Developing a control framework for self-adjusting prosthetic sockets incorporating tissue injury risk estimation and generalized predictive control.

To perform activities of daily living (ADL), people with lower limb amputation depend on the prosthetic socket for stability and proprioceptive feedback. Poorly fitting sockets can cause discomfort, pain, limb tissue injuries, limited device usage, and potential rejection. Semi-passively controlled adjustable socket technologies exist, but these depend upon the user's perception to determine safe interfacial pressure levels. This paper presents a framework for automatic control of an adjustable transtibial prosthetic socket that enables active adaptation of residuum-socket interfacial loading through localized actuators, based on soft tissue injury risk estimation. Using finite element analysis, local interfacial pressure vs. compressive tissue strain relationships were estimated for three discrete anatomical actuator locations, for tissue injury risk assessment within a control structure. Generalized Predictive Control of multiple actuators was implemented to maintain interfacial pressure within estimated safe and functional limits. Controller simulation predicted satisfactory dynamic performance in several scenarios. Actuation rates of 0.06-1.51 kPa/s with 0.67% maximum overshoot, and 0.75-1.58 kPa/s were estimated for continuous walking, and for a demonstrative loading sequence of ADL, respectively. The developed platform could be useful for extending recent efforts in adjustable lower limb prosthetic socket design, particularly for individuals with residuum sensory impairment.

Multiscale, multidomain analysis of microvascular flow dynamics.

NEW FINDINGS: What is the topic of this review? We describe a range of techniques in the time, frequency and information domains and their application alone and together for the analysis of blood flux signals acquired using laser Doppler fluximetry. What advances does it highlight? This review highlights the idea of using quantitative measures in different domains and scales to gain a better mechanistic understanding of the complex behaviours in the microcirculation. ABSTRACT: To date, time- and frequency-domain metrics of signals acquired through laser Doppler fluximetry have been unable to provide consistent and robust measures of the changes that occur in the microcirculation in healthy individuals at rest or in response to a provocation, or in patient cohorts. Recent studies have shown that in many disease states, such as metabolic and cardiovascular disease, there appears to be a reduction in the adaptive capabilities of the microvascular network and a consequent reduction in physiological information content. Here, we introduce non-linear measures for assessing the information content of fluximetry signals and demonstrate how they can yield deeper understanding of network behaviour. In addition, we show how these methods may be adapted to accommodate the multiple time scales modulating blood flow and how they can be used in combination with time- and frequency-domain metrics to discriminate more effectively between the different mechanistic influences on network properties.

Metabolic regulation during constant moderate physical exertion in extreme conditions in Type 1 diabetes.

BACKGROUND: Constant moderate intensity physical exertion in humid environments at altitude poses a considerable challenge to maintaining euglycaemia with Type 1 diabetes. Blood glucose concentrations and energy expenditure were continuously recorded in a person trekking at altitude in a tropical climate to quantify changes in glucose concentrations in relation to energy expenditure. CASE REPORT: Blood glucose concentrations and energy expenditure were continuously monitored with a Guardian® real-time continuous glucose monitoring system (CGMS) and a SenseWear® Pro3 armband (BodyMedia Inc., USA), in a 27-year-old woman with Type 1 diabetes, during her climb up Mount Kinabalu in Borneo (c. 4095 m). Comparative control data from the same person was collected in the UK (temperate climate at sea level) and Singapore (tropical climate at sea level). Maximum physical effort during the climb was < 60% VO(2MAX) (maximal oxygen consumption). Mean daily calorific intakes were 2300 kcal (UK), 2370 kcal (Singapore) and 2274 kcal (Mount Kinabalu), and mean daily insulin doses were 54 U (UK), 40 U (Singapore) and 47 U (Mount Kinabalu). Despite markedly increased energy expenditure during the climb [4202 kcal (Mount Kinabalu) vs. 2948 kcal (UK) and 2662 kcal (Singapore)], mean blood glucose was considerably higher during the trek up Mount Kinabalu [13.2 ± 5.9 mmol/l, vs. 7.9 ± 3.8 mmol/l (UK) and 8.6 ± 4.0 mmol/l (Singapore)]. CONCLUSION: Marked unexpected hyperglycaemia occurred while trekking on Mount Kinabalu, despite similar calorie consumption and insulin doses to control conditions. Because of the risk of unexpected hyperglycaemia in these conditions, we recommend that patients embarking on similar activity holidays undertake frequent blood glucose monitoring.

Diastolic function is strongly and independently associated with cardiorespiratory fitness in central obesity.

Cardiorespiratory fitness [maximal O2 consumption (VO2max)] is an independent risk factor for type 2 diabetes; but in individuals at risk, factors influencing VO2max are poorly understood. We tested the hypothesis that VO2max is associated with diastolic function [subendocardial variability ratio (SEVR), %], as diastolic function influences myocardial perfusion. We studied 47 men and women with central obesity without diabetes. We measured fitness (VO2max) by treadmill testing and diastolic function (SEVR%) by pulse-wave analysis. We measured other factors influencing this relationship: insulin sensitivity [whole body glucose uptake-to-insulin concentration ratio (M/I)] by hyperinsulinemic euglycemic clamp, fatness by MR imaging and dual-energy X-ray absorptiometry, physical activity energy expenditure (metabolic equivalents of tasks) by the Sensewear Pro2 device, and muscle microvascular exchange capacity (capillary filtration coefficient) by venous plethysmography. Mean age of the subjects was 51+/-9 (SD) yr. VO2max was associated with SEVR% (r=0.50, P=0.001), fatness (r=-0.39, P=0.008), and HbA1c (r=-0.35, P=0.018), but not with whole body glucose uptake-to-insulin concentration ratio, metabolic equivalents of tasks, or capillary filtration coefficient. In regression modeling with age, sex, fatness, and SEVR% as explanatory variables, only age, sex, and SEVR% were independently associated with VO2max (SEVR%--standardized B coefficient=0.37, 95% confidence interval=0.003-0.18, P=0.007). This model identified 46% of the variance in VO2max (R2=0.46, P=0.0001). There was a strong, independent association between VO2max and a measure of diastolic function in sedentary individuals with central obesity.

Skeletal muscle microvascular exchange capacity is associated with hyperglycaemia in subjects with central obesity.

AIMS: Poor glycaemic control is associated with increased risk of microvascular disease in various organs including the eye and kidney, but the relationship between glycated haemoglobin (HbA(1c)) and microvascular function in skeletal muscle has not been described. We tested the association between HbA(1c) and a measure of microvascular exchange capacity (K(f)) in skeletal muscle in people with central obesity at risk of developing Type 2 diabetes. METHODS: Microvascular function was measured in 28 women and 19 men [mean (+/- sd) age 51 +/- 9 years] with central obesity who did not have diabetes. We estimated insulin sensitivity by hyperinsulinaemic-euglycaemic clamp, visceral and total fatness by magnetic resonance imaging, fitness (VO(2) max by treadmill testing), physical activity energy expenditure [metabolic equivalents of tasks (METS) by use of the SenseWear Pro armband] and skeletal muscle microvascular exchange capacity (K(f)) by venous occlusion plethysmography. RESULTS: In regression modelling, age, sex and fasting plasma glucose accounted for 30.5% of the variance in HbA(1c) (r(2) = 0.31, P = 0.001). Adding K(f) to this model explained an additional 26.5% of the variance in HbA(1c) (r(2) = 0.57, P = 0.0001 and K(f) was strongly and independently associated with HbA(1c) (standardized B coefficient -0.45 (95% confidence interval -0.19, -0.06), P = 0.001). CONCLUSIONS: We found a strong negative independent association between a measure of skeletal muscle microvascular exchange capacity (K(f)) and HbA(1c). K(f) was associated with almost as much of the variance in HbA(1c) as fasting plasma glucose.

Healthy habits: efficacy of simple advice on weight control based on a habit-formation model.

OBJECTIVE: To evaluate the efficacy of a simple weight loss intervention, based on principles of habit formation. DESIGN: An exploratory trial in which overweight and obese adults were randomized either to a habit-based intervention condition (with two subgroups given weekly vs monthly weighing; n=33, n=36) or to a waiting-list control condition (n=35) over 8 weeks. Intervention participants were followed up for 8 months. PARTICIPANTS: A total of 104 adults (35 men, 69 women) with an average BMI of 30.9 kg m(-2). INTERVENTION: Intervention participants were given a leaflet containing advice on habit formation and simple recommendations for eating and activity behaviours promoting negative energy balance, together with a self-monitoring checklist. MAIN OUTCOME MEASURES: Weight change over 8 weeks in the intervention condition compared with the control condition and weight loss maintenance over 32 weeks in the intervention condition. RESULTS: At 8 weeks, people in the intervention condition had lost significantly more weight (mean=2.0 kg) than those in the control condition (0.4 kg), with no difference between weekly and monthly weighing subgroups. At 32 weeks, those who remained in the study had lost an average of 3.8 kg, with 54% losing 5% or more of their body weight. An intention-to-treat analysis (based on last-observation-carried-forward) reduced this to 2.6 kg, with 26% achieving a 5% weight loss. CONCLUSIONS: This easily disseminable, low-cost, simple intervention produced clinically significant weight loss. In limited resource settings it has potential as a tool for obesity management.

The three mechanisms of intracellular chloride accumulation in vascular smooth muscle of human umbilical and placental arteries.

Recordings of membrane potential (Em) and intracellular [Cl-] ([Cl-]i) were made from the smooth muscle of human umbilical and placental arteries, using double-barrelled, ion-sensitive microelectrodes. In both arteries, [Cl-]i was above equilibrium with Em. In the umbilical artery, [Cl-]i was 33.8+/-0.9 mM (+/-SD, n=19) and Em -54.9+/-1.3 mV and in the placental artery respectively 35.1+/-0.7 mM (n=17) and -50.6+/-0.9 mV. In both arteries, [Cl-]i was reduced and Em hyperpolarised significantly by successive additions of 100 microM 4,4'-diisothiocyanatodihydrostilbene-2,2'-disulphonic acid (DIDS), 10 microM bumetanide and 1 mM acetazolamide, thus revealing the presence of Cl-/HCO3- exchange, (Na+K+Cl) cotransport and "pump III". In the presence of all three inhibitors, [Cl-]i was in equilibrium with Em. As in earlier studies on rat arterial smooth and cardiac muscle, pump III was unaffected by DIDS, bumetanide, metolazone and the removal of Na+, partly inhibited by chlorothiazide and fully inhibited by ethacrynic acid. The results are discussed in terms of the possibility that of chloride accumulating systems may regulate vasomotor tone in the foetoplacental unit.

Chloride in smooth muscle.

Interest in the functions of intracellular chloride expanded about twenty years ago but mostly this referred to tissues other than smooth muscle. On the other hand, accumulation of chloride above equilibrium seems to have been recognised more readily in smooth muscle. Experimental data is used to show by calculation that the Donnan equilibrium cannot account for the chloride distribution in smooth muscle but it can in skeletal muscle. The evidence that chloride is normally above equilibrium in smooth muscle is discussed and comparisons are made with skeletal and cardiac muscle. The accent is on vascular smooth muscle and the mechanisms of accumulation and dissipation. The three mechanisms by which chloride can be accumulated are described with some emphasis on calculating the driving forces, where this is possible. The mechanisms are chloride/bicarbonate exchange, (Na+K+Cl) cotransport and a novel entity, "pump III", known only from own work. Their contributions to chloride accumulation vary and appear to be characteristic of individual smooth muscles. Thus, (Na+K+Cl) always drives chloride inwards, chloride/bicarbonate exchange is always present but does not always do it and "pump III" is not universal. Three quite different biophysical approaches to assessing chloride permeability are considered and the calculations underlying them are worked out fully. Comparisons with other tissues are made to illustrate that low chloride permeability is a feature of smooth muscle. Some of the functions of the high intracellular chloride concentrations are considered. This includes calculations to illustrate its depolarising influence on the membrane potential, a concept which, experience tells us, some people find confusing. The major topic is the role of chloride in the regulation of smooth muscle contractility. Whilst there is strong evidence that the opening of the calcium-dependent chloride channel leads to depolarisation, calcium entry and contraction in some smooth muscles, it appears that chloride serves a different function in others. Thus, although activation and inhibition of (Na+K+Cl) cotransport is associated with contraction and relaxation respectively, the converse association of inhibition and contraction has been seen. Nevertheless, inhibition of chloride/bicarbonate exchange and "pump III" and stimulation of (K+Cl) cotransport can all cause relaxation and this suggests that chloride is always involved in the contraction of smooth muscle. The evidence that (Na+K+Cl) cotransport more active in experimental hypertension is discussed. This is a common but not universal observation. The information comes almost exclusively from work on cultured cells, usually from rat aorta. Nevertheless, work on smooth muscle freshly isolated from hypertensive rats confirms that (Na+K+Cl) cotransport is activated in hypertension but there are several other differences, of which the depolarisation of the membrane potential may be the most important.Finally, a simple calculation is made which indicates as much as 40% of the energy put into the smooth muscle cell membrane by the sodium pump is necessary to drive (Na+K+Cl) cotransport. Notwithstanding the approximations in this calculation, this suggests that chloride accumulation is energetically expensive. Presumably, this is related to the apparently universal role of chloride in contraction.

Increased (Na+K+Cl) cotransport in rat arterial smooth muscle in deoxycorticosterone (DOCA)/salt-induced hypertension.

The inhibition by loop diuretics of K efflux (tracer (86)Rb) from the rat femoral arterial smooth muscle was measured in normotension and in DOCA-salt hypertension. The sensitivity sequence (bumetanide > piretanide > furosemide) was the characteristic pharmacological profile of (Na+K+Cl) cotransport. In hypertension, cotransport activity was 46% greater than in normotension and the sensitivity to loop diuretics was threefold less. Intracellular ¿K and the Na, K and Cl permeability ratios and electrogenic Na pump activity were assessed electrophysiologically in normotension and hypertension. ¿K(i) was lower in hypertension (173 mM) than normotension (198 mM) but the other parameters (P(Na/Cl) = 0.14, P(Cl)/P(K) = 0.19 and electrogenic pump = -8.3 mV in normotension) were not significantly different. Ionic permeabilities to Na, K and Cl were significantly lower in hypertension than normotension. Plasma ¿Na, but not ¿K, was higher in hypertension than normotension. The conclusion is that increased activation of (Na+K+Cl) cotransport in hypertension plays a major role in the elevation of ¿Cl(i) and depolarisation of the membrane potential in vascular smooth muscle in DOCA-salt hypertension. The role of (Na+K+Cl) cotransport in vascular smooth muscle in this model of hypertension is discussed in relation to ¿Cl(i), depolarisation of the membrane potential and contraction and in relation to cell growth.

Activation of two inward chloride transport systems in rat femoral arterial smooth muscle in deoxycorticosterone acetate/salt hypertension.

1. Intracellular [Cl-] ([Cl-]i) was measured with ion-selective microelectrodes in rat femoral arterial smooth muscle in normotensive controls and after the induction of deoxycorticosterone acetate/salt hypertension. 2. Linear regression of [Cl-]i and time after the induction of hypertension showed good correlation (r = 0.96) for 5-6 weeks, as [Cl-]i increased from 30 +/- 1 mmol/l (mean +/- SD, n = 16), to 49 +/- 2 mmol/l (n = 9, P < 0.0001). 3. Arterial systolic blood pressure also increased linearly (r = 0.97) for 5-6 weeks as hypertension developed from 122 +/- 1 mmHg (n = 20) to 187 +/- 7 mmHg (n = 14): there was consequently a linear relationship between [Cl-]i and arterial systolic blood pressure (r = 0.96). 4. The increase in [Cl-]i was partly because Na(+)-K(+)-Cl- co-transport activity, estimated from the fall in [Cl-]i caused by bumetanide, was greater in hypertension (18 mmol/l) than in normotension (10 mmol/l). This finding, and the depolarization of the membrane potential in hypertension (-56 +/- 3 mV compared with -64 +/- 4 mV in normotension; P < 0.0001), confirms previous studies. 5. The increase in [Cl-]i was also partly due to greater activity of an Na(+)- and HCO3(-)-independent, acetazolamide-sensitive inward Cl- transport system; thus acetazolamide reduced [Cl-]i by 7 mmol/l in normotension and by 16 mmol/l in hypertension. 6. In Cl(-)-free media, the membrane potential in normotension (-59 +/- 5 mV) was not significantly different from that in hypertension (-60 +/- 4 mV). 7. The role of [Cl-]i in the depolarization of the membrane potential in hypertension is discussed.

Sodium-independent inward chloride pumping in rat cardiac ventricular cells.

The intracellular Cl concentration ([Cl]i) in rat cardiac ventricular muscle, measured with double-barreled microelectrodes in vitro, was 21.3 +/- 1.5 (SD) mM [number of observations (n) = 46]. With the Na-K-Cl cotransport inhibitor bumetanide (10 microM), it fell to 13.4 +/- 1.4 mM (n = 27), and with 1 mM acetazolamide, it fell further, to 7.2 +/- 1.5 mM (n = 5), close to equilibrium with the membrane potential. In the absence of Na, [Cl]i was 15.9 +/- 1.4 mM (n = 8), and with 1 mM acetazolamide, it fell to 6.5 +/- 0.6 mM (n = 4), again close to equilibrium. The bumetanide- and Na-insensitive components of inward Cl pumping were inhibited by chlorothiazide and ethacrynic acid but were unaffected by the Na-Cl cotransport inhibitor metolazone. There was inhibition of Na-K-Cl cotransport by chlorothiazide = acetazolamide > metolazone. The anion exchange inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid and HCO3 had no effect on [Cl]i in any condition. Thus Cl accumulation in the rat ventricle is fully accounted for by two systems, namely, Na-K-Cl cotransport and an Na-independent, possibly primary active, process.

Stimulation of intracellular chloride accumulation by noradrenaline and hence potentiation of its depolarization of rat arterial smooth muscle in vitro.

1. Double-barrelled ion-selective microelectrodes were used to examine the effects of exogenous noradrenaline upon the membrane potential (Em) and intracellular chloride concentration ([Cl]i) of arterial smooth muscle from the saphenous branch of the femoral artery of the rat. 2. After treatment with 0.6 mM 6-hydroxydopamine (to functionally denervate the tissue), exogenous noradrenaline (5 nM) caused repeatable depolarization of Em from -63.7 +/- 2.4 mV (s.d., n = 18) to -53.8 +/- 3.4 mV (P < 0.0001) and increases in [Cl]i from 31.0 +/- 0.5 mM to 42.5 +/- 2.2 mM (P < 0.0001). 3. In the presence of 10 microM bumetanide (an inhibitor of (Na-K-Cl) cotransport), 5 nM noradrenaline caused a depolarization of Em of 3.0 +/- 3.2 mV, and a rise in [Cl]i of 4.5 +/- 2.5 mM. 4. In the presence of bumetanide and 1 mM acetazolamide (used as an inhibitor of a Na-independent inward Cl pump), noradrenaline had no effect on Em or [Cl]i. 5. In the absence of extracellular chloride, the rise in apparent [Cl]i in response to 5 nM noradrenaline was abolished but there was a depolarization of 2.0 +/- 3.9 mV. 6. These results are consistent with the stimulation of (Na-K-Cl) cotransport and a Na-independent Cl pump by exogenous noradrenaline and with the consequent increase in [Cl]i and shift in ECl potentiating the depolarization caused by noradrenaline. The possibility that modulation of [Cl]i may be a general mechanism of Em regulation is discussed.

Chronotropic action of Na+, K+, Cl- cotransport inhibition in the isolated rat heart.

The chronotropic actions of Na+, K+, Cl- cotransport were investigated by studying the effects of the loop diuretics bumetanide and furosemide, specific inhibitors of the cotransporter, on an isolated rat sino-atrial node preparation. Application of bumetanide decreased the cycle length from 0.334 s (+/- 0.087 S.D.) to 0.279 s (+/- 0.083, n = 16, P = 6.5 x 10(-6)) in Hepes-buffered physiological salt solution (PSS). Similar decreases were recorded in bicarbonate-buffered PSS. Chloride channel blockers indicate that the tachycardia evoked by loop diuretics is not due their blocking of chloride channels. Thus, 4,4'-dinitrostilbene-2,2'-disulphonic acid (DNDS) and 5-nitro-2-(3-phenylpropylamino) benzoic acid (NPPB) had a negative chronotropic action and 2-[(2-cyclopentyl-6,7-dichloro-2,3-dihydro-2-methyl-1-oxo-1H-inden -5-yl) oxy] acetic acid (IAA-94) produced no change in cycle length. Pharmacological manoeuvres indicate that the positive chronotropic action of loop diuretics is associated with catecholamine release. The positive chronotropic action of bumetanide was inhibited by the beta-adrenoceptor antagonists, propranolol and atenolol, but was unaffected by atropine.

An acetazolamide-sensitive inward chloride pump in vascular smooth muscle.

In smooth muscle cells, which have a small but significant chloride ion permeability, Cl:HCO3 exchange and (Na+K+Cl) cotransport are known to act as inwardly directed chloride pumps. However, even allowing for overestimation of [Cl]i due to intracellular interference with the Cl-recording electrodes, there remained a residual accumulation of chloride in rat arterial smooth muscle under conditions in which neither of these processes is operative. Manipulation of [Cl]i and Cl permeability using pharmacological agents whilst monitoring both Em and [Cl]i showed that an acetazolamide-sensitive mechanism, perhaps a Cl-dependent ATPase, is the third pathway for Cl accumulation.

Accumulation of intracellular chloride by (Na-K-Cl) co-transport in rat arterial smooth muscle is enhanced in deoxycorticosterone acetate (DOCA)/salt hypertension.

The accumulation of intracellular chloride above equilibrium by (Na-K-Cl) co-transport has been demonstrated in a variety of tissues, most recently in guinea-pig vas deferens. The depolarizing influence of the co-transporter on the membrane potential of arterial smooth muscle cells has been demonstrated in rat femoral artery. The inference is that the depolarisation of membrane potential seen in deoxycorticosterone acetate (DOCA)/salt hypertension is the result of increased chloride accumulation via (Na-K-Cl) co-transport, which is enhanced in hypertension. The questions addressed here are (i) whether chloride is accumulated above equilibrium in saphenous arterial smooth muscle cells from normotensive animals, and (ii) whether the accumulation is more pronounced in association with DOCA/salt hypertension. In arterial smooth muscle from DOCA/salt hypertensive animals, [Cl-]i was significantly elevated (P < 2 x 10(-9)) in comparison to arterial smooth muscle from normotensive control animals. The loop diuretic bumetanide caused a reversible hyperpolarization of Em and a fall in [Cl-]i and these effects were enhanced in hypertension. These results are consistent with an increase in the activity of the (Na-K-Cl) co-transporter in rat femoral arterial smooth muscle in DOCA/salt hypertension. A preliminary report of this work has been published.

Uptake of zinc by human placental microvillus border membranes and characterization of the effects of cadmium on this process.

The uptake of Zinc (Zn) by microvillus border membrane vesicles formed from the trophoblast of term human placentae is markedly increased over brief periods of incubation with much slower increases persisting for up to 2 h of incubation. Zinc is both bound to membrane components and transported into intravesicular osmotically active space. Uptake is saturable, temperature dependent from 4 to 37 degrees C with a Q10 of 1.7, and is inhibited by the sulphydryl agent DTNB. About 20 per cent of the uptake is susceptible to inhibition by Cadmium (Cd) at concentrations from 5 to 50 microM, a significant part of the action of this metal being on the transmembrane component of Zn uptake.

Evidence for volume regulatory increase by Na-K-Cl co-transport in rat arterial smooth muscle.

A component of K efflux (86Rb as tracer) from rat femoral arterial smooth muscle was blocked by loop diuretics. Following hypertonic challenge with 100 mM-sucrose, K efflux increased but not in the presence of loop diuretics. This is taken as evidence for a volume regulatory increase mediated by Na-K-Cl co-transport.

Comparison of the electrical properties of arterial smooth muscle in normotensive rats and rats with deoxycorticosterone acetate-salt-induced hypertension: possible involvement of (Na(+)-K(+)-Cl-) co-transport.

1. Hypertension was induced in male Sprague-Dawley rats by left unilateral nephrectomy and deoxycorticosterone acetate--salt administration. After 5 weeks, arterial systolic blood pressure was significantly elevated in these animals (191.5 +/- 6.0 mmHg, mean +/- SD, n = 17) compared with age-matched, unoperated control animals (134.0 +/- 4.2 mmHg, n = 8, P less than 0.001). 2. The membrane potential of femoral artery vascular smooth muscle measured in vitro was -55.1 +/- 6.3 mV (mean +/- SD, n = 154) for normotensive and -50.8 +/- 5.7 mV (n = 82) for hypertensive animals. The difference in membrane potential was significant (P less than 0.001). 3. The relationship between the log of the extracellular K+ concentration and membrane potential was nonlinear over the extracellular K+ concentration range 2.5-20 mmol/l, and showed a small positive shift with hypertension. 4. Tenfold reductions in the extracellular concentrations of Na+ or Cl- resulted in a membrane potential hyperpolarization in vascular smooth muscle from normotensive animals (4.9 +/- 2.0 mV, n = 13 and 12.1 +/- 1.3 mV, mean +/- SD, n = 14, respectively). In vascular smooth muscle from hypertensive animals, the hyperpolarization in low-Na+ media was significantly increased to 12.2 +/- 2.6 mV (mean +/- SD, n = 5), but that in low-Cl- media was unaffected (2.7 +/- 1.6 mV, n = 6). 5. The loop diuretic, bumetanide (10 mumol/l), hyperpolarized the membrane potential in vascular smooth muscle from both normotensive and hypertensive rats, but not in low-Na+ or low-Cl- media.(ABSTRACT TRUNCATED AT 250 WORDS)