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PubMed2005

Measurement of plasma catecholamines in small samples from mice.

INTRODUCTION: A method is described by which it is possible to obtain measurements of plasma catecholamines in small samples from resting chronically catheterized mice. METHODS: Standard alumina extraction procedures were systematically altered to maximize the recovery of catecholamines from 25-mul samples. The technique used commonly available HPLC with electrochemical detection (EC) equipment which was optimized according to the manufacturers' guidelines. RESULTS: The limit of detection is 40 pg/ml plasma noradrenaline and 20 pg/ml of plasma adrenaline and the resting levels were 300 pg/ml for noradrenaline and 80 pg/ml for adrenaline, both much lower than previously published. Comparison of resting levels from catheterized mice with those obtained by CO2 plus decapitation and rapid decapitation reveal increases comparable to those reported following immobilization stress. The catecholamines adrenaline and noradrenaline were differentially increased by the two euthanasia methods. DISCUSSION: The method was reliable, simple to perform and adequately sensitive. The resting levels of plasma catecholamines in mice are lower than previously published using a different sampling method. Differences in the norepinephine and adrenaline increases produced by the different methods of euthanasia suggest caution in the selection of method in studies of sympathetic function.

Animals↗
PubMed1995

Increase of myocardial inhibitory G-proteins in catecholamine-refractory septic shock or in septic multiorgan failure.

PURPOSE: The aim of the study was to investigate the mechanisms of myocardial catecholamine refractoriness in septic shock. METHODS: The inhibitory guanine nucleotide-binding proteins (Gi alpha) were studied with pertussis toxin labeling and radioimmunologically in myocardium from patients who died while in catecholamine-refractory septic shock and from patients who died of noncardiac disease. RESULTS: An increase by 62% (immunological Gi alpha) and 221% (pertussis toxin substrate) of myocardial Gi alpha was observed in patients with catecholamine-refractory shock compared with controls. The increases of Gi alpha were greater than those found in chronic heart failure reported earlier. CONCLUSIONS: An increase in the expression of Gi alpha could also be important in conditions other than chronic heart failure, eg, septic shock. An increase of Gi alpha could play a pathophysiologically relevant role in catecholamine refractoriness in septic shock and could provide a target for pharmacologic treatment in this condition.

Adult↗
PubMed1998

Ibogaine acts at the nicotinic acetylcholine receptor to inhibit catecholamine release.

In an effort to determine mechanisms of action of the putative anti-addictive agent ibogaine, we have measured its effects on catecholamine release in a model neuronal system, cultured bovine chromaffin cells. Various modes of stimulating catecholamine release were used including nicotinic ACh receptor activation, membrane depolarization with elevated K+ and Na+ channel activation with veratridine. In addition, because ibogaine has been reported to interact with kappa opioid receptors, we tested whether kappa receptor antagonists could reverse ibogaine's effects on catecholamine release. Ibogaine, at low concentration (<10 microM) was found to selectively inhibit nicotinic receptor-mediated catecholamine release, while having no significant effect on release evoked by either veratridine or membrane depolarization with elevated K+. The inhibitory actions of ibogaine and the kappa agonists were not reversed by preincubation with the opioid antagonists nor-binaltorphimine or naltrexone, suggesting that these inhibitory effects are not mediated by the kappa opioid receptor. The effects of low dose (10 microM) ibogaine were rapidly reversible, while the inhibitory effects of higher ibogaine doses persisted for at least 19 h following ibogaine washout. The results provide evidence for a mechanism of action ibogaine at the nicotinic ACh receptor. The results are consistent with a model in which the initial high transient brain concentrations (100 microM) of ibogaine act at multiple cellular sites and then have a selective action at the nicotinic ACh receptor cation channel following its metabolism to lower brain concentrations. The present findings are relevant to potential anti-addictive actions of ibogaine and to the development of drugs to combat nicotine addiction.

Animals↗
PubMed1999

Increased catecholamine levels in specific brain regions of a rat model of depression: normalization by chronic antidepressant treatment.

Alterations in catecholamine levels and neurotransmission have been shown in depressive disorders. However, the exact sites of alterations and the relation between these alterations to the etiology of the disease and the effectiveness of antidepressant therapy are poorly understood. In this study, catecholamine levels and metabolism were measured in specific brain regions of a genetic rat model of depression [Flinders Sensitive Line (FSL) rats], and compared to normal Sprague-Dawley rats. Norepinephrine levels were found to be two to threefold higher in the nucleus accumbens, prefrontal cortex, hippocampus and median raphe nucleus of FSL rats as compared with control Sprague-Dawley rats. Dopamine levels were sixfold higher in the nucleus accumbens and twofold higher in the striatum, hippocampus and hypothalamus of FSL rats as compared with control Sprague-Dawley rats. After chronic treatment with the antidepressant desipramine, the immobility score in a swim test, as a measure of a behavioral deficit, as well as catecholamine levels of the FSL rats became normalized, but these parameters in the control rats did not change. The results indicate that the behavioral deficits expressed in the FSL model for depression correlate with increased catecholamine levels in specific brain sites, and further suggest the FSL rats as a model for elucidation of the molecular mechanism of clinically used antidepressant drugs.

Adrenergic Uptake Inhibitors↗
PubMed1998

Simultaneous measurement of catecholamines and kallikrein in urine using boric acid preservative.

Catecholamines, dopamine and the renal kallikrein-kinin system may participate in the pathogenesis of hypertension. In the past these systems have been studied independently in isolation. Attempts to study the systems together have been hampered by incompatibility of the current urine preservatives for the two assays involved. In order to measure acid-stable catecholamines and acid-labile kallikrein enzyme together, we have established boric acid solution as a preservative by comparing the stability of urinary catecholamines stored in it with the commonly employed preservative, hydrochloric acid (HCl) as well as the stability of urinary kallikrein in untreated urine with and without boric acid at ambient temperatures for 24 and 48 h, and at -20 degrees C for 2 weeks and 1, 2 and 3 months. The stability of other common urine parameters including cortisol, electrolytes, creatinine and protein, was also investigated after storage with boric acid at ambient temperature for 24 h. Our results showed that there was a good agreement between the measurements of urinary catecholamines stored in both HCl and boric acid and that the latter did not interfere with measurements of urinary kallikrein or other common urine parameters.

Albuminuria↗
PubMed1996

Simultaneous inhibition of catechol-O-methyltransferase and monoamine oxidase A: effects on hemodynamics and catecholamine metabolism in healthy volunteers.

OBJECTIVE: To evaluate the effects of simultaneous pharmacologic inhibition of catechol-O-methyltransferase (COMT) and monoamine oxidase type A (MAO-A) on hemodynamics and catecholamine metabolism in healthy volunteers at rest and during exercise. BACKGROUND: Entacapone, a COMT inhibitor, is studied as an adjunct to levodopa treatment in patients with Parkinson's disease. Moclobemide, an MAO-A inhibitor, is already in clinical use as an antidepressant. It is likely that entacapone and moclobemide will be used concomitantly in the future in patients who have both Parkinson's disease and depression. It was therefore considered to be important to investigate the tolerability of combined COMT and MAO-A inhibition with entacapone and moclobemide. DESIGN AND METHODS: This was a randomized, single-dose, double-blind crossover study of 12 healthy male volunteers. The treatments were either placebo, 200 mg entacapone, 150 mg moclobemide, or the combination of entacapone and moclobemide in single doses. Heart rate, blood pressure, impedance cardiography, and plasma concentrations of catecholamines and their metabolites were measured both at rest and during submaximal standardized bicycle exercise. RESULTS: Entacapone and moclobemide (either alone or in combination) did not change heart rate, blood pressure, or any hemodynamic parameter at rest or during exercise compared with placebo. Neither were the concentrations of norepinephrine and epinephrine in plasma influenced. Both drugs had the expected effects on catecholamine metabolite concentrations in plasma. The decrease in the concentration of 3-methoxy-4-hydroxyphenylglycol (MHPG) induced by moclobemide was not potentiated by entacapone. CONCLUSION: The combined use of therapeutic single doses of entacapone and moclobemide in healthy volunteers did not affect the hemodynamics or concentrations of unconjugated norepinephrine and epinephrine in plasma. Other mechanisms are capable of regulating the concentrations of norepinephrine and epinephrine in circulating blood (and apparently also at receptors in the heart and vascular tissue) when both COMT and MAO-A activity are inhibited to a significant extent. This was also the case during marked sympathetic stimulation. The changes in the catecholamine metabolite concentrations provide evidence of effective COMT and MAO inhibition. Concentrations of MHPG in plasma are determined mainly by MAO-A activity because COMT inhibition did not have an additional effect on the moclobemide-induced decrease in plasma MHPG.

Administration, Oral↗
PubMed1998

Properties of ginseng saponin inhibition of catecholamine secretion in bovine adrenal chromaffin cells.

To investigate the relationship between the inhibitory effects of ginseng saponins (ginsenosides) on acetylcholine-evoked secretion of catecholamines and the structures of ginsenosides, we examined the effects of ginsenoside-Rg3 and -Rh2, which are panaxadiol saponins, 20(R)- and 20(S)-ginsenoside-Rg2, which are epimers involving the hydroxyl group at C-20 of sapogenin, and other plant saponins on the acetylcholine-evoked secretion of catecholamines from cultured bovine adrenal chromaffin cells. The ginsenoside-Rg3 (1-100 microM) and -Rh2 (10-100 microM) greatly reduced the acetylcholine-evoked secretion in a concentration-dependent manner comparable to that of ginsenoside-Rg2, a panaxatriol saponin, which was the most potent inhibitor in our previous study. 20(R)- and 20(S)-ginsenoside-Rg2 (1-100 microM) similarly reduced the acetylcholine-evoked secretion. In contrast, saikosaponin-a, glycyrrhizin and the cardiac glycosides (100 nM-100 microM), digitoxin and digoxin, had no significant inhibitory effect on catecholamine secretion. Saikosaponin-c (10-100 microM), however, had an inhibitory effect, which was less than that of ginsenoside-Rg2 and -Rg3. These results strongly suggest that the inhibitory effects of ginsenosides on the acetylcholine-evoked secretion of catecholamines from bovine adrenal chromaffin cells are a unique property of ginseng. Further, the relationship between the inhibitory effects and the structures of ginsenosides is discussed.

Animals↗
PubMed1975

The role of internal spermatic vein plasma catecholamine determinations in subfertile men with varicoceles.

Catecholamine concentrations were determined in left spermatic vein blood of 18 patients with varicocele. These values were compared with concentrations of catecholamines in the right femoral vein. Of 18 patients studied, 6 had moderate to marked elevations of spermatic vein catecholamine concentrations. Norepinephrine was the fractioned component that was elevated. In an average 1-year follow-up the group with elevated catecholamine levels had a much better semen improvement percentage and pregnancy rate than the other group. Chronic exposure of testicular vasculature to norepinephrine may account for the spermatogenic dysfunction encountered in varicocele patients. Once this exposure is interrupted, there may be reversibility of function and the fertility prognosis may improve. The presence of other adrenal or renal metabolites detrimental to spermatogenesis must be considered.

Adult↗
PubMed1987

Analysis of urinary catecholamines by high-performance liquid chromatography in the presence of labetalol metabolites.

Common sample preparation methods for catecholamines lead to contamination with metabolites of labetalol, an anti-hypertensive drug. When the extracts are analyzed by cation-exchange high-performance liquid chromatography, these metabolites are separated from the catecholamines, but their strong retention lengthens the analysis time. A procedure has been developed for complete removal of these drug metabolites from acidified urine by the use of XAD-4 resin. Loss of catecholamines is monitored by an internal standard. This pretreatment can be combined with extraction by weak cation-exchange resin and borate elution to simplify catecholamine analysis for patients receiving labetalol.

Catecholamines↗
PubMed1991

Improvements in automated analysis of catecholamine and related metabolites in biological samples by column-switching high-performance liquid chromatography.

Previously two fully automated methods based on column switching and high-performance liquid chromatography have been described, one for plasma and urinary catecholamines and the other for catecholamine urinary metabolites. Improvements in these methods, after 3 years of routine application, are now reported. The sample processing scheme was changed in order to eliminate memory effects and, in the procedure for plasma catecholamines, a pre-analytical deproteinization step was added which enhances the analytical column lifetime. The applied voltages for the electrochemical detector have been optimized, resulting in an automated method, suitable for the simultaneous determination of vanillylmandelic acid, 3,4-dihydroxyphenylacetic acid, homovanillic acid and 5-hydroxyindoleacetic acid. The sensitivity of the methods allows the detection of 2-3 ng/l of plasma catecholamines and 0.01-0.06 mg/l of urinary metabolites. Also, it is possible to switch from one method to the other in only 30 min. The normal values obtained from 200 healthy people are reported, together with a list of 57 potential interfering substances tested.

Catecholamines↗
PubMed2000

Determination of catecholamines in pheochromocytoma cell (PC-12) culture medium by microdialysis-microbore liquid chromatography.

An in vitro microdialysis system was constructed for the measurement of catecholamines in pheochromocytoma cell culture medium. The novel microdialysis device is composed of a petri dish, a dialysis membrane and two transmission tubes. The dialysis membrane is located in the space of a petri dish such that it is immersed in the culture medium. Catecholamines contained in the culture medium diffused into a designed dialysis membrane with sufficient recovery (about 60%). Dialysates were collected by a sampling loop and introduced by an on-line injector to a microbore liquid chromatographic system for analysis of catecholamines. This assay yielded a detection limit of 0.2-0.5 pg/injection with acceptable intra- and inter-assay reproducibilities in 5 microl of dialysates. To evaluate the on-line microdialysis system, PC-12 cells were cultured in a petri dish within an incubator. The baseline concentration of dopamine in PC-12 cell culture medium was about 0.29 ng/ml which was elevated to 2.43 ng/ml after treatment with 0.5 mM potassium cyanide. In conclusion, the present microassay provides for the sensitive, direct measurement of catecholamines in culture medium while minimizing pretreatment procedures for sample preparation.

Animals↗
PubMed1976

Effect of diet on urinary excretion of VMA, HVA, metanephrine, and total free catecholamine in normal preschool children.

Quantitation of the urinary metabolites of catecholamines, including VMA, HVA, and metanephrines, from six normal preschool children was performed during a normal diet, a restricted diet, and a diet with increased amounts of vanilla, vanillin, and phenolic acids. Ingestion of these substances has been suspected of producing elevated values of urinary catecholamines and their metabolites. Urine was collected on the fourth day of each diet in two consecutive 12-hour aliquots, beginning at 8:00 AM. Statistically significant diurnal variation in excretion of all three metabolites and total free catecholamines (epinephrine, norepinephrine, and dopamine) was demonstrated. Diet did not alter exceretion of total free catecholamines or any of the three metabolites. This study suggests that a three-day restricted diet is not necessary prior to screening children for neuroblastoma when using quantitative assay methods and that all screening tests should be performed on a 24-hour urine sample.

Catecholamines↗
PubMed1988

Catecholamine metabolism in pheochromocytoma and normal adrenal medullae.

The tissue contents of catecholamine, its precursor and its major metabolites were determined in 8 human pheochromocytomas and 26 normal adrenal glands. Pheochromocytomas contained significantly larger amounts of norepinephrine, dopamine, dihydroxyphenylalanine, tyrosine hydroxylase activity, metanephrine, normetanephrine and vanillylmandelic acid than did normal adrenal medullae. The content ratio of epinephrine/norepinephrine in normal adrenal medullae was significantly higher than that in pheochromocytomas but there were considerable individual variations in the metanephrine/normetanephrine and vanillylmandelic acid/3-methoxy-4-hydroxyphenylethylglycol ratios in pheochromocytomas. In normal adrenal medullae the tissue content of tyrosine hydroxylase activity correlated inversely with the tissue contents of epinephrine (r equals -0.78, p less than 0.001), norepinephrine (r equals -0.78, p less than 0.001) and total catecholamines (r equals -0.87, p less than 0.001), respectively but no significant relation was found between both parameters in pheochromocytomas. These results indicate the possible presence of a negative feedback mechanism of catecholamine via tyrosine hydroxylase in normal adrenal medullae but none in pheochromocytomas. In addition, the increased degradation catecholamine pathway in pheochromocytomas appears to be unstable compared to that in normal adrenal medullae.

Adrenal Gland Neoplasms↗
PubMed2000

Ameliorating effects of amlodipine on plasma and myocardial catecholamines in BIO 53.58 Syrian hamsters, a model of dilated cardiomyopathy.

Our previous study has shown that the concentrations of norepinephrine, epinephrine and dopamine in the plasma of BIO 53.58 hamsters (a model of dilated cardiomyopathy: DCM) at 18 weeks of age (severe cardiomyopathic stage) were twice those of age-matched F1B control and conversely the myocardial norepinephrine level was decreased. The present study was undertaken to examine the effect of amlodipine on catecholamine concentration, myocardial receptors and histopathological changes in BIO 53.58 hamsters. Oral administration of amlodipine (10 mg/kg/day) for 7 weeks in 11 week-old-BIO 53.58 hamsters brought about marked decreases in the concentrations of norepinephrine, epinephrine and dopamine in the plasma, compared with those in vehicle-treated BIO 53.58 hamsters. This was accompanied by a concomitant increase in the concentration of myocardial catecholamine concentration. In other words, the concentrations of catecholamines in plasma and myocardium of amlodipine administered BIO 53.58 hamsters approximated to the control level in age-matched F1B. In addition, amlodipine administration caused a significant reduction of calcium deposition with a tendency toward a decrease in the myocardial necrosis, and it had little effect on the affinity and number of specific binding for (+)-[3H]PN 200-110, (-)-[125I]iodocyanopindolol (CYP) and [3H]prazosin in the myocardium. In conclusion, the present study shows that administration of amlodipine in BIO 53.58 hamsters may exhibit ameliorating effect on plasma and myocardial catecholamines with a significant reduction of calcium deposition. These data may offer further support for the use of amlodipine in patients with DCM.

Amlodipine↗
PubMed1999

Catecholamine concentration in rat liver after high level transection of the spinal cord.

The high level transection of the spinal cord (C-7) provokes a sustained increase of rat liver catecholamines: biphasic increase in norepinephrine level 1 hour and 24 hour after the operation and 7-fold increase of dopamine content 4 hour after the chordotomy. In contrast to cervical transection, sham operation causes only an initial catecholamine increase, the maximum being at the first hour after the surgery. Our experimental data indicate a possible participation of cervical spinal pathways in regulation of liver catecholamine content. It is also shown that bilateral adrenalectomy augments liver norepinephrine concentration in spinal rats as compared to the non-adrenalectomized ones. The results presented here indicate that cervical chordotomy affects the functioning of the sympatho-adrenal system, thus provoking specific changes in liver catecholamine content. The potential effect of such changes on a liver metabolic system (tyrosine aminotransferase induction) is discussed.

Adrenalectomy↗
PubMed2004

[Effect of modified liberation doxazosin on blood pressure, insulin resistance, and catecholamine secretion in patients with true resistant hypertension].

BACKGROUND AND OBJECTIVE: There are not studies about the utility of modified liberation doxazosin (MLD) in the combined treatment of the true resistant hypertension (TRH). The aim of this study was to determinate the effect of MLD on the blood pressure (BP), insulin resistance, and the catecholamine secretion in these patients. PATIENTS AND METHOD: Twenty seven patients with TRH from our unit were studied above and after 12 weeks of treatment with MLD (8 mg/day). The following parameters, among others, were analysed: office BP, ambulatory BP (ABPM), rennin, aldosterone and insulin plasmatic levels, urinary catecholamines and the insulin sensitivity index. RESULTS: Two patients were excluded because of adverse sides. After the treatment period, 53.8% of patients experimented a decrease of office diastolic BP higher than 10 mmHg, and 44% of them presented a decrease of 24-hour diastolic BP higher than 5 mmHg (answer patients). Likewise, 38.5% and 24% of patients achieved an adequate office BP and ABPM control, respectively. Also, a significant decrease of insulin resistance and urinary catecholamines, and an increase of aldosterone in plasma were observed. The rest of parameters remained invariables. CONCLUSIONS: MLD could be useful in the TRH treatment, because this drug decreased significantly BP, insulin resistance and catecholamine secretion in these patients, with a good tolerance.

Aldosterone↗
PubMed1997

Sympathetic mediation of stress and aggressive competition: plasma catecholamines in free-living male tree lizards.

The sympathetic nervous system and adrenal catecholaminergic tissue act to prepare an animal for "fight or flight" by release of catecholamines into synapses and plasma. However, few studies have measured plasma catecholamines in nonmammalian vertebrates and none have measured them in free-living animals. We report plasma levels of norepinephrine (NE), epinephrine (EPI) and dopamine (DA) in free-living tree lizards (Urosaurus ornatus) bled in the field: 1) immediately after capture, 2) after 10 minutes of restraint, 3) immediately after a staged territorial encounter and 4) four minutes after the end of a staged territorial encounter. Time to capture and time to bleed after capture were also recorded for each sample. Time to capture had little effect on plasma catecholamines suggesting that plasma catecholamines do not rise during the brief pursuit by the investigator necessary to capture the lizards in the field. In contrast, plasma NE and EPI increased during blood collection. However, this response was very consistent allowing comparisons to be made using analysis of covariance to control for time to bleed. Results indicate that restraint stress caused a large increase in plasma levels of NE, E and DA. Plasma levels of NE and E, but not plasma DA, were also significantly elevated both immediately and 4 minutes after aggressive encounters, although less so than following restraint stress. These studies show that stress and aggression activate the sympatho-adrenal system. This activation could be involved in behavioral changes during aggression, meeting metabolic demands of the encounter, or both.

Adrenal Glands↗
PubMed2001

Identification and quantification of catecholamines in potato plants (Solanum tuberosum) by GC-MS.

Dopamine, norepinephrine, and normetanephrine were identified by GC-MS in potato (Solanum tuberosum L.) plants, the latter was new for plants. The highest amount of catecholamines was found in leaves. A developmental stage dependent variation in potato leaf catecholamines accumulation was also observed with highest level in third leaves. Catecholamine contents decrease during cold storage of tubers to undetectable levels. Mechanical wounding of leaves led to a small increase in the level of catecholamines investigated.

Catecholamines↗