Determination Of Plasma Catecholamines Research Articles

Determination of Catecholamines in a Small Volume (25μL) of Plasma from Conscious Mouse Tail Vein.

The determination of plasma catecholamine levels is commonly used as a measure of the sympathetic nervous system's response to stress and is highly important for diagnosis, therapy, and prognosis of cardiovascular diseases, catecholamine-secreting tumors arising from the chromaffin cells of the sympathoadrenal system, and affective disorders. Diseases in which catecholamines are significantly elevated include pheochromocytoma, Parkinson's disease, Alzheimer's disease, neuroblastoma, ganglioneuroblastoma, von Hippel-Lindau disease, baroreflex failure, chemodectina (nonchromaffin paraganglioma), and multiple endocrine neoplasia. Plasma norepinephrine levels provide a guide to prognosis in patients with stable, chronic, and congestive heart diseases. The method described here for the determination of plasma catecholamines is based on the principle that plasma catecholamines are selectively adsorbed on acid-washed alumina at pH 8.7 and then eluted at a pH between 1.0 and 2.0. Upon injection, catecholamines in elutes were separated by a reversed phase C-18 column. After separation, the catecholamines present within the mobile phase enter the electrochemical detector. Electrochemical detection occurs because electroactive compounds oxidize at a certain potential and thereby liberate electrons that create measurable current. Catecholamines readily form quinones under these conditions, get oxidized, release two electrons, and create current. The electrochemical detector detects this electrical current that linearly correlates to the catecholamine concentration loaded into the ultra-performance liquid chromatography instrument. A 15-min mixing time during the adsorption and desorption steps was found to be optimal. If the washing step was omitted, the catecholamines could not be eluted from the acid-washed alumina. To prevent dilution, the alumina had to be centrifuged and not aspirated to dryness after the washing step. We report here that by changing the range in the electrochemical detector, plasma catecholamines were measured with only 12.5μL of plasma and more reliably with 25μL of plasma. The detection limit was 1ng/mL. This assay method is very useful as blood can be collected from the tail vein in a conscious mouse and the same mouse can be used for time-dependent or age-dependent studies.

Brain fog in neuropathic postural tachycardia syndrome may be associated with autonomic hyperarousal and improves after water drinking.

BackgroundBrain fog is a common and highly disturbing symptom for patients with neuropathic postural tachycardia syndrome (POTS). Cognitive deficits have been measured exclusively in the upright body position and mainly comprised impairments of higher cognitive functions. The cause of brain fog is still unclear today. This study aimed to investigate whether increased autonomic activation might be an underlying mechanism for the occurrence of brain fog in neuropathic POTS. We therefore investigated cognitive function in patients with neuropathic POTS and a healthy control group depending on body position and in relation to catecholamine release as a sensitive indicator of acute stress. The second aim was to test the effect of water intake on cardiovascular regulation, orthostatic symptoms, cognitive function and catecholamine release.MethodsThirteen patients with neuropathic POTS and 15 healthy control subjects were included. All participants completed a total of four rounds of cognitive testing: two before and two after the intake of 500 ml still water, each first in the supine position and then during head-up tilt. At the end of each cognitive test, a blood sample was collected for determination of plasma catecholamines. After each head-up tilt phase participants were asked to rate their current symptoms on a visual analogue scale.ResultsWorking memory performance in the upright body position was impaired in patients, which was associated with self-reported symptom severity. Patients had elevated plasma norepinephrine independent of body position and water intake that increased excessively in the upright body position. The excessive increase of plasma norepinephrine was related to heart rate and symptom severity. Water intake in patients decreased norepinephrine concentrations and heart rate, and improved symptoms as well as cognitive performance.ConclusionBrain fog and symptom severity in neuropathic POTS are paralleled by an excessive norepinephrine secretion. Bolus water drinking down-regulates norepinephrine secretion and improves general symptom severity including brain fog.

A 3-min UPLC-MS/MS method for the simultaneous determination of plasma catecholamines and their metabolites: Method verification and diagnostic efficiency

ObjectiveTo verify a rapid and sensitive ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the quantification of catecholamines and their metabolites, and to validate its efficiency for the diagnosis of phaeochromocytomas and paragangliomas (PPGLs). MethodsPlasma samples were pretreated with solid-phase extraction, followed by a 3-min UPLC-MS/MS analysis to quantify epinephrine (E), norepinephrine (NE), dopamine (DA), metanephrine (MN), normetanephrine (NMN) and 3-methoxytyramine (3-MT), simultaneously. The UPLC-MS/MS method was comprehensively verified and its diagnostic efficiency on PPGLs was tested using 7 PPGLs and 408 non-PPGLs patient plasma samples. ResultsUsing the developed method, the limit of detections (LODs) of the 6 analytes ranged from 0.0002 nmol/L (MN) to 0.0250 nmol/L (NE), while the lower limit of measuring intervals (LLMIs) ranged from 0.05 nmol/L (E, MN and NMN) to 0.10 nmol/L (NE and DA). The reportable ranges were 0.05–30.00 nmol/L for E, MN and NMN, 0.10–30.00 nmol/L for NE and DA, 1.00–300.00 pg/mL for 3-MT. No significant matrix effect was detected after correcting using internal standard. Besides, intra-day and inter-day precision were also within acceptance criteria with coefficient of variations (CVs) ≤ 15% and recoveries ranged from 95% to 115% for all the 6 analytes. The carryover effect was lower than 10%. Its diagnostic efficiency for PPGLs was significantly increased, the areas under the receiver operating characteristic (ROC) curves were increased from 68.7% to 89.1% (using E, NE and DA) to 75.2%–99.9% (using MN, NMN and 3-MT). ConclusionThis study verified a rapid UPLC-MS/MS method for the determination of catecholamines and their metabolites in human plasma. It showed high diagnostic efficiency and will serve as an important tool to avoid the risk for missing patients with PPGLs.

LC-MS/MS determination of plasma catecholamines after selective extraction by borated zirconia.

For the first time, boronate affinity chromatography and metal oxide affinity chromatography mechanisms for cis-diol-compounds extraction are simultaneously realized on a single material, termed borated zirconia. This material was prepared by hydrolyzing zirconium butoxide with boric acid under non-aqueous environment. The diameter of formed particles is around 200nm. By extracting catechol under different pH conditions or with phosphate ion competition, the dual affinity mechanisms of borated zirconia are confirmed. Benefiting from such unique feature, borated zirconia can function well under neutral condition. By using borated zirconia for dispersive solid phase extraction, specific capture of cis-diol-containing catecholamines, including epinephrine (E), norepinephrine (NE) and dopamine (DA), was achieved. A reliable LC-MS/MS method was established and validated for quantification of these target analytes in plasma samples after derivatisation with benzoyl chloride. The linear ranges are 0.010-0.200ng·mL-1 for E and DA, and 0.050-1.000ng·mL-1 for NE. The limits of quantification are 0.008, 0.020 and 0.004ng·mL-1 for E, NE and DA respectively. By analyzing samples from healthy volunteers and schizophrenia patients, the plasma concentrations of E, NE and DA were found to behigher for the latter. Graphical AbstractSchematic representation of boronate combined metal oxide affinity chromatography (BMOAC) extraction of cis-diol catecholamines from human plasma by borated zirconia following with benzoyl chloride derivatization and LC-MS determination.

Fast determination of catecholamines in human plasma using carboxyl-functionalized magnetic-carbon nanotube molecularly imprinted polymer followed by liquid chromatography-tandem quadrupole mass spectrometry

A novel, simple and sensitive method based on the use of dispersive micro-solid-phase extraction (d-μ-SPE) procedure combined with ultra-fast liquid chromatography–tandem quadrupole mass spectrometry (UFLC–MS/MS) for the determination of catecholamines, i.e., dopamine (DA), norepinephrine(NE) and epinephrine (E), was developed and validated. The novel catecholamines molecularly imprinted polymer (MIP) on the surface of carboxyl-functionalized magnetic-carbon nanotube (CF@m-CNTs-MIP) was synthesized and characterized by vibrating sample magnetometer (VSM), scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS). The CF@m-CNTs-MIP was used as the d-μ-SPE sorbent to extract catecholamines from human plasma samples. The obtained results demonstrated the higher extraction capacity of CF@m-CNTs-MIP with recoveries between 87.5–110%. The limits of quantification (LOQs) for NE, E and DA were 76ng/L, 18ng/L and 10ng/L, respectively. Validation results on linearity, specificity, accuracy, precision and stability, as well as on application to the analysis of catecholamines in 120 healthy volunteers demonstrated the applicability to clinical studies.

Influence of aerobic training on neurohormonal and hemodynamic responses to head-up tilt test and on autonomic nervous activity at rest and after exercise in patients after bypass surgery

Little is known about the influence of aerobic training on the neurohormonal and hemodynamic responses to head-up tilt (HUT) and on autonomic balance at rest and after exercise in optimally treated, low risk post-coronary artery bypass grafting (CABG) patients. One hundred male patients, mean age 56 ± 6 years, 3 months after CABG, were randomized to either 6-week training on cycloergometer, 3 times a week, at 70-80% of max tolerated heart rate (HR) (training group, n = 50) or to a control group (n = 50). At baseline and at the end of the study, all patients underwent: (1) cardiopulmonary exercise test with HR recovery (HRR) assessment; (2) 60% HUT during which HR, blood pressure (BP), stroke volume (SV by impedance cardiography) were monitored and blood samples were taken for determination of plasma catecholamines and ANP levels, and plasma renin activity; (3) assessment of HR variability (HRV) in the time and frequency domains at rest. During the final tests, HUT-induced changes in HR, BP, SV, cardiac output, total peripheral resistance, and noradrenaline were significantly lower in training group than in controls. In addition, after training faster post-exercise HRR, increased SDNN and a tendency towards an increase in the high frequency HRV power spectrum were found. Aerobic training improved neurohormonal and hemodynamic responses to head-up tilt test and favorably modified sympatho-vagal balance in low risk post-CABG patients.

I005 Chemical denervation of sympathetic nervous system induces abnormal myocardial architecture

The role of autonomic nervous system (ANS) on heart function modulation is well-known. By contrast, ANS role on myocardial tissue architecture has scarcely been investigated. The aim of the present work was to investigate changes in heart tissue architecture after chemical sympathetic denervation by 6OH-Dopamine (6OH-DA) in mice. Two months old mice (n=18) received 3 injections of 6OHDA (200 mg/kg, ip) or saline (n = 6) at 3 days of interval. At 15 and 30 days after first injection, ECG was recorded (PowerLab, DSI) under anaesthesia and heart rate spectral variability (HRV) was performed (FFT) in low frequency (LF: 0.15-1.5 Hz) and high frequency (HF: 1.5-5 Hz) ranges; LH/HF ratio was also calculated. After sacrifice, blood was withdrawn for plasma catecholamine determination (HPLC). Heart tissue was fixed (formaldehyde 10 %) for histology or frozen for western blot analysis (tyrosine hydroxylase, TH). When compared to controls (1410±145 pg/ml) plasma norepinephrine levels were significantly lower at D15 (766±186 pg/ml) and D30 (675±288 pg/ml) after 6OH-DA without significant change in epinephrine levels. TH expression was absent at D15 and present but significantly lower than in controls at D30. When compared to controls (48.5±6.2 %), LF HRV was significantly reduced at D15 (31.6±5.4 %) but not at D30 (58.2±16.2 %) without any change in HF. LF/HF ratio was lower in 6OH-DA treated mice at D15 (0.49±0.13 vs 1.29±0.17 in controls) but was normal at D30 (1.63±0.31). At D15, hearts from 6-OH-DA treated mice exhibited mild structural abnormalities with wavy cardiomyocyte appearance in septum. At D30, histological abnormalities concerned whole myocardium with myocytes intersecting at various angles with bundles wavy appearance. Variability in cell size with anisocaryosis, attenuated myocytes with perinuclear halo and shaped nuclei were observed. No inflammation, interstitial fibrosis or necrosis were noticed. This study suggests that heart denervation induces myocardial tissue disorganization. Relationship between these pathological changes and sympathetic nerve destruction and/ or catecholamine depletion remains to be elucidated. Apart from physiological significance, these results also bring new structural basis to explain increased risk of cardiac disease during human autonomic failure.

Study on the correlation of the autonomic nervous system responses to a stressor of high discomfort with personality traits

The present study investigated Eysenck's predictions concerning the correlation of personality to arousal at higher levels of stress. Twenty young adults were exposed to a physical stress causing great discomfort, specifically a cold wind (4 °C at 60 km/h) exposure to the face for 3 min. Autonomic nervous system (ANS) responses were measured by continuous heart rate and plasma catecholamine determinations before, during and after the test. At the end of the test, the participants gave a rating of discomfort on a 0 to 10 scale. The personality traits were assessed with the Big-Five Inventory test (BFI). Results indicated that higher levels of trait extraversion were positively correlated with discomfort ratings and with the increased heart rate and the noradrenaline responses. Neuroticism was negatively correlated to discomfort and the autonomic responses. These findings tend to support Eysenck's theory on the role of personality on arousal at higher levels of stress. It is also proposed that the better tolerance to this severe stress observed with neuroticism is correlated to a certain habituation process caused by light to moderate arousal frequently experienced by participants with this personality trait during their daily activities.

Haemodynamic changes early in prodromal symptoms of vasovagal syncope.

Vasovagal syncope (VVS) is often preceded by prodromal symptoms. The haemodynamic changes occurring during the prodrome have not been systematically investigated. The aim of the present study was to investigate the behaviour of blood pressure (BP), heart rate (HR) and sympathetic activity at the beginning of the prodrome in patients with tilt-induced VVS. Sixty-three patients with VVS underwent tilt testing. BP and HR were measured and blood samples for plasma catecholamine determination were obtained during the test. Twenty-seven patients developed syncope of whom all had a prodrome. From the last scheduled measurement before prodromal symptoms to the beginning of the prodrome, both systolic and diastolic BP decreased in all patients (from 105 +/- 16 to 74 +/- 20 mmHg, P<0.001, and from 68 +/- 13 to 51 +/- 12 mmHg, P<0 001, respectively) and HR decreased in 18 (67%) (from 89 +/- 22 to 80 +/- 25 beats/ min P<0 02). At the onset of loss of consciousness both BP and HR showed a further decrease (P<0.001). Plasma adrenaline significantly increased from the last sample before prodromal symptoms to the beginning of the prodrome (P<0.01) and showed a further increase during loss of consciousness (P<0.05), whereas plasma noradrenaline did not increase, as an expression of inhibition of sympathetic neural outflow. These results demonstrate that in patients with tilt-induced VVS, BP is consistently decreased at the beginning of prodromal symptoms because of the withdrawal of sympathetic activity, and HR is often reduced, probably because of increased vagal activity. We may infer that similar haemodynamic features also occur during spontaneous VVS.

Pheochromocytoma in multiple endocrine neoplasia type 2: a prospective study.

The aim of this prospective study is to update our knowledge of the chronology of pheochromocytoma occurrence in multiple endocrine neoplasia type 2 (MEN 2), and to better manage MEN 2 patients after the genetic diagnosis. Eighty-seven non-index gene carrier MEN 2 patients were included in this prospective study: 84 patients with MEN 2A (from 52 families) and 3 with MEN 2B (from 3 families). Medullary thyroid carcinoma (MTC) was diagnosed by measuring plasma calcitonin in basal conditions or after pentagastrin stimulation. The search for pheochromocytoma consisted of clinical evaluation, 24 h determination of urinary catecholamines and adrenal imaging. The mean age at genetic diagnosis of MEN 2 was 14.0+/-7.0 years, the mean duration for the follow-up was 7.6+/-2.8 years. All 87 patients had a MTC detected at the same time as the genetic diagnosis was made. Urinary catecholamine measurements led to the diagnosis of pheochromocytoma and a combination of imaging techniques enabled the correct localization of both unilateral or bilateral adrenal involvement. Pheochromocytoma was detected simultaneously with MTC in only seven patients, and seven others were detected throughout the follow-up. Of the 14 patients with pheochromocytoma, 11 had bilateral involvement: nine were initially bilateral and two became so during follow-up. This study demonstrates that in MEN 2, MTC is the lesion which appears earliest. Pheochromocytoma develops later during the evolution of the disease, and necessitates regular clinical and biological monitoring throughout follow-up. Determination of urinary and/or plasma catecholamines and metanephrines should be performed to detect pheochromocytoma. Imaging techniques lead to the detection of both unilateral and bilateral pheochromocytoma, thus making video-assisted laparoscopic adrenalectomy possible.

Preoperative clonidine attenuates stress response during emergence from anesthesia

Study Objectives: To investigate whether a single preoperative IV dose of clonidine blunts the hemodynamic and hyperadrenergic responses not only to intubation, but also to extubation. Design: Randomized, double-blind, placebo-controlled study Patients: 29 ASA physical status I and II patients (ages 18–65) who were scheduled for noncardiac, elective surgery. Patients were randomly assigned to either receive clonidine 3 μg/kg IV immediately before anesthesia induction or placebo. Interventions: Insertion of a 14 G cannula in a large cubital vein for the determination of plasma catecholamines using local anesthesia. Insertion of a radial artery catheter for measuring blood pressure (BP) using local anesthesia. Transthoracic echocardiography to determine cardiac output (CO). Measurements: Heart rate (HR), mean arterial pressure (MAP), CO, and plasma catecholamine concentrations. Measurements were performed: before induction (baseline), during intubation, 10 min after intubation, after surgery, during extubation, and 10 min after extubation. Main Results: During intubation MAP, HR, and CO were lower in the clonidine group. Compared with baseline measurements, MAP and CO increased less in the clonidine group during intubation. During extubation, MAP was lower in the clonidine group. CO and MAP increased less as compared with baseline measurements in the clonidine group. Compared with the measurements after surgery CO less in the clonidine group during extubation ( p < 0.05 for all results). Conclusions: A single preoperative IV dose of clonidine (3 μg/kg) blunts the hemodynamic responses due to extubation in noncardiac surgery of intermediate duration.

Pheochromocytoma with Normal Urinary and Plasma Catecholamines but Elevated Plasma Free Metanephrines in a Patient with Adrenal Incidentaloma

We present an asymptomatic 75-year-old patient (normal renal function, no hypertension) with adrenal incidentaloma (5 cm) who repetitively had had negative screening tests for pheochromocytoma by determination of urinary and plasma catecholamines. The attempt then to exclude malignant disease by fin

Automatic semi-microcolumn liquid chromatographic determination of catecholamines in rat plasma utilizing peroxyoxalate chemiluminescence reaction.

A fully automated and highly sensitive method with a semi-microcolumn liquid chromatography system for the determination of rat plasma catecholamines (CAs) was developed. Automated on-line extraction of CAs in diluted plasma using a precolumn packed with strong acidic cation exchange resin was coupled with separation of CAs on a semi-microcolumn (250 x 1.5 mm id). fluorogenic derivatization with ethylenediamine and finally postcolumn peroxyoxalate chemiluminescence detection utilizing bis[2-(3,6,9-trioxadecanyloxycarbonyl)-4-nitrophenyl]oxalate (TDPO) and hydrogen peroxide. The detection limits were 0.91, 0.36 and 1.1 fmol for norepinephrine (noradrenaline), epinephrine (adrenaline) and dopamine, respectively, at a signal-to-noise ratio of 3. A good linearity of the calibration curve for each CA was observed in the range of 5.0 to 500 fmol for each CA using N-methyldopamine (N-MeDA) as an internal standard. The RSD for the proposed method (n = 5) were 3.7-9.5% for the intra-day assay and 6.6-10.0% for the inter-day assay. The volume of rat plasma required for the determination of CAs was 10 microliters.

Chemiluminescence determination of catecholamines in human blood plasma using 1,2-bis(3-chlorophenyl)ethylenediamine as pre-column derivatizing reagent for liquid chromatography

A highly sensitive method for the determination of the human blood plasma catecholamines (norepinephrine, epinephrine and dopamine) is described which employs liquid chromatography (LC) with chemiluminescence detection. Solid-phase extraction using cation exchange cartridges was used for selective and quantitative isolation of the catecholamines and isoproterenol (an internal standard) from 20 μl of human plasma containing 0.1 pmol ml −1 isoproterenol. The amines and isoproterenol, derivatized with 1,2-bis(3-chlorophenyl)ethylene diamine, were separated on a reversed-phase LC column with isocratic elution using a mixture of imidazole buffer (120 mM, pH 5.8), methanol, acetonitrile (13 : 4 : 18, v/v/v). The eluate was detected by a post-column chemiluminescence reaction system using bis[4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl]oxalate and hydrogen peroxide. The detection limits (a signal-to-noise ratio = 3) for each catecholamine are 40–120 amol in a 100 μl injection volume.

Determination of catecholamines in human plasma by high-performance liquid chromatography with electrochemical detection

In the present study, assays were improved for the determination of catecholamines in human plasma. High-performance liquid chromatography with electrochemical detection was employed for quantitative analysis. The influence of various parameters on chromatographic performance, such as the composition and the pH of the mobile phase, and the detection potential, was investigated. An accurate solid-phase extraction procedure, after catecholamine complexation with diphenylborate, was developed. The efficiency yield for all catecholamines was in the range 92–98%. Relative standard deviation values for repeatability and for intermediate precision were less than 2% and 3%, respectively, for all three analytes.

Reduced GABA inhibition of sympathetic function in renal-wrapped hypertensive rats.

Animals with bilateral cannulas in the paraventricular nucleus were made hypertensive by a one-kidney, figure eight renal wrap procedure or sham operated. Femoral artery and vein catheters were inserted for arterial pressure measurement and plasma catecholamine determination. After recovery and 4 days after hypertension surgery, bicuculline methiodide or muscimol was microinjected into the paraventricular nucleus. In some rats, nitroprusside was infused intravenously to reflexly stimulate the sympathetic nervous system. In control rats, bicuculline increased blood pressure, heart rate, and plasma norepinephrine and epinephrine concentrations. In contrast, in hypertensive rats blood pressure did not change while the heart rate response was maintained. Plasma norepinephrine and epinephrine responses were reduced 75 and 68%, respectively. Muscimol injections decreased arterial pressure in the hypertensive rats. Heart rate responses to nitroprusside were similar in the two groups of rats, while the plasma catecholamine responses were attenuated in the hypertensive animals. These data suggest that GABA function in the paraventricular nucleus is reduced in renal wrap hypertension.

High-performance liquid chromatographic determination of plasma catecholamines during alpha-methyldopa therapy.

High-performance liquid chromatographic determination of plasma catecholamines during alpha-methyldopa therapy.

Determination of catecholamines in sheep plasma by high-performance liquid chromatography with electrochemical detection: comparison of deoxyepinephrine and 3,4-dihydroxybenzylamine as internal standard

3,4-Dihydroxybenzylamine (DHBA) is commonly used as the internal standard in HPLC catecholamine assays. Sheep are frequently used in studies of cardiovascular physiology and in such studies measurement of catecholamines is important. The recovery of DHBA from sheep plasma is, however variable and poor. Therefore, we have developed a reliable and sensitive HPLC-ED method with alumina extraction for measurement of catecholamines in sheep plasma using deoxyepinephrine (epinine) as the internal standard. Separation was performed on a μBondapak C 18 column (300×3.9 mm, 10 μm) with a mobile phase containing 2% acetonitrile and 98% buffer (0.05% sodium acetate-0.02% EDTA-0.013% sodium heptanesulfonate), pH 3.25. The extraction of epinine from water, human plasma, dog plasma and sheep plasma did not differ ( p>0.05), but extraction of DHBA from sheep plasma was significantly impaired ( p<0.0001). The R 2 of regression curves ( n=5) of norepinephrine (NE) (25.02 pg/ml-1.00 ng/ml) and epinephrine (E) (25.82 pg/ml-1.03 ng/ml), using epinine as internal standard were greater than 0.99. The intra- and inter-day coefficients of variation were 2.11–11.15 and 0.88–12.60% for NE and 1.12–10.91 and 2.88–12.60% for E, respectively. The detection limits for NE and E are 12 pg/ml. The technique described has the advantage that it allows the simultaneous determination of both endogeneous and [ 3H]norepinephrine in sheep plasma using a sensitive and reproducible HPLC technique.

Effects of intravenous and oral clonidine on hemodynamic and plasma-catecholamine response due to endotracheal intubation

To investigate the effects of intravenous (IV) versus oral clonidine on alterations of heart rate (HR), mean arterial pressure (MAP), cardiac output (CO), and plasma-catecholamines due to endotracheal intubation. Randomized, double-blind, placebo-controlled study. University hospital surgery operating room. 33 ASA physical status I patients were randomly assigned to either receive clonidine 3 micrograms/kg IV immediately prior to anesthesia induction, clonidine 4 micrograms/kg orally 90 minutes prior to anesthesia induction, or placebo. Insertion of a 14 G cannula in a large cubital vein for the determination of plasma-catecholamines using local anesthesia. Insertion of a radial artery catheter for measuring blood pressure (BP) using local anesthesia. Transthoracic echocardiography determined CO. Heart rate, MAP, CO, and plasma-catecholamine concentrations were measured. Measurements were performed prior to induction, during intubation, and 10 minutes after intubation. During endotracheal intubation, MAP was significantly lower in the IV clonidine group compared with the placebo and the oral clonidine groups. Cardiac output was significantly lower in the IV clonidine group only. In contrast to the placebo group, norepinephrine plasma concentrations did not increase in either clonidine group. Significant alterations of epinephrine plasma concentrations due to intubation were not observed in either group. Hemodynamics after intubation were not impaired by clonidine treatment. In conclusion, IV clonidine reduced stress response to endotracheal intubation compared with placebo. Oral clonidine at the dose used was less effective in blunting hemodynamic stress response than IV clonidine.

Spontaneously hypertensive rat Y chromosome increases indexes of sympathetic nervous system activity.

Previous studies from our laboratory have demonstrated that the Y chromosome from the spontaneously hypertensive rat (SHR) is responsible for a significant portion of the elevated blood pressure and also produces an earlier pubertal rise in plasma testosterone. We performed the following studies to determine whether the SHR Y chromosome raises blood pressure by sympathetic nervous system responses as measured by adrenal chromogranin A and plasma and tissue catecholamines. Male SHR from the University of Akron colony were studied from 5 to 20 weeks of age. Blood pressure was measured by tail-cuff, tail artery cannulation, and aortic telemetry (Data Sciences); acute (air stress) and chronic (territorial colony) social stressors were compared; blood was collected for determination of plasma catecholamines; and adrenal glands were analyzed at 15 weeks for catecholamines. Rats with the SHR Y chromosome had higher blood pressure and plasma norepinephrine than those with the normotensive Wistar-Kyoto (WKY) Y chromosome. However, the SHR Y chromosome did not significantly change responsiveness to acute or chronic stressors. Phentolamine and clonidine prevented the stress responses. Adrenal chromogranin A levels were elevated 37% and 40% and adrenal norepinephrine content 29% and 100% at 4 and 10 weeks of age, respectively, in rats with an SHR Y chromosome compared with WKY. Chemical sympathectomy normalized blood pressure in all strains and significantly reduced norepinephrine (36% to 41%) in all strains except in WKY, which already had a normal blood pressure. In conclusion, the SHR Y chromosome appears to increase the chronic sympathetic nervous system. A potential mechanism could be a Y locus that influences chronic sympathetic nervous system activity, which may reinforce neurohumoral factors and structural components of the vessel wall, accelerating the development of hypertension.