Assessing erectile neurogenic dysfunction from heart rate variability through a Generalized Linear Mixed Model framework.

BACKGROUND: The low (LF) vs. high (HF) frequency energy ratio, computed from the spectral decomposition of heart beat intervals, has become a major tool in cardiac autonomic system control and sympatho-vagal balance studies. The (statistical) distributions of response variables designed from ratios of two quantities, such as the LF/HF ratio, are likely to non-normal, hence preventing e.g., from a relevant use of the t-test. Even using a non-parametric formulation, the solution may be not appropriate as the test statistics do not account for correlation and heteroskedasticity, such as those that can be observed when several measures are taken from the same patient. OBJECTIVES: The analyses for such type of data require the application of statistical models which do not assume a priori independence. In this spirit, the present contribution proposes the use of the Generalized Linear Mixed Models (GLMMs) framework to assess differences between groups of measures performed over classes of patients. METHODS: Statistical linear mixed models allow the inclusion of at least one random effect, besides the error term, which induces correlation between observations from the same subject. Moreover, by using GLMM, practitioners could assume any probability distribution, within the exponential family, for the data, and naturally model heteroskedasticity. Here, the sympatho-vagal balance expressed as LF/HF ratio of patients suffering neurogenic erectile dysfunction under three different body positions was analyzed in a case-control protocol by means of a GLMM under gamma and Gaussian distributed responses assumptions. RESULTS: The gamma GLMM model was compared with the normal linear mixed model (LMM) approach conducted using raw and log transformed data. Both raw GLMM gamma and log transformed LMM allow better inference for factor effects, including correlations between observations from the same patient under different body position compared to the raw LMM. The gamma GLMM provides a more natural distribution assumption of a response expressed as a ratio. CONCLUSIONS: A gamma distribution assumption intrinsically models quadratic relationships between the expected value and the variance of the data avoiding prior data transformation. SAS and R source code are available on request.

Assessment of cardiovascular autonomic control by the empirical mode decomposition.

OBJECTIVES: Heart-rate variability (HRV) is an interesting tool for assessing cardiac autonomic system control, but nonstationarities raise problematic issues. The objective of this paper is to show that adapted signal processing tools may cope with nonstationary situations and improve the analysis of HRV. METHODS: We propose to use the recent method of Empirical Mode Decomposition (EMD), so as to analyze the cardiac sympatho-vagal balance on automatically extracted modes. The method, which is fully data-adaptive, consists in an iterative decomposition based on the idea that any signal can be locally represented as an oscillation superimposed to a more regular trend. When a signal is composed of distinct nonstationary components, EMD therefore achieves a time-varying filtering which effectively separates them. RESULTS: The method has been applied to situations where postural changes occur, provoking instantaneous changes in heart rate as a result of autonomic modifications. In the considered application where the sympatho-vagal balance is quantified by comparing the low-frequency (LF) and high-frequency (HF) components of RR intervals, EMD automatically achieves a separation of these components upon which further processing can be carried. Visualizing the decomposition in the time-frequency plane, we can identify local events due to the postural changes, and we can assess a (time-varying) HF vs. LF discrimination without resorting to some fixed high-pass/low-pass filtering. CONCLUSION: Assessing cardiovascular autonomic control by resorting to LF/HF measurements may prove difficult in nonstationary situations where the use of a priori fixed filters can be questioned. Because it is both local and fully data-adaptive, EMD appears as an appealing and versatile pre-processing technique for overcoming some of the limitations that conventional spectral methods are faced with in nonstationary situations.

Smoothed pseudo Wigner-Ville distribution as an alternative to Fourier transform in rats.

Techniques for examining signals in the time and frequency domains are well-established tools. These tools have their limitations; they tell us in a broad sense where the signal component exists in the frequency domain, but they do not tell us how its frequency characteristics change over time. The time-frequency has become a powerful alternative for the analysis of signals. Among various time-frequency distribution methods, one of the most studied is the Wigner-Ville distribution. The aim of this study was to evaluate in conscious rats smoothed pseudo Wigner--Ville distribution (SPWVD) as an alternative to the fast Fourier transform (FFT) in RR intervals and in systolic blood pressure (SBP), before and after adrenergic and cholinergic receptor blockade. Fourteen Wistar rats equipped with telemetry probe were evaluated: (1) under control conditions; (2) after injection of saline (100 microl kg(-1) i.v.); (3) after atenolol (1 mg kg(-1) i.v.); (4) after atropine methyl nitrate (0.5 mg kg(-1) i.v.); and (5) after phentolamine (5 mg kg(-1) i.v.). FFT and SPWVD were applied to RR intervals and SBP time series. Six-minute time series of RR intervals, systolic and diastolic pressures were analysed. The bias and distribution of differences between FFT and SPWVD methods in RR intervals under base conditions were 1.4+/-0.4% (r2=0.94; P<0.01) in LF/LF+HF: 1.5+/-0.5% ( r2=0.92; P<0.01) in HF/LF+HF and 4.8+/-1.9% (r2=0.92; P<0.01) in LF/HF. In SBP the bias and distribution were 1.5+/-0.8% (r2=0.90) P<0.05) in LF/LF+HF and 1.7+/-0.6% (r2=-0.92; P<0.01) in HF/LF+HF. In the frequency domain analysis of RR intervals and SBP there was no difference between FFT and SPWVD. The agreement between the methods demonstrates that in stationary signals both methods can be used interchangeably. SPWVD may be an interesting tool to analyse biomedical signals; it provides a good resolution at high frequency and a good frequency resolution at low frequencies independently if signals remain stationary.

No effect of venoconstrictive thigh cuffs on orthostatic hypotension induced by head-down bed rest.

Orthostatic intolerance (OI) is the most serious symptom of cardiovascular deconditioning induced by head-down bed rest or weightlessness. Wearing venoconstrictive thigh cuffs is an empirical countermeasure used by Russian cosmonauts to limit the shift of fluid from the lower part of the body to the cardio-cephalic region. Our aim was to determine whether or not thigh cuffs help to prevent orthostatic hypotension induced by head-down bed rest. We studied the effect of thigh cuffs on eight healthy men. The cuffs were worn during the day for 7 days of head-down bed rest. We measured: orthostatic tolerance (stand tests and lower body negative pressure tests), plasma volume (Evans blue dilution), autonomic influences (plasma noradrenaline) and baroreflex sensitivity (spontaneous baroreflex slope). Thigh cuffs limited the loss of plasma volume (thigh cuffs: -201 +/- 37 mL vs. control: -345 +/- 42 mL, P < 0.05), the degree of tachycardia and reduction in the spontaneous baroreflex sensitivity induced by head-down bed rest. However, the impact of thigh cuffs was not sufficient to prevent OI (thigh cuffs: 7.0 min of standing time vs. control: 7.1 min). Decrease in absolute plasma volume and in baroreflex sensitivity are known to be important factors in the aetiology of OI induced by head-down bed rest. However, dealing with these factors, using thigh cuffs for example, is not sufficient to prevent OI. Other factors such as venous compliance, microcirculatory changes, peripheral arterial vasoconstriction and vestibular afferents must also be considered.

Effect of acute atenolol on short-term blood pressure variability and baroreflex sensitivity in rats.

Beta-blocker therapy for hypertension or coronary artery disease is common, but there are a lot of controversies about its effects on short-term blood pressure variability and arterial baroreceptor reflexes. The aim of this study was to evaluate the effects of acute atenolol on baroreflex sensitivity (BRS) and on the spontaneous variability of systolic blood pressure (SBP) and RR intervals in conscious rats. Ten Wistar rats equipped with telemetry system were evaluated: 1) under control conditions; 2) after injection of saline; and 3) during beta1-adrenergic blockade by atenolol. Fast Fourier transform analysis was applied to RR intervals and SBP. Atenolol increased RR intervals significantly by 14% and the variation coefficient of the RR intervals by 31%. SBP was reduced significantly by 9%. In frequency domain, beta1-blockade in RR intervals increased very low frequency by 33% and the total power by 22% and decreased low frequency by 25%. The ratio of low to high frequency power decreased by 60%. Frequency domain variables in SBP were not significantly changed after beta1-adrenergic receptor blockade. BRS (gain alpha) was not significantly altered by beta-blockers. Acute atenolol decreased SBP and increased RR intervals with no change in BRS, indicating 'resetting' of baroreflex function.

Even slight movements disturb analysis of cardiovascular dynamics.

We hypothesized that spontaneous movements (postural adjustments and ideomotion) disturb analysis of heart rate and blood pressure variability and could explain the discrepancy between studies. We measured R-R intervals and systolic blood pressure in nine healthy sitting subjects during three protocols: 1) no movement allowed, 2) movements allowed but not standing, 3) movements and standing allowed. Heart rate and blood pressure were not altered by movements. Movements with or without standing produced a twofold or greater increase of the overall variability of R-R intervals and of the low-frequency components of spectral analysis of heart rate variability. The spectral exponent beta of heart rate variability (1. 123 at rest) was changed by movements (1.364), and the percentage of fractal noise (79% at rest) was increased by standing (91%, coarse-graining spectral analysis). Spontaneous movements could induce a plateau in the correlation dimensions of heart rate variability, but they changed its nonlinear predictability. We suggest that future studies on short-term cardiovascular variability should control spontaneous movements.

Regional blood flow in conscious rats after head-down suspension.

Exposure to microgravity in humans causes cardiovascular deconditioning affecting blood pressure, heart rate and vascular responsiveness. This study investigated cardiac output, arterial blood pressure and regional blood flows [radioactive microspheres: 57Co, 15.5 (SEM 0.1) microm in diameter] in conscious and freely moving rats subjected to 14 days of simulated microgravity (head-down suspension, HDS) in male Wistar rats: control (horizontally attached, n = 7), suspended for 14 days (n = 8) and suspended/allowed to recover for 10 min (R10min, n = 5) or 24 h (n = 9). Compared to the control group, 14 days of HDS resulted in reduced total peripheral resistance (37%); an increased cardiac index (65%) was associated with no significant change in the mean arterial pressure BPa. There were elevated brain (63%), visceral (> 20%), hindlimb (> 80%) and forelimb (> 215%) muscle blood flows. In the R10min group, the BPa decreased (18%) and the regional blood flows returned to control values. Within 24 h the BPa as well as cardiac index and total peripheral resistance were restored. In conclusion, 14 days of HDS engendered local circulatory changes resulting in transient blood pressure instability during recovery.

Recording heart rate and blood pressure in rats during parabolic flight.

The way in which the cardiovascular system adapts to weightlessness is still under discussion. No data are yet available on the responses of rats during space flight, although this animal is commonly used in simulation studies. We have designed and tested a protocol to study the short term responses of the cardiovascular system to weightlessness during parabolic flight. A telemetry system was used to measure heart rate (HR) and blood pressure. It was possible to collect and record radio-signals without any interference. Microgravity caused a reduction in HR, an increase in mean arterial pressure (MAP, 7%), and a non-significant decrease in central venous pressure (CVP, 13%). The change in CVP was similar to the decrease observed in human space flight. This type of study may also be feasible for longer exposure of rats to microgravity (space flight).

A metabolism study of human masseter muscle by 31P magnetic resonance spectroscopy during long periods of exercise and recovery.

The metabolism of the human masseter muscle was investigated using phosphorus (31p) magnetic resonance spectroscopy (MRS) during long periods of exercise and recovery. Eleven subjects aged 19 to 28 yr were examined by 31p MRS during four consecutive periods of 13 min each: rest, exercise, recovery 1 and 2. For each subject, a biting force equal to 20% of maximum voluntary biting force was applied and controlled during the exercise period to produce maximum fatigue. 31p MR spectra were localized from a 24 cm3 volume of interest using an image selected in vivo spectroscopy (ISIS) sequence and a 6 cm diameter surface coil placed on the left masseter. Compared to the resting level, the phosphocreatine (PCr) content decreased by 26% during exercise, while the inorganic phosphate (Pi) concentration increased by 65%. During the two recovery periods, the Pi content remained decreased compared with the resting level by 36% and 30%, respectively. The Pi/PCr ratio was increased from 0.30+/-0.04 at rest to 0.63+/-0.13 during exercise while the adenosine triphosphate (ATP)/Pi ratio was decreased. The pH decreased from 7.02+/-0.03 to 6.93+/-0.04 during exercise and returned to control level (7.09+/-0.08) only during the second recovery period. These results suggest that the masseter muscle is characterized by high ATP turnover and, therefore, high oxidative phosphorylative activity in agreement with its constitution of predominantly fatigue resistant type I fibers.

Cardiovascular adaptations to parabolic flight in rats: a radio-telemetry feasibility study.

This experiment was a feasibility study which consisted in investigating arterial blood pressure and heart rate to transient and repeated exposure to microgravity in eight unrestrained rats previously implanted with radio-telemetry transmitter. The aim was to perform such recordings throughout all the phases of a parabola during parabolic flights. This study revealed that it was possible to collect the radio-signal without any interference with electronic or magnetic environment. We observed in microgravity a significant reduction in heart rate (6%) and a significant increase in arterial blood pressure (7%). In conclusion, such a study seems to be feasible during longer exposure to microgravity (space flight) in order to study the cardiovascular adaptation in rat.

[The demonstration of a relation between premolar force and molar force within the limits of a simplified approach to masticatory system functioning]. / Mise en évidence d'une relation entre la force prémolaire et la force molaire dans le cadre d'une approche simplifiée du functionnement de l'appareil manducateur.

The purpose of this study is to show the existence of a correlation between the premolar and molar clenching forces obtained during a voluntary clenching exercise. The study concerned 32 volunteers aged 21 to 28 with no manducatory problems. The forces were obtained using a device with four Kiowa traducers arranged in a complete Wheatstone bridge. The analysis of the results obtained showed that there was a positive correlation between maximum molar and premolar. The analysis also shows that three results are a direct application of the theory of momentum. This enables us to propose a simple biophysical model to explain the functioning of the masticatory system.

[The determination of the intensity of premolar and molar maximal forces during the isometric contraction of the masticatory muscles due to forced mandibular closure]. / Détermination de l'intennsité des forces maximales prémolaires et molaires au cours de la contraction isométrique des muscles masticateurs par fermeture forcée de la mandibule.

The purpose of this study to quantify the activity of the masticatory muscles. For this we recorded the maximum bite force of opposing teeth during voluntary clenching. The study was carried out on 31 students aged 21 to 28 with a satisfactory dental arch. The measurements were obtained on one side only. Nine thickness of the traducers were used for the premolar zone and two for the molar zone. We determined an optimal thickness enabling the highest measurement: 6.5 mm for the premolars and 5.2 for the molars. The highest measurements were averaged and the following results were obtained: premolars-35 kgf for females and 50 kgf for males; molars 100 kgf for females and 130 kgf for males.

Soleus muscle alterations in spontaneously hypertensive rats are not dependent on activation of beta 2-adrenergic receptors.

The effects of selective beta 2-adrenergic blockade with ICI 118,551 on the histochemical, biochemical, and contractile properties of slow-twitch soleus muscle from spontaneously hypertensive (SHR) and normotensive (WKY) rats were examined from birth to 8-10 weeks of age. Chronic treatment of normotensive rats with ICI 118,551 caused an impairment in the differentiation of slow type fibers during development but failed to alter the fiber type distribution of hypertensive rats. beta 2-Adrenoreceptor blockade was ineffective in reducing the enhanced glycolytic and oxidative capacities of soleus in hypertensive rats. The suggestion can be made that beta 2-adrenoreceptor activation is not responsible, at least directly, for the histochemical and biochemical alterations of slow muscle from hypertensive rats.

Fasting-induced rise in locomotor activity in rats coincides with increased protein utilization.

The aim of this study was to investigate the possible relation between the modifications in locomotor activity (on running wheel) which occur during prolonged fasting and changes in the utilization of energy reserves. In 18-week-old rats, we found that the rate of body mass loss reflects the changes in nitrogen excretion that occur over three phases of fasting: (I) initially decreasing, (II) maintained at a low level and (III) increasing. Locomotor activity started to increase during phase II without a change in its nycthemeral pattern. By contrast, the 10-fold higher daily locomotor activity that occurred in phase III was marked by a higher proportion of diurnal activity. Using 9-, 18-, and 33-week-old rats, in order to obtain a different timing in the metabolic changes during fasting, we could confirm the coincidence between the later rise in locomotor activity and the occurrence of phase III. Refeeding of rats of either age in phase III rapidly suppressed fasting-induced changes in locomotor activity. These data accord with the idea that behavioral changes reflecting the search for food are triggered by a later and reversible change in the utilization of body protein vs. lipid stores during prolonged fasting.

Structural and metabolic properties of rat muscle exposed to weightlessness aboard Cosmos 1887.

Male Wistar rats were subjected to 12.5 days of weightlessness aboard Cosmos 1887. Histomorphometric and biochemical analyses were investigated in soleus (SOL), plantaris (PL) and extensor digitorum longus (EDL) muscles of flight rats (group F) and compared with data from two groups of terrestrial controls: one group living free in a vivarium (group V) and another subjected to a flight simulation except for the state of weightlessness (group S). Relative to groups V and S, no alteration in the percentage distribution of fibres had occurred in SOL, PL or EDL, after the flight. In SOL muscles from group F animals, cross-sectional areas of all fibre types were reduced to a greater extent (-40%) than capillary to fibre ratio (-24%) leading to a higher capillary density (+33%) than in V and S groups. In PL, type I, IIA and IIB fibre cross-sectional areas were less decreased (-25%). In EDL, only fast-twitch fibre cross-sectional areas showed an average decrease of 30%. Capillary per fibre ratio was reduced by 15% and 28% respectively in PT and EDL muscles from group F rats compared to control groups V and S. Citrate synthase and 3-hydroxyacyl-coenzyme A dehydrogenase activities remained unchanged in SOL, PL and EDL following spaceflight. These findings indicate greater atrophy and functional alterations (capillarity) compared to those observed after 7 days of microgravity on Cosmos 1667.

Effect of almitrine administration on pulmonary arterial pressure in resting and exercising dogs.

In addition to its well-known ventilatory effect, a small rise in pulmonary arterial pressure or pulmonary vascular resistance is occasionally observed with chronic administration of almitrine. In order to test the hypothesis of enhancement of exercise pulmonary vasoconstriction by almitrine, mongrel dogs were studied at rest and during submaximal exercise before and after 4 weeks of chronic ingestion of almitrine (10 mg/kg). It was shown that resting pulmonary arterial pressure (PAP) remained unchanged by almitrine treatment. However, when exercise was superimposed on almitrine medication, PAP was significantly increased throughout the exercise bout. Thus, the rise in PAP during the 20th min of exercise averaged 8.7 +/- 3.4 mm Hg after almitrine treatment while PAP increased by only 1.3 +/- 1.7 mm Hg before medication. The exaggerated exercise-induced PAP response in conjunction with the enhanced secretion of norepinephrine that we observed during almitrine treatment suggests that catecholamine could be involved in the pulmonary haemodynamic adjustments. Furthermore, mixed-venous PO2 (PvO2) both during rest and exercise declined with the prolongation of almitrine ingestion, suggesting that PvO2 might possibly be implicated in the pulmonary haemodynamic response to almitrine, in the same way as it is involved in the hypoxia-induced pulmonary vasoconstriction. These findings demonstrate that almitrine medication, even at a high dose, does not have any deleterious effect on pulmonary vasculature in resting conditions, but prolonged submaximal exercise should be proscribed in patients on a long-term therapy.

Effects of swim training alone and in combination with clonidine and rilmenidine on blood pressure, plasma electrolytes, vasopressin, and renin activity in spontaneously hypertensive rats.

Adaptations to the effects of clonidine (CL) and rilmenidine (R) were studied during a 12-week training program (swimming) in spontaneously hypertensive rats (SHR) and Wistar-Kyoto (WKY) rats. Systolic blood pressure (SBP) was regularly measured during this period. Body weight (BW) was determined at the beginning and at the 12th week. Plasma parameters, cardiac determinations, vasopressin (pAVP), and plasma renin activity (PRA) were measured only at the end of the experiment. Both SBP and ponderal benefit were reduced by CL, R, and training. Contrary to beta-adrenoceptor blocking agents, we found no inhibition of the beneficial effect on SBP of training in combination with CL or R. Plasma and hypothalamic vasopressin were reduced by both drugs but only CL increased plasma renin activity (PRA) although its mechanisms of action are still not clearly understood. Our results suggest that CL and R as well as swim training can be considered as an effective countermeasure in SHR. Moreover, the heterogeneity of action of CL and R on some of the parameters tested is in favor of different pharmacological properties for these drugs.

Skeletal muscle adaptation to physical training and beta-adrenergic blockade in spontaneously hypertensive rats.

The effects of training alone or in combination with long-term, non-selective, beta-adrenergic blockade on histochemical and biochemical properties of fast-twitch [extensor digitorum longus muscle (EDL)] and slow-twitch [soleus muscle (Sol)] muscle were analyzed in spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto strain rats (WKY). Fiber type distribution of Sol was drastically modified in SHR with fewer type I fibers and more type IIA fibers. No such histochemical alterations were observed in EDL. While prolonged swimming training remained ineffective in inducing both histochemical and biochemical improvement in WKY, SHR displayed a significant enhancement of capillarization and oxidative capacity in both Sol and EDL. However, in long-term beta-blocks rats training failed to improve significantly the oxidative capacity of SHR muscles, suggesting that beta-adrenoreceptor stimulation is necessary for a fully efficient adaptation of muscular metabolism to physical training.

Effect of spontaneous recovery or retraining after hindlimb suspension on aerobic capacity.

The purpose of this study was to compare the effects of spontaneous recovery or recovery by treadmill training (180 min/day, 5 days/wk, 30 m/min for 8 wk) on maximal O2 uptake (VO2max), histochemical and biochemical muscular properties (soleus), of rats subsequent to 5 wk of hindlimb suspension. Spontaneous recovery reversed the 15% reduction in VO2max, whereas training posthypokinesia induced a 20% increase over control values. In the spontaneous recovery group, both citrate synthase and 3-hydroxyacyl-CoA dehydrogenase activities, decreased by hypokinesia (-40%), increased but remained 20% below the control level. In the training posthypokinesia group, an increase of these activities over control occurred (+50 and +20%, respectively). Recovery or training led to a 100% type I distribution in soleus muscle and to a recovery of all fibers' cross-sectional areas. In the spontaneous recovery group, capillaries per fiber, decreased by 46%, returned to the normal range. In the training posthypokinesia group, training induced an increase in capillaries per fiber above their control values (+23%). These results point to the plasticity of the muscle and indicate the necessity of a posthypokinesia training program for recovery of the total oxidative enzyme capacity.