Natale R. Musso

Ambulatory Blood Pressure Monitoring: How Reproducible Is It?

We tested the reproducibility of ambulatory blood pressure monitoring (ABPM) by the use of agreement plots. Thirty-two normotensive volunteers underwent ABPM on four separate days (interval 28 days), on the same typical weekday. Sleeping time was restricted to the ABPM nighttime subperiod from 11:00 PM to 7:00 AM. Twenty-four-hour average values-both systolic and diastolic-daytime average values, and nighttime average values, as well as standard deviation (SD) values, were analyzed for differences (analysis of variance). Adaptation occurred from the first to the fourth ABPM, ie, average 24 h, daytime, and nighttime values were lower (-1 to -3 mm Hg) during the fourth recording than the first (P < .05 to P < .01). The agreement analysis showed a surprisingly high agreement among the four data sets (ie, differences from +/-2.54 to +/-5.92 mm Hg; +/-2 SD of the distribution). We concluded that reproducibility of ABPM seems excellent, but adaptation may occur, even in normotensive volunteers under research conditions. Caution must be paid before labeling a patient as hypertensive, because initial ABPM may yield higher values than later monitorings.

L-TYROSINE AND NICOTINE INDUCE SYNTHESIS OF L-DOPA AND NOREPINEPHRINE IN HUMAN LYMPHOCYTES

Catecholamines (CA) were studied in peripheral human lymphocytes in basal conditions as well as after L-tyrosine and/or acetylcholine (ACh) stimulation. Nicotinic and muscarinic receptor activation and blockade were assessed. CA were determined after ultrasonic cell disruption in peripheral lymphocytes after incubation (1 h at 37 degrees C) with the chemicals employed. L-tyrosine significantly increased (P < 0.01) L-Dopa and norepinephrine (NE) content of lymphocytes. ACh in the low microM range did not modify, whereas ACh (60 microM) and (120 microM) significantly increased (P < 0.01), both L-Dopa and NE intracellular levels. L-tyrosine plus ACh (60 microM) or (120 microM) significantly increased (P < 0.01) intracellular L-Dopa and NE versus control, versus L-tyrosine alone and versus ACh alone. The increase was higher than the algebraic sum of the individual increases. Nicotine (250 microM), but not muscarine (50 microM), significantly increased L-Dopa and NE in lymphocytes. Tetraethylammonium (500 microM) (nicotinic blocker), but not atropine (100 microM) (muscarinic blocker), inhibited the ACh-mediated increase of intracellular L-Dopa and NE. These data show that lymphocyte synthesis of CA is under nicotinic control. Since intracellular L-Dopa after L-tyrosine plus ACh increased 6-fold versus basal, 2-fold versus L-tyrosine alone and 3-fold versus ACh alone, it is concluded that ACh might regulate CA synthesis in lymphocytes through an activation of the rate limiting enzyme tyrosine hydroxylase.

Simultaneous Measurement of Plasma Catecholamine (Norepinephrine, Epinephrine, and Dopamine) and Free N—Methyl Dopamine (Epinine) Levels, by HPLC with Electrochemical Detection

Abstract Epinine is the active moiety of ibopamine, a cardiovascular prodrug used in congestive heart failure. This catecholic compound shows dopaminergic and adrenergic properties. Moreover the drug seems to affect plasma catecholamine levels in patients with heart failure. Here we present a method developed for the simultaneous determination of epinine and catecholamine plasma levels. Free epinine and catecholamines were extracted from human venous plasma via an alumina adsorption procedure. The extracts underwent an ion-pair reversed-phase HPLC separation with three-electrode coulometry. Quantitation was made by an internal standard method. Coefficients of variation were 0.997). The detection limits were ≤ 5 pg of each catechol after extraction. This method allows about 50 low cost determination to be done in a working day.

Effects of intravenous epinine administration on left ventricular systolic performance, coronary hemodynamics, and circulating catecholamines in patients with heart failure.

Summary Twenty-six patients with mild to moderate heart failure were studied to determine the effects of epinine infusion (at a rate producing plasma levels similar to those measured after oral administration of 100 mg of the prodrug ibopamine) on left ventricular (LV) function (14 patients), and coronary flow and circulating catecholamines (12 patients). The only significant hemodynamic change at an infusion rate of 0.5 μg/kg/min was a 9% (p < 0.05) decrease in systemic vascular resistance (SVR). At an infusion rate of 1 μg/kg/min (mean free epinine plasma levels 14.3 ± 3.7 ng/ml), heart rate (HR), dP/dtmax (1,405 ± 255 to 1,490 ± 320 mm Hg, NS), (dP/dt)/DP40, and the relaxation rate remained unchanged, but the ejection fraction (EF) increased from 32 to 38% (p < 0.001), cardiac output (CO) increased, and SVR decreased by 22% (p < 0.05). In a separate group of 12 patients, epinine infusion at rates of 0.5–1 μg/kg/min produced no significant changes in coronary blood flow or myocardial oxygen uptake. At these infusion rates, arterial norepinephrine (NE) and dopamine (DA) levels decreased slightly and arterial and coronary sinus epinephrine increased. In conclusion, epinine, at concentrations similar to those achieved during therapeutic use of ibopamine, had negligible effect on myocardial contractility and relaxation rate in heart failure patients. Cardiac pump function was improved by a decrease in SVR rather than by inotropic stimulation. The data also suggest that these low concentrations of epinine may modulate the sympathetic nervous system, but further studies are needed to determine whether this effect could have clinical significance.

Yohimbine effects on blood pressure and plasma catecholamines in human hypertension

The purpose of this study has been to test the hypothesis of an alpha 2-adrenoreceptor alteration in human essential hypertension. The design of the study involved the oral administration of 10 mg yohimbine, an alpha 2-adrenergic antagonist, to 25 healthy volunteers and 29 sex- and age-matched untreated hypertensive patients. Volunteers and patients were studied twice in random order, after placebo or yohimbine treatment, in supine and upright positions. Arterial pressure and heart rate were monitored by servoplethysmomanometry, and venous plasma catecholamines were determined by HPLC with electrochemical detection. Yohimbine induced a significant increase in diastolic pressure only in the hypertensive patients. Plasma norepinephrine was increased significantly in both yohimbine-treated groups, but the percent increase of plasma norepinephrine after the standing test was decreased significantly only in the healthy yohimbine-treated subjects. Plasma dopamine was increased significantly only in the healthy yohimbine-treated subjects. The response of plasma dopamine to the upright position was modified only in the healthy yohimbine-treated subjects. The decrease observed after 2 min of standing was abolished, showing the involvement of alpha 2-adrenoreceptors in the physiologic response of plasma catecholamines in healthy volunteers. Our data may be consistent with some in vivo evidence of an alpha 2-adrenoreceptor desensitization or an alteration in the balance of alpha-adrenoreceptors in human hypertension.

Morphine-Induced TSH Release in Normal and Hypothyroid Subjects

The effects of morphine (10 mg i.v.), an opioid agonist, and of naloxone (10 mg i.v.), an opioid antagonist, on serum levels of TSH and PRL were studied in 7 hypothyroid patients and in 5 normal volunteers. Morphine administration induced a prompt, significant increase in serum TSH and PRL in all subjects. The degree of PRL release after morphine was similar in the two groups, while, as regards TSH, the increase was more evident in hypothyroid subjects. Pretreatment with naloxone (4 mg i.v. 5 min before morphine administration) blocked these effects in all subjects. In contrast, naloxone alone was not able to affect significantly TSH and PRL secretion. Moreover, in 5 other euthyroid volunteers, morphine significantly enhanced the response of TSH and PRL to TRH stimulation (200 micrograms i.v.). These data demonstrate that morphine exerts a stimulatory action on TSH and PRL secretion: the possible mode of action of this drug and the physiologic significance of these findings are discussed.

Blood Pressure Evaluation by Noninvasive and Traditional Methods: Consistencies and Discrepancies Among Photoplethysmomanometry, Office Sphygmomanometry, and Ambulatory Monitoring. Effects of Blood Pressure Measurement

Three hundred eight outpatients referred for hypertension were studied. A continuous beat-to-beat noninvasive recording (Finapres) of blood pressure evaluated the blood pressure increase (9 mm Hg systolic and 4 mm Hg diastolic) induced by office sphygmomanometry. Thereafter, patients underwent a 24-h ambulatory blood pressure monitoring. The evaluation against Finapres showed that office sphygmomanometry overestimates the systolic blood pressure by 3 +/- 36 mm Hg (mean +/- 2 SD) and the diastolic blood pressure by 15 +/- 25 mm Hg (mean +/- 2 SD). Blood pressure monitoring showed similar discrepancies. On the basis of both monitoring (normalcy defined from a population of 550 normotensive subjects) and office sphygmomanometry, patients were considered normotensive, hypertensive (either untreated or under active drug treatment), white coat hypertensive (monitoring below the 95th percentile and sphygmomanometry more than 140/90 mm Hg, either untreated or under active drug treatment), and reverse white coat patients (monitoring over the 95th percentile and sphygmomanometry less than 140/90 mm Hg). Patients showed different levels of alert reaction (the highest in white coat hypertensive and the lowest in reverse white coat hypertensive patients), and a similar increase in blood pressure induced by conventional sphygmomanometry. During initial readings of ambulatory monitoring, blood pressure decreased from the first reading to the third reading. This decrease is related to the increase of blood pressure under sphygmomanometry. Caution should be paid in interpreting results of sphygmomanometry (error level in the single patient as high as +/- 40 mm Hg), and interpreting and averaging results of the first hour of blood pressure monitoring (variably affected by the alert reaction to the clinical environment).

HPLC with electrochemical detection of catecholamines in human plasma. A mini-review

Abstract Since 1970s HPLC received growing attention among catecholamine (CA) assay methods. Earlier methods such as colorimetric (1) or fluorimetric (2) nowadays seem obsolete. Radiotracer methods are usually effective for the determination of norepi-nephrine (NE) and epinephrine (Epi) in human plasma, although the high cost per sample, the poor sensitivity regarding plasma dopamine (DA), the involvement of radiolabelled compounds and the te-

ACETYLCHOLINE-INDUCED, CALCIUM-DEPENDENT NOREPINEPHRINE OUTFLOW FROM PERIPHERAL HUMAN LYMPHOCYTES

Catecholamines (CA) were studied in peripheral human lymphocytes, as well as in the supernatants, after incubation with L-tyrosine and L-dihydroxyphenylalanine (L-Dopa) for 1 h. The effect that the addition of acetylcholine (ACh), Veratridine, lonomycin or KCI had on the outflow of norepinephrine (NE) from lymphocytes was also studied. The effect of the addition of methoxyverapamil (D600, a Ca2+ channel blocker) and cholinergic antagonists had on the ACh-induced NE outflow was assessed. CA were determined by HPLC-ECD, both in the supernatant and in the cell lysates. L-Tyrosine and L-Dopa significantly (P < 0.01) increased intracellular NE. Neither L-tyrosine, L-Dopa, nor vehicle induced a detectable outflow of NE to the supernatants. ACh [120 microM], Veratridine [100 microM], Ionomycin [10 microM] and KCl [50 mM] (with or without the simultaneous addition of L-tyrosine or L-Dopa) all induced a detectable outflow of NE to the supernatant when added 5 min before the end of incubation. NE was not detectable in the supernatant when the chemicals were added 10 to 20 min before the end of the incubation. When the chemicals were added at lower concentrations, erratic secretion or no secretion whatsoever was observed. D600 [100 microM] was able to significantly (P < 0.01) reduce the ACh-induced NE outflow. Tetraethylammonium (nicotinic antagonist), but not atropine (muscarinic antagonist), significantly (P < 0.001) decreased the ACh-induced NE outflow. The outflow of NE from peripheral human lymphocytes was seen. NE secretion seems to be ACh- and calcium-dependent since Veratridine, Ionomycin and KCl are able to induce Ca2+ entry by means of various mechanisms. The Ca2+ channel blocker employed in this study (D600) reduced the ACh-dependent NE outflow. We can conclude that both ACh (through nicotinic receptors) and calcium are involved in the outflow of NE from peripheral human lymphocytes.

Plasma dopamine response to sympathetic activation in man: A biphasic pattern

We studied the plasma catecholamine response to standing and bicycle ergometric tests in 16 normal male subjects. During the standing test (performed in 10 subjects), we observed an early increase in plasma dopamine together with the fast increase in norepinephrine values; in the second half of this test (i.e. from 5 to 10 min of standing), we observed an increase in plasma dopamine levels. During the ergometric test (performed in 6 subjects), we observed a plasma dopamine increase at the maximal exercise; this persisted during the early recumbent recovery phase (6 min), despite the clear-cut decrease of both norepinephrine and epinephrine plasma levels. Our data are not in agreement with previous papers describing a simple increase in plasma dopamine after stimulation. This paper provides no informations regarding the mechanisms of this response of plasma dopamine. Other approaches must be used to study this aspect more directly.