Norepinephrine and neuropeptide Y increase intracellular Ca2+ in cultured porcine aortic smooth muscle cells.

We used the fluorescent Ca2+ indicator Fura-2 in cultured porcine aortic smooth muscle cells (PASMC) to study effects of the sympathetic neurotransmitters norepinephrine (NE) and neuropeptide Y (NPY) on free intracellular Ca2+ (Cai). Both transmitters transiently increased intracellular Ca2+ in a concentration-dependent manner. Selective agonists and antagonists demonstrated that the NE-stimulated Cai increase is predominantly (if not exclusively) mediated by alpha 2-adrenoceptors, whereas the NPY response appears to be mediated by the peptide YY-insensitive Y3-like receptor subtype. Pretreatment of cells with pertussis toxin abolished NPY and alpha-adrenoceptor agonist-stimulated intracellular Ca2+ elevations (but not those stimulated by angiotensin II) suggesting involvement of a Gi-like G-protein. alpha 2-Adrenoceptor-stimulated Ca2+ increases resulted from mobilization from intracellular stores, whereas Y3-like NPY receptors mobilized Ca2+ from intracellular stores and also promoted Ca2+ influx.

[N2O-supplemented intravenous anesthesia versus inhalation anesthesia. A comparative study of the sympathoadrenergic reaction and postoperative vigilance]. / N2O-supplementierte, intravenöse Anästhesie versus Inhalationsanästhesie. Vergleichende Untersuchung der sympathoadrenergen Reaktion und der postoperativen Vigilanz.

The aim of the present study was a comparison of the intraoperative sympathoadrenergic response and the postoperative vigilance of a propofol/alfentanil anaesthesia to a conventional isoflurane anaesthesia. 25 patients were admitted to the study undergoing septorhino surgery. Patients with continuous intravenous anaesthesia with propofol/alfentanil combined with nitrous oxide showed better haemodynamic conditions without an increase of blood pressure and catecholamines under laryngoscopy, intubation and surgical stimulation. In contrast to that the patients with isoflurane anesthesia showed a significant increase in haemodynamic parameters and capillary bloodflow. The measured plasma adrenalin levels showed wide intraindividual fluctuation but no significant difference between the groups. The suppression of plasma noradrenaline was more pronounced under intravenous anaesthesia. Recovery was significantly faster and vigilance significantly better in the patients undergoing intravenous anaesthesia. After 30 min patients with i.v. anaesthesia fulfilled all the conditions to be transferred to the regular ward; the other group needed more than one hour. It can be concluded that continuous i.v. anaesthesia with propofol/alfentanil is superior in suppressing the stress response to invasive stimuli and provides faster recovery and better postoperative analgesia.

Development of hypertension in neuroblastoma during therapy: a case report.

A case of stage 4 neuroblastoma that developed excessive hypertension on day 120 of chemotherapy is presented. The tumor initially had responded well to chemotherapy; however, while the tumor mass decreased, plasma and urine catecholamines and the blood pressure increased. The plasma concentrations of noradrenaline, adrenaline, and dopamine increased to 26.4, 1.8, and 36.2 micrograms/l, respectively. The profile of catecholamine metabolites changed: on day 150 of therapy, noradrenaline, adrenaline, and dopamine levels were increased, whereas HVA and VMA levels were decreased when compared to day 1 of therapy. The only residual neuroblastoma tissue visible on MIBG scintigraphy on day 150 of treatment was a metastasis in the left tibia which was irradiated with 24 Gy. The adrenaline concentration in the left femoral vein was twice as high compared to the right femoral vein. A treatment, possibly radiation-associated tumor cell alteration resulting in a different catecholamine production, is discussed.

Terbutaline-induced desensitization of beta 2-adrenoceptor in vivo function in humans: attenuation by ketotifen.

Use of beta-adrenoceptor agonists in long-term treatment of patients with chronic asthma bronchiale or heart failure is of limited value because beta-adrenoceptor desensitization develops. The antiallergic drug ketotifen prevents beta-adrenoceptor agonist-induced desensitization of rat and human pulmonary and lymphocyte beta 2-adrenoceptors. In 10 healthy volunteers in a double-blind, placebo-controlled study, we investigated whether ketotifen also prevents beta-adrenoceptor agonist-induced desensitization of beta 1- and/or beta 2-adrenoceptor-mediated physiologic in vivo effects. beta 1-Adrenoceptor-mediated effects were isoprenaline (ISO) infusion-induced increase in systolic blood pressure (SBP) and bicycle exercise-induced increase in heart rate (HR); beta 2-adrenoceptor-mediated effects were ISO infusion-induced increase in plasma norepinephrine (NE) and decrease in diastolic blood pressure (DBP); ISO infusion-induced increase in HR was assessed as mixed beta 1- and beta 2-adrenoceptor-mediated effect. These parameters were assessed before and after a 14-day treatment with the beta 2-adrenoceptor agonist terbutaline (5 mg three times daily) with or without simultaneous administration of ketotifen (1 mg twice daily). Terbutaline desensitized all in vivo effects involving beta 2-adrenoceptors (ISO-induced decrease in DBP and increase in plasma NE and, to a minor extent, the mixed beta 1- and beta 2-adrenoceptor-mediated increase in HR), but did not affect beta 1-adrenoceptor-mediated in vivo effects; concomitant treatment of the volunteers with ketotifen markedly blunted terbutaline-induced desensitization of beta 2-adrenoceptor in vivo function. We conclude that ketotifen prevents, or at least attenuates, beta-adrenoceptor agonist-induced desensitization of beta 2-adrenoceptor in vivo function.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of insulin-induced hypoglycemia on beta 2-adrenoceptor density and proliferative responses of human lymphocytes.

We studied the effects of insulin (0.1 IU/kg BW, iv)-induced hypoglycemia on lymphocyte beta 2-adrenoceptor function, lymphocyte subset distribution, and proliferative response to mitogen stimulation in 10 healthy volunteers. Thirty minutes after insulin injection plasma glucose levels were markedly decreased; concomitantly, plasma epinephrine levels had increased about 10-fold; plasma norepinephrine levels, however, increased only moderately. Lymphocyte beta 2-adrenoceptor density and the cAMP response to 10 mumol/L isoproterenol stimulation were elevated; lymphocyte Ts/c-cells had increased, whereas Th-cells had decreased, resulting in a decrease in the Th-/Ts/c-cell ratio from 1.7 to 1.0. These changes were accompanied by a significantly reduced lymphocyte proliferative response (measured as [3H]thymidine uptake) to mitogen stimulation. Two hours after insulin injection plasma catecholamines, lymphocyte beta 2-adrenoceptor function, lymphocyte subset distribution, and proliferative responses had returned to nearly preinsulin levels. We conclude that acute vigorous increases in endogenous epinephrine evoked by insulin-induced hypoglycemia are associated with increases in lymphocyte beta 2-adrenoceptor function, redistribution of lymphocyte subsets, and an (at least transiently) attenuated in vitro immune responsiveness.

Adrenaline and noradrenaline as possible chemical mediators in the pathogenesis of arteriosclerosis.

We studied the relationships between adrenaline and noradrenaline and factors associated with arteriosclerosis to determine whether catecholamines contribute to the atherogenetic process. We investigated the effects of adrenaline and noradrenaline on cultures of vessel wall cells from rats and analyzed plasma catecholamine levels in humans exposed to atherogenic risk factors, undergoing hemodialysis treatment or following myocardial infarction or stroke. I. Cultured endothelial and smooth muscle cells from vessel walls exhibited enhanced proliferation when exposed to adrenaline or noradrenaline. This indicates that catecholamines trigger the activation of vascular wall cells in vitro. Such activation, the unspecific mesenchymal reaction, is the predominant characteristic change in early atherogenesis. II. In individuals subjected to the atherogenic risk factors smoking, essential hypertension and mental stress, plasma adrenaline concentrations were statistically significantly elevated. Mental stress also caused significantly elevated plasma noradrenaline levels. Plasma noradrenaline concentrations were also elevated in smoking and hypertensive individuals when compared with certain controls, but the differences failed to be statistically significant. III. In dialysis patients, plasma adrenaline and noradrenaline concentrations showed a positive correlation with the activity of the sclerotic process; i.e., plasma catecholamine concentrations increased with the severity of the disease. IV. Patients with persisting arteriosclerotic vascular disease, i.e., patients who had had a myocardial infarction or stroke, had significantly elevated plasma adrenaline and/or noradrenaline levels as late as one year after the event. The results of our investigations suggest that adrenaline and noradrenaline may act as chemical mediators during atherogenesis in man, thus contributing to the development and subsequent complications of arteriosclerosis.

[Functional differentiation of the vascular endothelium in high risk pregnancies]. / Funktionelle Differenzierung des vaskulären Endothels bei Hochrisikoschwangerschaften.

Endothelial cell functions regulating fetal hemodynamics and placental perfusion are modulated by metabolites of arachidonic acid, in particular derivatives of cyclooxygenase such as prostaglandins. We utilized an in vitro system to study the possible role of the vascular endothelium in the pathogenesis of chronic placental insufficiency. Cellular growth and prostaglandin metabolism were investigated in cultured umbilical vein endothelial cells from mothers exposed to risk factors such as smoking and diabetes mellitus during pregnancy. The study comprised cells from smoking (n = 18) and diabetic mothers (n = 5) compared to non-smoking, non-diabetic controls (n = 20). Endothelial cells from smoking and diabetic mothers grew less readily than did control cells. The synthesis of the prostaglandins PGI2 and PGE2, both potent vasodilators and platelet aggregation inhibitors, was significantly reduced in endothelial cells from smoking mothers and from mothers suffering from diabetes of various stages. Thus, reduced prostaglandin synthesis may be a cause of impaired placental perfusion in high risk pregnancies. Our data suggest that prostaglandins may play an important role in the etiology and pathogenesis of chronic placental insufficiency.

A possible role of catecholamines in atherogenesis and subsequent complications of atherosclerosis.

Cultured smooth muscle cells (SMC) from rat aorta and endothelial cells (EC) from pig aorta were used to study the effect of the catecholamines epinephrine and norepinephrine on cell proliferation. Both stimulated growth of SMC and EC when added to the culture medium. Besides epinephrine and norepinephrine, dopamine and some of their metabolites also stimulated proliferation of cultured endothelial cells. Smooth muscle cells originating from rats being exposed to atherosclerotic risk factors, like diabetes, hypertension and balloon-injury, exhibited an increased susceptibility to these catecholamines compared to SMC from control animals. In comparison to normotensive control animals a 10-fold elevated plasma concentration of epinephrine was found in hypertensive rats. In man plasma epinephrine and norepinephrine concentration was determined in a healthy control group and in patients suffering from diabetes mellitus and coronary artery disease. Plasma epinephrine and norepinephrine levels were similar in patients suffering from diabetes mellitus compared to the control group. But in patients with coronary artery disease significantly higher plasma concentrations for epinephrine (p less than 0.001) and norepinephrine (p less than 0.01) were observed. These data support the hypothesis that catecholamines may play a role in the development and subsequent complications of atherosclerosis.

[A single, rapid, selective and quantitative determination of adrenaline and noradrenaline in the plasma by a combination of solvent extraction, HPLC separation and electrochemical detection]. / Einfache, schnelle, selektive und quantitative Bestimmung von Adrenalin und Noradrenalin im Plasma durch Kombination von Flüssigextraktion, HPLC-Trennung und elektrochemischer Detektion.

A very simple and rapid solvent extraction system for the selective and quantitative isolation of epinephrine and norepinephrine from plasma is described. The extraction system makes use of the complex formation in alkaline medium between diphenylborate and the diol group of the catecholamines in combination with ion-pair formation. The extraction procedure, in conjunction with HPLC-separation and electrochemical detection, allows the quantitative determination of epinephrine and norepinephrine from plasma. This method is a specific as the commonly used adsorption on alumina, but has a much better recovery and even greater precision, especially for the quantitative determination of epinephrine.

A double antibody sandwich microELISA for measuring human platelet factor 4.

With the aim of investigating patients with a higher risk of thrombosis we developed a method for determining platelet factor 4 (PF4) in human plasma. Using the double antibody sandwich ELISA technique we set up a test system that allows the determination of PF4 concentrations in plasma samples from 1 to 100 ng/ml. This method is more sensitive and as specific as commercially available RIA kits, but has the advantage of being cheaper and less time consuming. Furthermore the ELISA does not require radioactive materials. The complete reaction is carried out in microtiter wells, and an ELISA-reader connected to an Apple computer does all the calculations needed for quantitative measurements.