested in the low-frequency range, the data were further compressed to 50 samples/s by averaging every 20 data points. We carefully removed artifacts from the raw time series, using software developed in our laboratory in Visual C++ and saved the edited time series with a sample rate of 500 Hz.Wavelet analysis.
We performed two different types of wavelet analysis. The first,
cumulative variation amplitude analysis, detects the presence of
scaling (12). Cumulative variation amplitude analysis
consists of three steps. In step 1, we calculated the
wavelet transform of the time series. The wavelet transform
allows the study of the signal on any chosen scale
s. The continuous wavelet transform of a time series
x(t) is defined as
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(2) |
has a width on the order of the
scale s and is centered at time t. The Mexican
hat wavelet, which is the second derivative of a Gaussian function, is
orthogonal (i.e., insensitive) to linear trends (12).
Because we were not interested in detecting linear trends in the data,
we used the Mexican hat wavelet in our analysis. For this wavelet,
there is a conversion factor of ~4 between the scale s and
the corresponding Fourier period (24); i.e., applying a
wavelet with a scale s = 0.5 to a time series selects for
rhythms that have a period of ~2.0 s. Using a standard Fourier
transform routine, one can efficiently calculate the convolution
integral in the frequency domain and then take the inverse Fourier
transform to find the wavelet transform in the time domain.
In step 2, we extracted the amplitudes of the variations in
our time series at the frequency range specified by the scale by using
an analytic signal approach (12). Let
c(t) represent an arbitrary signal. The analytic
signal, a complex function of time, is defined by
a(t)
c(t) + s(t), where s(t) is the
conjugate time series. For example, the sine curve is conjugate to a
cosine curve. Then, the amplitude A(t) of the time series is
the magnitude of the complex number a(t); i.e.,
A(t) = 
i. In summary, to
compute A(t), we calculated the wavelet transform
(s,t) and its conjugate time
series by taking the Hilbert transform of
(s,t).
In step 3, we calculated the probability distribution of the
amplitudes A(t) at the specified frequency range for our
time series. After sorting the amplitudes from smallest to largest, we
partitioned the amplitudes into n = 200 bins with equal
numbers of entries to obtain a probability distribution
P(A). To aid in obtaining the peak value
Pmax of P(A), we smoothed the
variations in the experimentally measured curve for P(A) by
applying a second-order Savitsky-Golay filter (18) (with a
window of 17).
The experimentally measured probability distributions were well fit by
the gamma probability density, namely
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(3) |
,
)dx represents the
probability for
events to occur in a time interval x,
where
is the mean rate at which events occur. In our case,
x represents the amplitude of a fluctuation, instead of a
time interval. Possibly, the amplitude of a fluctuation is proportional
to an integration time that is determined by
events. We found that
the probability distributions for fluctuation amplitudes from different
frequency ranges had different
values but the same
. By scaling
the probability distribution by 1/
, one obtains a one-parameter
distribution
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(4) |
x, they all fall onto the same curve. In other words, the probability densities for NT
and BP fluctuations are described by a generalized homogeneous function, which depends on three variables but, after rescaling, depends on only two. Because Pmax is
proportional to
, we can achieve the same effect by scaling by
Pmax. Following Ivanov et al.
(12), we rescaled the probability distribution according to
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(5) |
that minimizes the least-squares residual
error between the fit to the gamma distribution and the scaled
P(A). Because the location of the peak
x0 after scaling is a complicated function of
, it was convenient to simultaneously fit
and
x0.
In a very different application of wavelet analysis, we also calculated
the cross-wavelet power spectrum (24) between NT and BP.
This is an effective way to detect large-amplitude time-localized events. Given two time series x(t) and
y(t), with wavelet transforms
(s,t) and
(s,t), the cross-wavelet
spectrum Wxy(s,t) is
defined as
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*(s,t)
is the complex conjugate of
(s,t). The cross-wavelet
spectrum is complex; therefore, one can define the cross-wavelet power
as |Wxy(s,t)|.
[Because the Mexican hat wavelet is real, we used the Hilbert
transform of
(s,t) as the
complex part of the wavelet transform.]
If it is assumed that both wavelet spectra are
2
distributed with two degrees of freedom (for the real and imaginary
parts of the transform), then significance levels for the cross-wavelet power can be computed from the probability distribution for the square
root of the product of two
2 distributions
(24), namely
|
(6) |
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(7) |
xx(s) is
the average local wavelet power spectrum given by
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(8) |

p' to obtain a threshold Z/2. In other
words, if it is assumed that the time series is a stationary random
process, the probability is less than p that any one of the
values for the scaled cross-wavelet power will exceed Z/2.
Correlations.
In addition to the wavelet analysis, we studied the correlation between
NT and BP in the time domain. To study the correlations between NT and
BP at low frequencies, the data were passed through an eighth-order
Butterworth low-pass filter with a cutoff frequency of 0.2 Hz and
compressed to a sampling rate of 5 Hz. Each time series was then
divided into ~700 segments containing 256 points (or a period of
51.2 s) with 50% overlap. The correlation between NT and BP was
calculated using the fast Fourier transform algorithm on zero-padded
data segments. The correlation in the time domain between lags from
10 to +10 s was computed using the inverse fast Fourier transform. We
displayed the results on a contour plot where the time for each data
segment is defined as the time of the center of the segment.
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RESULTS |
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The physiological NT and BP time series display rhythmic
fluctuations and transient, intermittent fluctuations. Raw time series for NT and BP from rat sds are shown in Fig.
1. The sampling rate for these time
series is 500 Hz. In Fig. 1A, a 5-s interval is displayed.
Pulse synchronous nervous activity is readily apparent during the
occasional dips in BP. Figure 1B shows a 20-s interval from
the same recording. Note the prominent 2.5-s rhythm in BP and the
corresponding bursts in NT. The bursts are roughly 180° out of phase
with the BP fluctuations. Each "burst" of NT in Fig. 1B
corresponds to the increase in NT during the dip in BP in Fig. 1A. Figure 1C shows a 20-min interval from the
same data recording. Note the transient, large-amplitude events in BP
at ~142, 146, 150, 153, and 158 min and the changes in NT
corresponding to these events. The duration of these events is ~40 s.
In the three events at 142, 150, and 158 min, changes in sympathetic
nervous activity lead changes in BP. The other two events have a
different character from these three events (see below).
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Wavelet analysis.
Figure 2A shows the probability (P) of occurrence
of BP fluctuations of a given amplitude (A) in rat sdm. Blue
represents a scale s = 1, which corresponds to BP
fluctuations with a period of ~4 s; red (s = 0.5)
corresponds to ~2 s. For example, the blue distribution shows that
the probability of observing a fluctuation of amplitude 0.8 mmHg within
the ~0.25-Hz range was relatively high, whereas larger and smaller
fluctuations in this frequency range were less likely. In comparison,
the red distribution (~0.4 Hz) was considerably broader, with a peak
that was thereby smaller and less focused. To test for the presence of
scaling, we rescale these probability curves to see if they fall onto a
universal curve (12). We normalize the ordinate against
the peak value for each respective curve
[P(A)/Pmax] and the abscissa by the reciprocal of the same peak value [APmax].
After this rescaling, the curves for the larger scales (i.e., blue,
green, and yellow) are superimposed, whereas the curve for s
= 0.5 (red) is clearly offset. This collapse of the blue, green,
and yellow curves to a common distribution indicates the presence of a
universal scaling in the dynamics underlying the production of the
low-frequency variability in NT and BP. The scale s = 0.5 corresponds approximately to the frequency of the 0.4-Hz rhythm
(3, 4). The presence of the 0.4-Hz rhythm disrupts the
self-similar scaling behavior apparent for the larger scales (i.e.,
s
1.0).
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parameters of the
gamma function to which the data for s = 0.5, 1, 2, 4, 8, 16, and 32 were fit (Eq. 5). The results are shown in Table
1. We ran a one-way, repeated-measures
ANOVA (P < 0.0001 for x0 and
P = 0.003 for
) followed by Bonferroni's post hoc
tests. The post hoc test for x0 (i.e., the
location of the peak for the probability distribution curves; see
Eq. 5) showed a significant difference (P < 0.05) between s = 0.5 and all other scales. However, there
were no other significant differences between scales, indicating
self-similarity between scales ranging from s = 1 to s
= 32. This quantitatively confirms the visual impression in Fig.
2 that the red trace is offset from all
the others. The
post hoc tests (i.e., the general shape of the
probability distribution curve) showed significant differences only
between s = 0.5 and s = 1 and 2.
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(P = 0.773). In
contrast to the construction of Table 1, the values in Table 2 are
calculated by fitting data from all scales, except s = 0.5, rather than a particular scale.
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Time domain analysis.
A contour plot of the correlation in the time domain between NT and BP
for rat sds is shown in Fig.
4. There are strong positive and negative
correlations between NT and BP at certain discrete times. The positive
and negative correlations between 140 and 160 min correspond to the
large-amplitude events detected with the cross-wavelet power spectrum
in Fig. 3. During the strong positive correlations, NT leads BP
(negative lag in Fig. 4). In addition to strong positive correlations,
we find strong negative correlations (e.g., at 146 and 153 min in Fig.
4) where fluctuations in BP lead fluctuations in NT. Such dynamics are
indicative of a biofeedback control system with a functional competency
on the order of tens of seconds (e.g., the baroreflex). The event near 146 min that did not reach statistical significance in the
cross-wavelet analysis shows up here as a relatively weak negative
correlation of about
0.55 between NT and BP. A summary of the results
of our correlation analyses is shown in Table
4. At certain discrete times, we found
strong positive and negative correlations between NT and BP in all six
rats indicative of large-amplitude transient events on time scales
approaching 1 min.
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DISCUSSION |
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Signal analysis of physiological recordings often yields insights as to how the biological system functions or how its function is controlled, which would otherwise go unnoticed. In particular, we (5) and others (15, 21, 22) previously described large-amplitude changes in BP and NT that are transient and occur at irregular intervals. These events were clearly seen to contribute to the very-low-frequency variability in the NT and BP signals, but their physiological significance was not at all clear. Resolving this issue was a particular challenge, because the irregular occurrence (i.e., nonstationarity) of the events precluded the use of "workhorse," spectral analysis tools that are otherwise very powerful. Our analysis circumvented these restrictions and quantified the functional relationship between NT and BP as it changed over time. During many of these transient events, we found a strong correlation between NT and BP that provides, we believe, new insight into how BP is regulated within this very-low-frequency range. Moreover, we found a sharp distinction between the self-similarity that characterized the majority of the lower-frequency signals and fluctuations with periods of ~2 s.
Our time domain analysis (Fig. 4), similar to the cross-wavelet analysis (Fig. 3), revealed that BP and NT were strongly positively correlated during many of the transient events. Because NT leads BP in certain cases, some of these events appear to be produced by nonbiofeedback mechanisms (e.g., central commands), such as we have shown are evoked by an acute behavioral stress (19), or perhaps result from the integration of subtle shifts in blood volume or BP by low-pressure receptors (e.g., atrial receptors). We have reported that these strong positive correlations are absent after transection of the spinal cord between T1 and T2 and, therefore, appear to depend on intact sympathetic outflow to the heart and vasculature (2). In other events, BP leads NT after an inversion, indicative of a feedback control reflex with a time scale of seconds. The baroreflex is the obvious candidate for exerting this latter control. Between these large-amplitude events, the time domain analysis used to construct Fig. 4 suggests that NT and BP are weakly correlated within the frequency range we examined (i.e., at a time scale that far exceeds the usual 2- to 3-beat sequences on which attention typically focuses). It is this apparent poor coupling between these two signals and the mutual "washing out" of the positively and negatively correlated transient events that lead to the lack of coherence between NT and BP that has been reported within the low-frequency range (3).
The wavelet analysis allows a different perspective on the relationship between NT and BP within this low-frequency range. First, the probability distributions for BP with periods that equal or exceed ~4 s (Fig. 2A) "collapse" to virtually a single distribution when rescaled, or normalized, against Pmax (Fig. 2B). [The clear exception to this generalization was the rhythm with a period of ~2 s (see below).] Perhaps even more impressive, however, was the virtual identity between the probability distributions for NT and BP fluctuations within the low-frequency range (Fig. 2C). The describing parameters of gamma distributions to which the BP data were fit were statistically indistinguishable from those to which the NT data were fit (Table 2). This analysis reveals, we believe, a strong functional association between the two signals that is not detected by standard coherence computations (3, 5). The most parsimonious interpretation is that the NT and BP signals are being driven by a common control mechanism. Again, the baroreflex is an obvious candidate, but irrespective of the mechanism, it seems irrefutable that NT and BP are, indeed, functionally coupled at frequencies that are much lower than the 0.1-Hz "cutoff" previously described (5).
These large-amplitude fluctuations are related to the self-similar scaling property that we observed in NT and BP. Surrogate data sets (see METHODS) were constructed by randomly "scrambling" the actual data set (i.e., to produce a "phase-randomized time series") so that the overall content (i.e., the "average of the local wavelet power spectrum") is unchanged but the distinguishing features of the actual data disappear. Most especially, the isolated large-amplitude fluctuations no longer appear in the phase-randomized time series. The resultant probability density curve computed from this surrogate data set is no longer described by a gamma function (data not shown) (12). That is, data sets from which the large-amplitude, transient events are eliminated but have the same average spectrum no longer show the type of self-similar scaling that we observed for NT and BP fluctuations.
The fact that the data conformed to a generalized homogeneous function, i.e., the gamma function, is associated with power law behavior, which, in turn, is characteristic of scaling (see the introduction). Physically, scaling relations arise when the underlying dynamics "look" the same, irrespective of the time scale of the analysis (10). In sharp contrast, the characteristics of signals with strong rhythmicity absolutely depend on the time scale chosen for their analysis. It follows, therefore, that because we observed self-similar scaling, the dynamics underlying the production of these large-amplitude events occur over a range of time scales; i.e., there is no one characteristic time scale, as is the case for rhythmic fluctuations. Although isolated large-amplitude events dominate very-low-frequency variability (0.001-0.1 Hz), feedback oscillations dominate the low-frequency range (0.1-1.0 Hz) (4, 20). The probability distribution centered around a period of 2 s (i.e., 0.4 Hz; red trace in Fig. 2, A and B) was clearly offset from all the curves describing the lower frequencies. We believe that this change from self-similar to rhythmic fluctuations explains the fundamentally different statistical characteristics of the fluctuations with a 2-s period.
In reference to Table 2, our values for
from NT and BP fluctuations
compare very well with the findings of Ivanov et al. (12),
who reported
= 2.8 and 2.4 for daytime and nighttime heart
rate variability, respectively, in humans. (Their parameter
is
equal to our
1.) We found a significant deviation between the tail of the gamma distribution and the universal probability distribution from the data; i.e., large-amplitude events are more probable in the data than would be expected from a true gamma distribution. Again, this discrepancy for the probability distributions of NT and BP fluctuations is in agreement with the observations of
Ivanov et al. for the nighttime heart rate variability.
Perspectives
Perhaps, sympathetic regulation of the cardiovascular system may be in part through large-amplitude, transient events (Fig. 1C). The presence of large-amplitude, transient events in NT indicates the cooperative behavior of a large number of neurons. Moreover, the presence of a universal scaling law in our data indicates that the cooperative dynamics may be governed by a single physical process with no special time scale. The emergent behavior of large populations of neurons may occur on a range of scales that is described by the renormalized dynamics (10) of interaction between single neurons. Possibly, the large-amplitude events resulting from the dynamics of large populations of neurons are due to the integration of information from the cardiovascular system over a range of time scales. These events may be coordinated in parasympathetic and sympathetic activity to the heart and to the vasculature to control BP.| |
ACKNOWLEDGEMENTS |
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This work was supported by American Heart Association Beginning Grant-in-Aid (Ohio Valley Affiliate) 9806307 to D. E. Burgess and National Institute of Neurological Disorders and Stroke Grant NS-39774 to the University of Kentucky.
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FOOTNOTES |
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Address for reprint requests and other correspondence: D. E. Burgess, Dept. of Chemistry and Physics, Asbury College, Wilmore, KY 40390-1198 (E-mail: deburgess{at}asbury.edu).
The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
First published October 3, 2002;10.1152/ajpregu.00002.2002
Received 7 January 2002; accepted in final form 21 September 2002.
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