Microcirculation and haemodynamics after infraclavicular brachial plexus block using adrenaline as an adjuvant to lidocaine: a randomised, double-blind, crossover study in healthy volunteers.

We evaluated the effect of adrenaline on human skin microcirculation (nutritive and sub-papillary) and systemic cardiovascular variables after it was added to lidocaine in infraclavicular brachial plexus blocks. Twelve healthy, non-smoking male volunteers were included, each attending two study sessions 2 weeks apart, and they were studied using a crossover design. In both sessions, they received an ultrasound-guided infraclavicular brachial plexus block in the non-dominant arm with 0.4 ml.kg-1 lidocaine, 15 mg.ml-1 with or without adrenaline 5 µg.ml-1 . Microcirculation was assessed by laser Doppler fluxmetry (sub-papillary blood flow), capillary video microscopy (nutritive blood flow) and continuous temperature measurements. Heart rate and arterial pressure were recorded continuously and non-invasively. Median (IQR [range]) sub-papillary blood flow increased substantially 30 min after the brachial plexus block, from 8.5 (4.4-13.5 [2.9-28.2]) to 162.7 (111.0-197.8 [9.5-206.7]) arbitrary units with adrenaline (p = 0.017), and from 6.9 (5.3-28.5 [1.8-42.1] to 133.7 (16.5-216.7 [1.0-445.0] arbitrary units without adrenaline (p = 0.036). Nutritive blood flow (functional capillary density, capillaries.mm-2 , measured at the dorsal side of the hand) decreased in the blocked extremity when adrenaline was used as adjuvant, from median (IQR [range]) 45 (36-52 [26-59]) to 38 (29-41 [26-42]), p = 0.028, whereas no significant change occurred without adrenaline. Median finger skin temperature (°C) increased by 44% (data pooled) with no significant differences between the groups. No significant changes were found in the systemic cardiovascular variables with or without adrenaline. We conclude that lidocaine infraclavicular brachial plexus blocks caused an increase in skin sub-papillary blood flow. The addition of adrenaline produced stronger and longer lasting blocks, but decreased the nutritive blood flow.

Dysfunctional arteriovenous anastomoses in hands of systemic sclerosis patients with digital ulcers.

OBJECTIVES: Previous studies indicate that the arteriovenous anastomoses (AVAs) and the arterioles with the nutritive flow are involved in the pathophysiologic process disturbing hand blood flow in systemic sclerosis (SSc). However, impact of different part of the microvascular system involved in digital ulcers (DU) is not well known. Here, we aimed to assess the vasomotor activity of the AVAs in the hands of patients with and without DU in SSc. METHODS: Simultaneous recordings were made of laser Doppler flux in the finger pulp and thenar eminence, together with ipsilateral radial artery blood velocity and mean arterial blood pressure (MAP) in 22 non-smoking SSc patients and 13 aged-matched healthy controls. RESULTS: AVA responses in the finger pulp to spontaneous vasoconstrictor nerve impulses were abolished in 64% of the SSc patients. Correlation and cross-spectra analysis showed positive correlation between blood flow changes and MAP changes, indicating a passive vascular bed in the SSc finger pulp with blood flow variations depending on short-term variability in MAP. Dysfunctional AVAs were identified in all the patients with a history of DU (n=8), while none of the patients with normal AVA function had episodes of DU (n=8) (p= 0.017). CONCLUSIONS: We found that in SSc patients with DU there is a dysfunction of the AVAs of the finger pulp. This proof-of-concept study supports the notion that AVA dysfunction may play a critical role in SSc related DU. AVA dysfunction may be a part of autonomic dysfunction in SSc.

Changes in pulmonary function during pregnancy: a longitudinal cohort study.

OBJECTIVE: To record any physiological changes in lung function during healthy pregnancies, and evaluate the influence of parity, pregestational overweight, and excessive weight gain. DESIGN: Longitudinal cohort study. SETTING: Antenatal clinic at Oslo University Hospital. POPULATION: One hundred healthy white women with singleton pregnancies. METHODS: The women were studied with repeated measures of lung function using spirometry at a gestational age of 14-16, 22-24, 30-32, and 36 weeks, and at 6 months postpartum. MAIN OUTCOME MEASURES: Forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1), and peak expiratory flow (PEF), also expressed as a percentage of predicted values according to age and height: i.e. FVC%, FEV1%, and PEF%. RESULTS: Both FVC and FVC% increased significantly after 14-16 weeks of gestation (P=0.001), as was the case for both PEF and PEF% (P<0.001). FVC, FVC%, PEF, and PEF% in early and mid-pregnancy were significantly lower compared with the postpartum value (all P<0.05). Nulliparous women had an overall 4.4% lower value of FVC% than parous women (P=0.039). There were no differences in FVC, FEV1, or PEF dependent upon pregestational overweight or excessive weight gain. CONCLUSIONS: Forced vital capacity (FVC) increases significantly after 14-16 weeks of gestation. The FVC% is significantly higher in parous compared with primigravida women, suggesting that the changes in FVC occurring during pregnancy persist postpartum. PEF increases significantly during healthy pregnancies, and should be interpreted cautiously in pregnant women with impaired lung function.

Does cardiovascular autonomic dysfunction contribute to fatigue in myasthenia gravis?

Myasthenia gravis (MG) is an autoimmune disease characterized by fatigable muscle weakness. Despite full spontaneous or pharmacological remission some MG patients still complain of physical and mental fatigue. Fatigue has been related to autonomic dysregulation. The aim of this study was to assess autonomic responses in a group of MG patients in complete remission but complaining of persistent fatigue. Seventeen well-regulated but persistently fatigued MG patients and 17 individually matched controls underwent echocardiography assessing systolic and diastolic heart function. Beat to beat cardiovascular responses at rest and to 30o head-up tilt, tilt-back, and 2-min static handgrip contraction were recorded. Fatigued MG patients had a statistically significant higher resting HR than their matched controls (p=0.03). The difference in resting heart rate between MG patients not using acetylcholine esterase inhibitors (AChEi) and their matched controls was even more pronounced (p=0.007). The autonomic cardiovascular adjustments to head-up tilt, tilt-back and handgrip contraction were not statistically significant different between patients and controls. We found a higher resting heart rate in all well-regulated but fatigued MG patients compared with controls. The difference was more pronounced between patients not taking AChEi compared to their matched controls. This finding may reflect a disturbed resting sympathovagal balance and this might be a contributing factor to the fatigue symptoms.

Does the great saphenous vein stripping improve arterial leg blood flow during exercise?

OBJECTIVES: It has been shown that the leg muscle pump increases arterial leg blood flow during upright exercise in healthy subjects, and that this effect is reduced in patients with incompetence of the great saphenous vein (GSV). In this study, patients with GSV reflux causing varicose veins were investigated after GSV stripping, to see whether the muscle pump effect on arterial leg blood flow is improved. DESIGN: Prospective case study. METHODS: Nine patients with GSV incompetence resulting in symptomatic varicose veins, but without peripheral artery disease were included in this study. Patients exercised in the supine and 30° head up tilted positions by rhythmically pressing down a pedal with one foot. Blood flow was measured in the femoral artery using Doppler ultrasound. The Exercise-induced rise in femoral artery blood flow was compared in the supine and 30° head up tilted positions. Patients were investigated both before and after undergoing saphenofemoral ligation and GSV stripping as a treatment for their varicose veins. The arterial blood flow response to exercise was compared between the pre and postoperative observations. RESULTS: Prior to GSV stripping the immediate rise in femoral flow was 0.25 l min(-1) above rest in both supine and tilted positions. After GSV stripping however, the rise in flow was 30% larger in the tilted position than in the supine position (0.26 vs. 0.20 l min(-1), P < 0.05). CONCLUSIONS: GSV stripping modestly improves arterial leg blood flow at the onset of exercise in patients with GSV insufficiency, because of an improved effect of the leg muscle pump.

In vivo measurement of haloperidol affinity to dopamine D2/D3 receptors by [123I]IBZM and single photon emission computed tomography.

This study examines the feasibility of a steady-state bolus-integration method with the dopamine D2/D3 receptor single photon emission computer tomography (SPECT) tracer, [123I]IBZM, for determination of in vivo affinity of haloperidol. The nonspecific binding of [123I]IBZM was examined in the rat brain by infusion of haloperidol to plasma levels approximately 100 times the Kd level in man. In humans, Kd for haloperidol binding was measured in four healthy volunteers that were examined twice: once with partial dopamine D2/D3 receptor blockade obtained by a scheduled infusion of unlabeled haloperidol (0.7 mg total dosage), and once in an unblocked state. Blood sampling and SPECT were performed intermittently during 6 hours after intravenous [123I]IBZM bolus injection. Plasma [123I]IBZM was determined by octane extraction. Plasma haloperidol was determined by a radioimmunoassay, and plasma protein binding was determined by equilibrium dialysis. In humans, the striatal D2/D3 receptor occupancy was 0.27+/-0.085 and the in vivo Kd for haloperidol was 0.25+/-0.1 nmol/L, which is comparable to Kd values as obtained from in vitro studies. The authors conclude that steady-state [123I]IBZM SPECT studies allow for determination of dopamine D2/D3 receptor occupancy in striatum and in vivo measurement of drug affinity to striatal dopamine D2 and D3 receptors.

Tyrosine hydroxylase and Parkinson's disease.

A consistent neurochemical abnormality in Parkinson's disease (PD) is degeneration of dopaminergic neurons in substantia nigra, leading to a reduction of striatal dopamine (DA) levels. As tyrosine hydroxylase (TH) catalyses the formation of L-DOPA, the rate-limiting step in the biosynthesis of DA, the disease can be considered as a TH-deficiency syndrome of the striatum. Similarly, some patients with hereditary L-DOPA-responsive dystonia, a neurological disorder with clinical similarities to PD, have mutations in the TH gene and decreased TH activity and/or stability. Thus, a logical and efficient treatment strategy for PD is based on correcting or bypassing the enzyme deficiency by treatment with L-DOPA, DA agonists, inhibitors of DA metabolism, or brain grafts with cells expressing TH. A direct pathogenetic role of TH has also been suggested, as the enzyme is a source of reactive oxygen species (ROS) in vitro and a target for radical-mediated oxidative injury. Recently, it has been demonstrated that L-DOPA is effectively oxidized by mammalian TH in vitro, possibly contributing to the cytotoxic effects of DOPA. This enzyme may therefore be involved in the pathogenesis of PD at several different levels, in addition to being a promising candidate for developing new treatments of this disease.

Peripheral vasoconstriction shortly after onset of moderate exercise in humans.

The immediate cardiovascular responses at the onset of supine dynamic leg exercise were studied by noninvasive methods in healthy humans. Total peripheral conductance (TPC), heart rate, and cardiac output increased very rapidly at the onset of exercise. Mean arterial pressure (MAP) showed a moderate anticipatory increase during a 10-s countdown to exercise and then decreased (but not below resting level) during the first 10 s of exercise. The TPC response was biphasic, and TPC started to fall from its peak value approximately 12 s after onset of exercise. This peripheral vasoconstriction increased MAP. After 25 s, the cardiovascular variables were stable for the rest of the 2-min exercise period. In the same subjects, cholinergic blockade was induced by atropine sulfate (0.035 mg/kg) and resting cardiac output, MAP, and TPC increased considerably. The exercise protocol was repeated after atropine, and the increase in heart rate at onset of exercise was slower and smaller. MAP decreased and remained depressed throughout the exercise period. A monophasic increase in TPC was seen. We suggest that, in the normal situation, the biphasic response in TPC reflects a baroreflex sympathetic vasoconstriction very shortly after onset of exercise and that this response is due to a rapid increase in set point for arterial pressure control at the onset of exercise. After cholinergic blockade, MAP was probably continuously well above the set point for arterial pressure control both before and during exercise and no reflex vasoconstriction was observed in this situation.

Midodrine prevents orthostatic intolerance associated with simulated spaceflight.

Many astronauts after being weightless in space become hypotensive and presyncopal when they assume an upright position. This phenomenon, known as orthostatic intolerance, may interfere with astronaut function during reentry and after spaceflight and may limit the ability of an astronaut to exit a landed spacecraft unaided during an emergency. Orthostatic intolerance is more pronounced after long-term spaceflight and is a major concern with respect to the extended flights expected aboard the International Space Station and for interplanetary exploration class missions, such as a human mission to Mars. Fully effective countermeasures to this problem have not yet been developed. To test the hypothesis that alpha-adrenergic stimulation might provide an effective countermeasure, we conducted a 16-day head-down-tilt bed-rest study (an analog of weightlessness) using normal human volunteers and administered the alpha(1)-agonist drug midodrine at the end of the bed-rest period. Midodrine was found to significantly ameliorate excessive decreases in blood pressure and presyncope during a provocative tilt test. We conclude that midodrine may be an effective countermeasure for the prevention of orthostatic intolerance following spaceflight.

Lactate release, concentration in blood, and apparent distribution volume after intense bicycling.

To study the release of lactate from muscle and its relationship to the blood lactate concentration during and after intense bicycling, young men cycled at 5.5 W kg(-1) body mass for 2 min to exhaustion or stopped after 1 min (nonexhaustive ride). The leg's release of lactate during and after each ride was taken from the measured blood flow and lactate concentrations in arterial and femoral-venous blood. Muscle biopsies were taken in separate experiments and analyzed for lactate. During the bicycling, 6 to 10% of the lactate produced was released to the blood. During exercise and for the first few minutes after, the rate of lactate release did not differ between 2 min exhaustive and 1 min nonexhaustive bicycling. The integrated release (exercise plus recovery) for the 1 min bicycling was 60 to 80% of the corresponding value of the 2 min exhaustive bicycling. In the late recovery, the blood lactate concentration was 3 to 5 times higher after 2 min exhaustive bicycling than after the 1 min nonexhaustive bicycling. There was thus a mismatch between the amount of lactate released and measured concentration in blood, reflecting a smaller distribution volume after the exhaustive bicycling. The blood lactate concentration may therefore not be a good measure of the lactate production and anaerobic energy release during bicycling.

Recording of short-term finger blood pressure changes induced by an arterial occlusive thigh cuff: comparison between the modified oscillometric and Finapres techniques.

Blood pressure recordings obtained by the volume clamp method (Finapres monitor, Ohmeda, USA) were compared with measurements by using a novel modification of the oscillometric method (UT9201 beat-to-beat monitor, University of Tartu, Estonia). Step changes in total peripheral resistance were introduced in 11 healthy volunteers by rapid inflation to the suprasystolic pressure of bilateral thigh cuffs during a rhythmical exercise with the quadriceps muscles and simultaneously performed handgrip compression. The provoked short-term changes in mean arterial pressure (MAP) were similarly tracked by both devices. The agreement between the methods was better for MAP rise than drop. The group-averaged difference UT9201 minus Finapres did not exceed 1.2 mmHg (NS, p = 0.46) after cuff successive inflation but yielded 3.8 mmHg (significant, p < 0.05) after cuff successive deflation.

Does venous insufficiency impair the exercise-induced rise in arterial leg blood flow?

OBJECTIVES: It has been shown that the leg muscle pump increases the immediate rise in arterial leg blood flow during upright exercise in healthy subjects. The present study is the first to investigate the muscle pump effect in exercise hyperaemia in patients with venous insufficiency, who should be lacking an optimally functioning muscle pump. METHODS: Any muscle pump effect is more pronounced in an upright position because of gravitation. The exercise-induced rise in femoral artery flow (FF) (ultrasound Doppler) was thus compared in the supine and 30° head-up tilted position in 10 patients. RESULTS: Neither the transient nor the steady-state rise in FF showed any difference between positions. This is in contrast to the previous findings in healthy subjects, where the transient rise in FF was larger in the tilted position. CONCLUSION: The muscle pump effect in exercise hyperaemia seems to be reduced or lacking in these patients.

Handgrip contraction induces a linear increase in arterial pressure by peripheral vasoconstriction, increased heart rate and a decrease in stroke volume.

AIM: The hypothesis that isometric handgrip induces a progressive increase in arterial pressure and a linear increase in setpoint for arterial pressure control was tested. METHODS: The continuous time course of changes in heart rate (HR), stroke volume (SV) and mean arterial pressure (MAP) was recorded during a 2-min handgrip contraction of 40% of maximal voluntary contraction force. Twice during the development of the handgrip-induced, gradual pressure increase of ∼25 mmHg, additional, transient changes in arterial pressure were mechanically induced. The subsequent baroreflex responses to these additional pressure changes were studied. The additional steep increase in arterial pressure (∼10 mmHg) was induced both after 70 and 100 s of handgrip contraction, by inflating bilateral thigh cuffs to suprasystolic pressure. Cuff pressure was released after 10s, thus introducing a steep decrease in MAP. RESULTS: During the development of the handgrip-induced pressure increase, HR increased, SV decreased, cardiac output (CO) increased slightly and total peripheral conductance (TPC=CO/MAP) increased (i.e. peripheral vasoconstriction). The circulatory responses to the additional, sudden increase and subsequent decrease in arterial pressure after 70 and 100 s perfectly adjusted arterial pressure back to the linear increase in MAP, indicating an effective baroreflex response. CONCLUSION: The increase in MAP which characterizes handgrip-induced pressure response can be regarded as a result of a gradual increase in the set point of the arterial baroreflexes, with no change in the time course and magnitude of the baroreflex responses to additional, induced changes in MAP.

Stroke volume decreases during mild dynamic and static exercise in supine humans.

AIM: The contributions of cardiac output (CO) and total peripheral resistance to changes in arterial blood pressure are debated and differ between dynamic and static exercise. We studied the role stroke volume (SV) has in mild supine exercise. METHODS: We investigated 10 healthy, supine volunteers by continuous measurement of heart rate (HR), mean arterial blood pressure, SV (ultrasound Doppler) and femoral beat volume (ultrasound Doppler) during both dynamic mild leg exercise and static forearm exercise. This made it possible to study CO, femoral flow (FF) and both total and femoral peripheral resistance beat-by-beat. RESULTS: During a countdown period immediately prior to exercise, HR and mean arterial pressure increased, while SV decreased. During mild supine exercise, SV decreased by 5-8%, and most of this was explained by increased mean arterial pressure. Dynamic leg exercise doubled femoral beat volume, while static hand grip decreased femoral beat volume by 18%. FF is tightly regulated according to metabolic demand during both dynamic leg exercise and static forearm exercise. CONCLUSION: Our three major findings are, firstly, that SV decreases during both dynamic and static mild supine exercise due to an increase in mean arterial pressure. Secondly, femoral beat volume decreases during static hand grip, but FF is unchanged due to the increase in HR. Finally, anticipatory responses to exercise are apparent prior to both dynamic and static exercise. SV changes contribute to CO changes and should be included in studies of central haemodynamics during exercise.

Lactate elimination and glycogen resynthesis after intense bicycling.

OBJECTIVE: Muscles break down glycogen to lactate during intense exercise, and in the recovery period, glycogen reappears while lactate disappears. The purpose of this study was to examine to what extent lactate is resynthesized to glycogen within the formerly active muscles themselves in man. MATERIAL AND METHODS: Fifteen healthy young men cycled for 2 min to exhaustion. Muscle biopsies were taken from the knee extensor muscle before the exercise, just after the ride, and again after 45 min of recovery. In addition, blood samples were taken from the femoral artery and vein, and the leg blood flow was measured using the ultrasound Doppler technique. The muscle biopsies were analysed for glycogen, lactate and other metabolites, and the blood samples were analysed for lactate and glucose. The exchanges of lactate and glucose of the leg were assessed by multiplying the measured arterio-venous (a-v) differences by the blood flow. RESULTS: During the exercise the muscles broke down 20+/-4 mmol glycogen kg(-1) wet muscle mass and produced 26+/-1 mmol lactate kg(-1). In the recovery period after 24+/-1 mmol lactate kg(-1) had disappeared, of which 48 % was released to the blood, 52 % disappeared within the muscle. An R-value of 0.62 across the leg suggests that none of the lactate was oxidized. Altogether, 10+/-3 mmol glycogen kg(-1) reappeared during recovery. Glucose uptake accounted for 2 mmol kg(-1) and glycolytic intermediates (G-6-P and free glucose) accounted for 4 mmol kg(-1); 4 mmol glycogen kg(-1) (42 %) reappeared from unknown sources. CONCLUSIONS: The present data are compatible with the idea that around half of the lactate produced during intense bicycling is resynthesized to glycogen within the working muscles themselves in the recovery period after the bicycling.

Respiration-synchronous fluctuations in stroke volume, heart rate and arterial pressure in humans.

1. Simultaneous recordings of beat-to-beat left cardiac stroke volume (SV, pulsed ultrasound Doppler), mean arterial pressure (MAP) and heart rate (HR) were obtained in ten healthy young adults during spontaneous respiration at supine rest, before and after cholinergic blockade by atropine (0.035 mg kg-1). 2. Respiration-synchronous fluctuations in SV, HR, cardiac output (CO) and MAP were quantified by spectral analysis of the recordings of each of these variables. 3. Before atropine administration, respiration-synchronous fluctuations in HR and SV were prominent. The changes in HR and SV were inversely related and variation in SV was the main source of respiratory variability in CO. Respiration-synchronous fluctuations in MAP were mainly caused by variations in CO. 4. After cholinergic blockade, respiratory HR variations were eliminated, whereas the respiratory fluctuations in SV persisted. The fluctuations in CO and MAP increased. In this situation, mechanically induced variations in SV were not counteracted by inverse HR fluctuations and the influence on CO thus increased. 5. The main source of respiratory fluctuations in MAP in supine humans is thus variation in SV, while inverse, vagally mediated HR variations tend to reduce the fluctuations in CO and MAP.

Short-term control of cardiovascular function: estimation of control parameters in healthy humans.

In a previous study, we recorded short-term cardiovascular responses after a steep increase in arterial pressure in healthy humans [Am. J. Physiol. 266 (Heart Circ. Physiol. 35): H199-H211, 1994]. The aim of the present study was to develop a mathematical model of the baroreflex control of arterial pressure, to use this model with the previously recorded data to estimate unknown parameters in the reflex control loop, and then to analyze the overall open- and closed-loop performance of the system by model simulations with use of individual sets of optimal parameters. The mathematical model consists of a heart, a linear elastic arterial reservoir, and two parallel resistive vascular beds. The arterial baroreflex loop is modeled by two separate time domain processing objects, each with its own gain, time constant, and delay, to simulate the action of a sympathetic signal to the peripheral vascular bed and a parasympathetic signal to the heart. In repeated model simulations, the control parameters in the model were systematically adjusted by an automated algorithm that minimized the deviations between the time courses of the cardiovascular variables simulated by the model and the previously recorded responses in each individual. In all 10 subjects, the short-term cardiovascular responses were adequately simulated by using individual sets of parameters in the model. Open-loop transfer functions for arterial pressure control were obtained by using the individual sets of optimal model parameters in new simulation runs. Open-loop gain for arterial pressure control at nearly zero frequency (steady state) was between 0.9 and 4. Model simulations also indicated an underdampened response at 0.05-0.07 Hz in the closed-loop situation in four subjects, corresponding to peaks in the mean arterial pressor power spectra obtained from separate recordings of spontaneous variations in the resting situation.

[Digestive system's large and changing needs of blood supply]. / Fordøyelseskanalens store og vekslende behov for blodtilførsel.

Doppler utrasonography has made it possible to record blood flow to the digestive tract (the superior mesenteric artery) directly and continuously in unanaesthetized, healthy humans. Several research groups have demonstrated how blood flow to the tract increases gradually and markedly after a meal, and more so after a big meal than after a small one. The increase in blood flow reaches its maximum after 20-40 minutes and lasts for 1.5-2 hours. In the postprandial period there is a parallel and similar increase in cardiac output; the meal thus imposes an increased work load on the heart. Carbohydrate meals, as well as meals of protein or fat all release increases in local blood flow as well as in cardiac output. Surprisingly, during physical exercise of relatively high intensity, there is no reduction in blood flow to the digestive tract in humans. This is in contrast to the exercise-caused flow reduction observed in several animal species. In the postprandial state the large increase in cardiac output caused by muscular exercise is actually added to the increase already established by the meal. This course of events helps to explain why patients with angina pectoris are more prone to chest pains after a meal.

Interaction of phosphorylated tyrosine hydroxylase with 14-3-3 proteins: evidence for a phosphoserine 40-dependent association.

Tyrosine hydroxylase (TH) has been reported to require binding of 14-3-3 proteins for optimal activation by phosphorylation. We examined the effects of phosphorylation at Ser19, Ser31 and Ser40 of bovine TH and human TH isoforms on their binding to the 14-3-3 proteins BMH1/BMH2, as well as 14-3-3 zeta and a mixture of sheep brain 14-3-3 proteins. Phosphorylation of Ser31 did not result in 14-3-3 binding, however, phosphorylation of TH on Ser40 increased its affinity towards the yeast 14-3-3 isoforms BMH1/BMH2 and sheep brain 14-3-3, but not for 14-3-3 zeta. On phosphorylation of both Ser19 and Ser40, binding to the 14-3-3 zeta isoform also occurred, and the binding affinity to BMH1 and sheep brain 14-3-3 increased. Both phosphoserine-specific antibodies directed against the 10 amino acids surrounding Ser19 or Ser40 of TH, and the phosphorylated peptides themselves, inhibited the association between phosphorylated TH and 14-3-3 proteins. This was also found when heparin was added, or after proteolytic removal of the N-terminal 37 amino acids of Ser40-phosphorylated TH. Binding of BMH1 to phosphorylated TH decreased the rate of dephosphorylation by protein phosphatase 2A, but no significant change in enzymatic activity was observed in the presence of BMH1. These findings further support a role for 14-3-3 proteins in the regulation of catecholamine biosynthesis and demonstrate isoform specificity for both TH and 14-3-3 proteins.