Interpretation of drug concentrations in an alternative matrix: the case of meprobamate in bile.

Investigating toxicological causes of death may require alternative matrices when the usual ones are lacking. Whereas forensic toxicology uses bile almost only for xenobiotic screening, a diagnostic test interpreting postmortem bile concentrations of meprobamate is reported. Based on 128 sets of autopsy data, its intrinsic qualities were good, with 0.95 sensitivity and 0.93 specificity. In a French forensic population, the positive and negative predictive factors were 0.90 and 0.97, respectively. It is a useful means of revealing overdoses where blood samples are not available or of confirming blood tests when postmortem redistribution is suspected.

Noninvasive exploration of cardiac autonomic neuropathy. Four reliable methods for diabetes?

OBJECTIVE: The purpose of this work was to assess relevant information that could be provided by various mathematical analyses of spontaneous blood pressure (BP) and heart rate (HR) variabilities in diabetic cardiovascular neuropathy. RESEARCH DESIGN AND METHODS: There were 10 healthy volunteers and 11 diabetic subjects included in the study. Diabetic patients were selected for nonsymptomatic orthostatic hypotension in an assessment of their cardiovascular autonomic impairment. Cardiac autonomic function was scored according to Ewing's methodology adapted to the use of a Finapres device. The spontaneous beat-to-beat BP and HR variabilities were then analyzed on a 1-h recording in supine subjects. The global variabilities were assessed by standard deviation, fractal dimension, and spectral power. The cardiac baroreflex function was estimated by cross-spectral sequences and Z analyses. RESULTS: In diabetic patients, Ewing's scores ranged from 1 to 4.5, confirming cardiovascular autonomic dysfunction. In these diabetic patients, global indices of variabilities were consistently lower than in healthy subjects. Furthermore, some of them (standard deviation and fractal dimension of HR, spectral power of systolic blood pressure and HR) were significantly correlated with the Ewing's scores. The Z methods and the spectral analysis found that the cardiac baroreflex was less effective in diabetic subjects. However, the baroreflex sensitivity could not be reliably assessed in all the patients. The sequence method pointed out a decreased number of baroreflex sequences in diabetic subjects that was correlated to the Ewing's score. CONCLUSIONS: Indices of HR spontaneous beat-to-beat variability are consistently related to the degree of cardiac autonomic dysfunction, according to Ewing's methodology. The Z method and spectral analysis confirmed that the cardiac baroreflex was impaired in diabetic patients. These methods might be clinically relevant for use in detecting incipient neuropathy in diabetic patients.

Mental stress-induced increase in blood pressure is not related to baroreflex sensitivity in middle-aged healthy men.

The baroreflex that acts to blunt blood pressure (BP) variations through opposite variations in heart rate should limit the BP increase produced by an emotional challenge. However, relations between baroreflex sensitivity and BP reactivity induced by a psychological stress in a large group of adults have never been firmly established. In 280 healthy men, rest (10 minutes) and stress (5 minutes) BP and heart rate were recorded beat to beat by a blood pressure monitor. The mental stress was elicited by a well-standardized computerized version of a word color conflict stress test (Stroop Color Test). Rest and stress baroreflex sensitivity was assessed by the cross-spectral analysis of BP and heart rate and by the sequence method. The stress-induced increase in systolic BP (22.4+/-0.1 mm Hg) was not correlated with resting baroreflex sensitivity but was slightly correlated (r=0.2, P<0.001) with BP variability assessed either by standard deviation or by mid-frequency band spectral power. Our results suggested that a centrally mediated sympathetic stimulation overcame cardiac autonomic regulation and emphasized the role of the sympathetic vasoconstriction in the pressure response at the onset of the stressing stimulation. During the sustained sympathoexcitatory phase, the cardiac baroreflex blunts BP variations but at a lower sensitivity.

Spontaneous cardiac baroreceptor reflex and regional circulations in conscious rats.

OBJECTIVE: To determine the relationships between the activity of the spontaneous cardiac baroreceptor reflex (BRX) and regional vascular dynamics in conscious unrestrained rats. DESIGN: Simultaneous beat-to-beat recordings of blood pressure and measurements of indices of mesenteric and hindquarters blood flows were performed in seven male normotensive conscious rats under baseline conditions during 1 h. METHODS: The relationships between systolic blood pressure (SBP) and heart rate were examined using a computerized method, giving the degree of statistical dependence between values of the two parameters with the Z coefficient. This analysis permitted detection of cardiac beats related to cardiac BRX activity. Z analysis was also applied to SBP and to mesenteric or hindquarters blood flow or vascular resistance. Time-series representations were used to describe the chronological patterns corresponding to BRX spontaneous activity. RESULTS: In these rats, which presented spontaneous BRX-like patterns manifested by the Z analysis, BRX was active during approximately 15% of the time. For 80% of these patterns, cardiac BRX acted to buffer falls in SBP. These depressor events were preceded by muscular vasodilations whereas vasodilations in the mesentery were delayed. Z analysis of these spontaneous haemodynamic patterns confirmed that muscular vasodilations were strongly associated with falls in SBP, whereas vasodilations lagged behind SBP by about seven beats and were not dependent on simultaneous SBP values. CONCLUSIONS: The Z method showed that spontaneous hindquarters vasodilations were associated with cardiac BRX patterns and that mesenteric vasodilations were not concomitant with falls in SBP and may be the consequence either of a delayed central response or of a local mechanism such as myogenic autoregulation.

Heterogeneous computer network for real-time hemodynamic signal processing.

A computer network is described that allows real-time processing, graphical monitoring and off-line analysis of blood pressure, nervous activity and Doppler signals recorded in conscious rats. Real-time processing is performed by an acquisition station using a powerful microprocessor, allowing extraction and storage of several characteristic parameters from each cardiac cycle and real-time graphical monitoring. The experimenter can thereby follow the time evolution of the hemodynamic parameters. Experimental data are sent through the local network to a workstation that ensures off-line processing such as chronograms, histograms and statistical analyses.

[Role of sympathetic nervous system on blood pressure and heart rate variabilities in the rat: spectral analysis]. / Rôle du système nerveux sympathique dans la variabilité de la pression artérielle et de la fréquence cardiaque chez le rat: étude par analyse spectrale.

The role of the sympathetic nervous system (SNS) in the genesis of the beat-to-beat variability of blood pressure (BP) and heart rate (HR) was studied using spectral analysis. The SNS activity of genetically normotensive rats of the Lyon strain (LN) was abolished in 2 ways: 1) adrenergic blockade with administrations of phentolamine (PHE, 5 mg/kg i.v.) or propranolol (PRO, 5 mg/kg i.v.) or both drugs (PHEPRO) or 2) destruction of the sympathetic fibers with an early chronic treatment with guanethidine (LNSx). HR and systolic (SBP) and diastolic (DBP) BP spectra were computed using 50 to 100 stationary sequences from 1 h beat-to-beat recordings and covered the 0.015-3.85 Hz frequency band. In LN rats, spectra obtained in baseline conditions exhibited a high-frequency peak (HF, 1.04-1.13 Hz) related to the respiratory frequency and a mid-frequency peak (MF, 0.38-0.45 Hz). Concerning BP, the alterations of the SNS induced significant decreases of the total modulus (mmHg/Hz1/2) of the MF band (0.27-0.74 Hz): [formula: see text] In addition, the MF peak of SBP and DBP spectra was abolished in LNSx and PHEPRO rats. Concerning HR spectra, the moduli of all the frequency components were decreased in PHEPRO rats only. In conclusion, the similarity of the results observed in LNSx and PHEPRO rats shows that the MF oscillations of BP depend in a large extent (65%) on the action of the sympathetic fibers and involve the activation of both alpha- and beta-adrenoceptors. Concerning DBP, this effect is especially mediated by the activation of alpha-adrenoceptors.(ABSTRACT TRUNCATED AT 250 WORDS)

[Computerized analysis of cardiovascular activity in the conscious unrestrained rat. Application to the study of a new antihypertensive agent]. / Analyse informatisée de l'activité cardiovasculaire chez le rat conscient et non restreint. Application à l'étude d'un nouvel antihypertenseur.

A more precise knowledge of cardiovascular activity in rats needs its direct recording for long periods of time, in conscious unrestrained animals. A system has been developed, which allows direct recording of blood pressure (BP) in freely moving rats, by means of a floating catheter chronically inserted in the abdominal aorta, connected to a rotating swivel, and then to a pressure transducer. A mini-computer performs on-line processing of BP curves, and stores the values of 5 cardiovascular parameters on hard disk: systolic, diastolic and mean arterial pressures, heart rate and dp/dt max. Off line processing allows graphic and statistical analysis of the data. Simultaneously recording from two rats is very useful in pharmacology because a treated animal and a control one are studied at the same time and in the same conditions. This methodology was used to evaluate the effects of a chronic treatment (2 mg/kg, ip, during 8 days) and of single injections (0.5 and 3 mg/kg, iv) of a new antihypertensive substance, CM 40441a, in 15 W old males SHR.

[The sympathetic and renin-angiotensin systems are complementary in blood pressure regulation]. / Les systèmes sympathique et rénine angiotensine se complètent pour contrôler la pression artérielle.

The consequences of a chronic destruction of sympathetic nerves or of the blockade of the renin angiotensin system (RAS) on the blood pressure (BP) level and its spontaneous variability were studied using a computerized method which allows a continuous monitoring of BP in conscious unrestrained rats. Guanethidine, used to lesion the sympathetic fibres did not alter the BP level but significantly enhanced its variability. On the opposite, angiotensin converting enzyme inhibition decreased the BP level without changing its variability. It is concluded that both systems play a complementary role as the RAS determines the long-term BP level while the sympathetic nerves control its short-term variability in conscious animals.

Antihypertensive effects of S9490-3, a new converting enzyme inhibitor, in the conscious spontaneously hypertensive rat.

The acute and chronic cardiovascular effects of S9490-3, a new converting enzyme inhibitor, were studied by means of a computerized analysis of the intra-aortic blood pressure recorded for 48 consecutive hours, in the conscious, unrestrained adult spontaneously hypertensive rat (SHR). The software used allowed the simultaneous study of one control and one treated animal. A single intravenous dose of S9490-3 (2 mg/kg) induced a rapid fall in systolic and diastolic blood pressure, which remained significant for 11 h. This decrease was not associated with an elevation in heart rate. A chronic oral treatment with S9490-3 (1 mg/kg/24 h for 12 days) significantly lowered systolic and diastolic blood pressure and reduced the variability of systolic blood pressure. The heart rate was slightly increased at rest (day time) but significantly elevated in the freely exercising SHR (night time). However, it must be noted that this increase remained moderate. The baroreflex sensitivity of chronically treated animals was enhanced when measured after phenylephrine but not after nitroglycerine bolus. Chronically given S9490-3 significantly reduced the pressor effect of phenylephrine and decreased the left ventricle weight.

Computer analysis of intraarterially recorded blood pressure in conscious unrestrained rats.

A precise knowledge of cardiovascular activity and its pharmacological modifications in the conscious unrestrained animal requires intraarterial blood pressure recording for long periods of time. For that purpose, a technique was developed involving chronic catheterization of the rat abdominal aorta and the use of a computer system to perform the digitization of the blood pressure signal and the on-line calculation of five cardiovascular parameters. By means of duplexing, two rats (one treated and one control) can be studied simultaneously. Off-line processing is devoted to the graphic treatment and statistical analysis of the data. The effects of a single intravenous administration of dihydralazine are presented in order to show the usefulness of the technique.

Computer analysis of cardiovascular activity in conscious unrestrained hypertensive rats of the Lyon strain.

A new computerized technique was developed for the continuous analysis of intra arterially recorded blood pressure (BP) curve in freely moving rats during long periods of time. For each cardiac cycle 5 parameters were calculated on-line and stored. Off-line processing allowed graphical display and statistical analysis. This technique demonstrated that 21 week-old genetically hypertensive rats from the Lyon strain (LH) exhibited higher and more variable BP and dp/dt max. with a lower heart rate than normotensive (LN) and low blood pressure (LL) controls. LL rats differed from LN by a slightly higher systolic and a lower diastolic BP.

Beat-to-beat stroke volume estimation from aortic pressure waveform in conscious rats: comparison of models.

Several methods for estimating stroke volume (SV) were tested in conscious, freely moving rats in which ascending aortic pressure and cardiac flow were simultaneously (beat-to-beat) recorded. We compared two pulse-contour models to two new statistical models including eight parameters extracted from the pressure waveform in a multiple linear regression. Global as well as individual statistical models gave higher correlation coefficients between estimated and measured SV (model 1, r = 0.97; model 2, r = 0.96) than pulse-contour models (model 1, r = 0.83; model 2, r = 0.91). The latter models as well as statistical model 1 used the pulsatile systolic area and thus could be applied to only 47 +/- 17% of the cardiac beats. In contrast, statistical model 2 used the pressure-increase characteristics and was therefore established for all of the cardiac beats. The global statistical model 2 applied to data sets independent of those used to establish the model gave reliable SV estimates: r = 0.54 +/- 0.07, a small bias between -8% to +10%, and a mean precision of 7%. This work demonstrated the limits of pulse-contour models to estimate SV in conscious, unrestrained rats. A multivariate statistical model using eight parameters easily extracted from the aortic waveform could be applied to all cardiac beats with good precision.

Mechanisms of spontaneous baroreflex impairment in lyon hypertensive rats.

This experiment aimed at 1) comparing the spontaneous baroreflex sensitivity (SBRS) in Lyon genetically hypertensive (LH), normotensive (LN), and low blood pressure (LL) rats and 2) assessing some aspects of the mechanisms of its impairment in LH rats. Baroreflex was studied in control animals after an early chronic converting enzyme inhibition with perindopril and after a 4-wk infusion of ANG II in perindopril-treated rats. The SBRS was determined with a previously validated method, using statistical dependence between blood pressure (BP) and heart rate values recorded in freely moving animals. LH rats exhibited high BP, cardiac hypertrophy, and decreased SBRS (LH, 1.3 +/- 0.2; LN, 2.5 +/- 0.4; LL, 2.2 +/- 0.4 beats . min-1 . mmHg-1). Perindopril prevented the development of hypertension and cardiac hypertrophy and normalized SBRS. BP rose in LH and LL rats after ANG II infusion, but only LH rats, which developed a cardiac hypertrophy, had an impaired SBRS (LH, 1.1 +/- 0.2; LN, 2.5 +/- 0.2; LL, 2.8 +/- 0.3 beats . min-1 . mmHg-1). This impairment was partially reversed by an acute ANG II blockade with losartan. These results demonstrate that high BP does not account for the decreased SBRS in LH rats. SBRS impairment could result either from cardiac hypertrophy or from the direct effect of ANG II on the baroreflex loop.

Autonomic nervous system and cardiovascular variability in rats: a spectral analysis approach.

Mechanisms underlying systolic (SBP) and diastolic (DBP) blood pressure and heart rate (HR) beat-to-beat variability were investigated using spectral analysis in conscious genetically normotensive (LN) and hypertensive (LH) adult rats from the Lyon strains. In LN rats, basal SBP, DBP, and HR spectra exhibited peaks in low (LF: 0.38-0.45 Hz) and high (HF: 1.04-1.13 Hz) frequencies. The LF peak of SBP, and even more of DBP, could be attributed to the influence of the sympathetic nervous system as it disappeared after destruction of the sympathetic nerves or a combined alpha- and beta-adrenoceptor blockade, whereas it was higher after blockade of the parasympathetic system. The HF peak of HR, linked to the respiratory rate, was abolished by the parasympathetic system blockade, whereas those of SBP and DBP were enhanced. In LH rats, which exhibit a lower sympathetic activity, the LF peaks of SBP and DBP were less distinct compared with LN controls. We conclude that the LF peak of DBP and the HF peak of HR are likely to represent useful estimates of the sympathetic vascular control and of the parasympathetic cardiac control, respectively.

A new non-invasive statistical method to assess the spontaneous cardiac baroreflex in humans.

1. A new method was developed to evaluate cardiac baroreflex sensitivity. The association of a high systolic blood pressure with a low heart rate or the converse is considered to be under the influence of cardiac baroreflex activity. This method is based on the determination of the statistical dependence between systolic blood pressure and heart rate values obtained non-invasively by a Finapres device. Our computerized analysis selects the associations with the highest statistical dependence. A 'Z-coefficient' quantifies the strength of the statistical dependence. The slope of the linear regression, computed on these selected associations, is used to estimate baroreflex sensitivity. 2. The present study was carried out in 11 healthy resting male subjects. The results obtained by the 'Z-coefficient' method were compared with those obtained by cross-spectrum analysis, which has already been validated in humans. Furthermore, the reproducibility of both methods was checked after 1 week. 3. The results obtained by the two methods were significantly correlated (r = 0.78 for the first and r = 0.76 for the second experiment, P < 0.01). When repeated after 1 week, the average results were not significantly different. Considering individual results, test-retest correlation coefficients were higher with the Z-analysis (r = 0.79, P < 0.01) than with the cross-spectrum analysis (r = 0.61, P < 0.05). 4. In conclusion, as the Z-method gives results similar to but more reproducible than the cross-spectrum method, it might be a powerful and reliable tool to assess baroreflex sensitivity in humans.