Enhancement Of Na Research Articles

Muscarinic activation causes biphasic inotropic response and decreases cellular Na+ activity in canine cardiac Purkinje fibers.

In this study, the effects of carbachol (CCh) on twitch tension, intracellular Na+ activity (aiNa), and action potential were simultaneously measured in canine cardiac Purkinje fibers in order to examine the regulation of inotropy through muscarinic receptors and its relation to aiNa. In fibers driven at 1 Hz, CCh (10 microM) initially and transiently decreased and then increased the twitch tension by 36 +/- 8%. The action potential showed a significant elevation of the plateau and a significant shortening of the duration at 90% repolarization (APD90), from 403 +/- 7 to 389 +/- 7 ms. The aiNa decreased from 7.4 +/- 0.4 to 6.7 +/- 0.3 mM (n = 23, p < 0.05). Atropine (1 microM) decreased the twitch tension by 21 +/- 6% (n = 7, p < 0.05) without significant effects on the action potential and aiNa, and inhibited the effects of CCh. Cs+ (20 mM) increased the plateau height and APD90, enhanced the twitch tension by 66 +/- 24%, but decreased aiNa from 7.3 +/- 0.3 to 6.3 +/- 0.4 mM (n = 6, p < 0.05). In the presence of 20 mM Cs+, some fibers generated slow responses. The addition of 10 microM CCh further increased the twitch tension and APD90, and decreased aiNa from 6.3 +/- 0.4 to 5.3 +/- 0.3 mM. Ouabain (0.3 microM) increased the twitch tension and aiNa, and inhibited the CCh-induced decrease of aiNa. In the presence of ouabain, 20 mM Cs+ depolarized the fiber and generated slow responses with a decreased aiNa. The addition of 10 microM CCh enhanced the slow action potential, and increased aiNa although there was a transient decrease during early exposure. These results suggest that activation of muscarinic receptors in canine Purkinje fibers results in an enhancement of the Na+-K+ pump activity and a biphasic inotropic response, probably via different receptor subtypes. The inhibitory effect, most likely through M2 receptors, is associated with the activation of K+ channels. The stimulatory effect, on the other hand, is probably due to the action on the M1 receptors, resulting in increases in Ca2+ currents.

Heat shock-induced protection and enhancement of Na+-glucose cotransport by LLC-PK1 monolayers.

Monolayers of the porcine-derived renal epithelial cell line, LLC-PK1, were used to characterize the effects of heat stress on Na+-glucose cotransport. Transepithelial current dependent on 5 mM glucose (I(Glc)), phloridzin-sensitive current (I(phz)), and total transepithelial current (I(total)) were measured as indicators of Na+-glucose cotransport. Severe heat shock (SHS; 45 degrees C for 1 h, then 37 degrees C for measurements) decreased transepithelial electrical resistance (TER), I(Glc), I(phz), and I(total) 50-70%. Mild heat shock (MHS; 42 degrees C for 3 h, then 37 degrees C for 12 h) induced accumulation of 72-kDa heat shock protein (HSP-72), decreased damage to TER from SHS, and prevented damage to I(Glc), I(phz), and I(total). Kinetic analysis showed that SHS damaged and MHS protected total Na+-glucose transport capacity (Vmax of I(Glc)). MHS alone increased TER (50%), I(Glc) (20%), I(total) (20%), and Vmax of I(Glc) (25%). On enhancement of the Na+ gradient by depletion of intracellular Na+, MHS increased I(Glc) 50% and had no effect on transepithelial Na+-dependent sulfate reabsorptive flux measured concurrently or in Na+-replete tissues. These effects of MHS were not reflected in effects on cell survival or luminal membrane surface area as indicated by lactate dehydrogenase or alkaline phosphatase release. In conclusion, HSP-72-inducing heat treatment both protected and enhanced Na+-glucose cotransport independently of the luminal membrane Na+ gradient and selectively with respect to effects on TER, reabsorptive sulfate transport, cell survival, and luminal membrane surface area.

Plasma triglycerides and red cell ion transport alterations in genetically hypertensive rats.

Ion transport abnormalities in essential hypertension are often associated with concomitant changes of lipid metabolism, but this information is missing in rats with genetic hypertension. We therefore studied the alterations of red cell Na+ and K+ transport and their relationship to blood pressure and plasma lipids (cholesterol and triglycerides) in Prague hereditary hypertriglyceridemic (HTG) rats, Lyon hypertensive (LH) rats, and HTG x Lewis F2 hybrids. In both hypertensive models and F2 hybrids, red cell Na+ content (Na+(i)) was positively related to plasma triglycerides but not to plasma cholesterol levels. Na+(i) elevation was more pronounced in HTG than in LH rats, probably due to higher plasma triglycerides in the former strain. The two hypertensive strains differed in bumetanide-sensitive Na+ transport, which was augmented in HTG rats with low plasma cholesterol but suppressed in LH rats characterized by high cholesterol levels. In the two genetic models, there was a positive association of blood pressure with Na+ leak, and this was also confirmed by the cosegregation of these parameters in F2 hybrids. We conclude that the enhancement of Na+ leak represents the major ion transport abnormality in rats with genetic hypertension. The alterations in plasma lipids are important determinants of abnormal red cell ion transport in hypertensive models studied. Although the detailed mechanism of their participation in ion transport regulation is still not completely understood, triglyceride-dependent changes in membrane microviscosity seem to be responsible for the modulation of particular ion transport pathways.

Na+ channel modulation and force-frequency relationship in human myocardium.

The enhancement of force of contraction (FOC) following increasing frequencies of stimulation is an important mechanism of positive inotropy in human myocardium. The present study aimed to investigate the influence of alterations in Na+ influx on FFR in human myocardium. Isometric FOC of electrically stimulated right auricular trabeculae (AUT, n = 12) from human non-failing hearts (n = 8) was measured at different stimulation rates (0.5-3 Hz) under control conditions, after increasing Na+ influx by the addition of (+/-)BDF 9148 (BDF, 3 mumol l-1) and after decreasing Na+ influx by the addition of lidocaine (LIDO, 10 mumol l-1). Additionally, the rate dependent changes in diastolic tension (DT) were measured in all experiments. Under control conditions FOC increased with increasing frequencies of stimulation. The rate at which maximal FOC was observed (SFmax) was 2.0 +/- 0.2 Hz and maximal increase in FOC (PIEmax) by increasing frequency of stimulation was +1.5 +/- 0.5 mN. After increase of Na+ influx by BDF (3 mumol l-1) SFmax was decreased to 0.8 +/- 0.1 Hz (p < 0.05 versus control) and PIEmax was +0.1 +/- 0.3 mN (p < 0.05). When Na+ influx was diminished by LIDO (10 mumol l-1) SFmax and PIEmax were increased compared to control (2.4 +/- 0.1 Hz and +4.1 +/- 0.9 mN, p < 0.05 versus control). The diastolic tension (DT) of AUT at 3 Hz was not changed at higher rates in the control group and after application of LIDO (10 mumol l-1), whereas after enhancement of Na+ influx by BDF there was an increase in DT of +0.7 +/- 0.2 at 3 Hz (p < 0.05 versus control and LIDO). An enhanced Na+ influx leads to a decrease in the optimal frequency and to a smaller force potentiation by higher stimulation rates which could be at least partly due to incomplete relaxation at higher frequencies, whereas a reduced Na+ influx is followed by opposite alterations. It is concluded that besides Ca2+ handling also Na+ influx and Na+ homeostasis might determine the frequency-induced force potentiation in human myocardium. Thus, the negative FFR in diseased human myocardium might result from changes in cellular Ca2+ or Na+ regulatory sites.

Abundance anomalies in globular clusters

The galactic globular clusters presumably formed rapidly as high density concentrations at the centres of extensive star forming regions and the natural expectation is that they will be chemically homogeneous. In general, this is confirmed by observation — most globular clusters are extremely homogeneous with regard to most elements. (There are two exceptions: ω Cen and M22 both have intrinsic internal abundance ranges. While of considerable interest, e.g. Norris et al. 1996, this type of “abundance anomaly” will not be discussed here). But we have known for more than twenty years that the surface abundances, which are what we observe, of elements such as C, N and O can vary substantially from red giant to red giant within an individual globular cluster. Indeed it has become clear that “abundance anomalies” of this type are common in the galactic globular cluster population. Briefly, the observed anomalies can be summarized as: (i) the “anomalous” stars are depleted in C and enhanced in N. Depletions of O also often accompany the depletions in C. (ii) The C, N and O variations are usually accompanied by enhancements of Na and Al and when the O depletion and the Al enhancement are both large, Mg is depleted. No other elements, including r- and s-process indicators, vary.

Evidence for functional relevance of an enhanced expression of the Na(+)-Ca2+ exchanger in failing human myocardium.

The present study aimed at investigating the expression of the Na(+)-Ca2+ exchanger and its functional role in human failing myocardium. Na(+)-Ca2+ exchanger mRNA and protein levels were examined in nonfailing (NF, n = 8) and failing human myocardium (New York Heart Association functional class IV) with idiopathic dilated cardiomyopathy (DCM, n = 8) or ischemic heart disease (ICM, n = 6). The inotropic effect of the Na+ channel activator BDF 9148 was determined in electrically driven left ventricular papillary muscle strip preparations (1 Hz, 37 degrees C) from nonfailing (n = 8) and failing (n = 8) human hearts. Na(+)-Ca2+ exchanger mRNA levels were significantly increased, by 79% (P < .001) in DCM and by 58% (P < .01) in ICM compared with NF; protein levels increased by 36% (P < .001) and by 20% (P < .05), respectively. BDF 9148 increased the force of contraction concentration dependently, with a similar maximal effect in NYHA class IV and NF, but was more potent in NYHA class IV as demonstrated by a significantly smaller (P < .01) EC50 value (NYHA class IV, 0.18 [0.16 to 0.22] mumol/L; NF, 1.65 [1.3 to 3.0] mumol/L). In NYHA class IV, BDF 9148 (0.1 mumol/L) restored the positive force-frequency relationship and reduced the frequency-dependent increase in diastolic tension in relation to force of contraction. The increased expression of the Na(+)-Ca2+ exchanger is a possible explanation for the increased inotropic potency of the Na+ channel activator BDF 9148 in failing human myocardium. The increase in exchanger molecules could be of functional relevance for the modulation of cardiac contractility by agents that increase the intracellular Na+ concentration. Enhancement of Na(+)-Ca2+ exchanger activity might be a powerful mechanism for increasing cardiac contractility in chronic heart failure.

Free radicals enhance Na+/Ca2+ exchange in ventricular myocytes

Oxygen-derived free radicals (OFR) have been implicated in the pathogenesis of intracellular Ca2+ overload and the arrhythmias that characterize cardiac reperfusion. These arrhythmias may in large part be due to activation of the pathological transient inward current (ITI). However, the identity of the ITI generated by OFR is uncertain. We previously found that H2O2, an OFR-generating compound, markedly stimulated the ITI elicited by brief caffeine pulses in patch-clamped guinea pig ventricular myocytes. In the present study, using patch-clamped rabbit ventricular myocytes loaded with the Ca(2+)-sensitive indicator fura 2, we have further characterized this ITI and have identified its major component to be Na+/Ca2+ exchange based on its dependence on extracellular Na+ and sarcoplasmic reticulum Ca2+ release, its sensitivity to Ni2+, and the effects of its inhibition on relaxation. The effect on ITI was not unique to H2O2, because another free radical-generating system, xanthine + xanthine oxidase, produced a similar response. We hypothesize that enhancement of Na+/Ca2+ exchange by OFR during reperfusion, when intracellular Na+ is elevated, may promote intracellular Ca2+ overload and triggered arrhythmias.

Na+/H+ exchange in hypertension and in diabetes mellitus--facts and hypotheses.

An enhancement of Na+/H+ exchange (NHE) in blood cells of selected patients with essential hypertension and with diabetic nephropathy has been described by various investigators. Recent studies have shown that enhanced NHE activity persists in immortalized lymphoblasts from these patients after prolonged cell culture and, thus, appears to be under genetic control. Available evidence strongly argues against a mutation in the encoding gene or an overexpression of the NHE. Immortalized cells from hypertensive patients with enhanced NHE activity display two-fold enhanced agonist-induced rises of the cytosolic free Ca2+ concentration and the underlying reason was identified as an increased activation of pertussis toxin (PTX)-sensitive G proteins. The molecular mechanism(s) of this phenomenon have not yet been elucidated. It appears likely that similar changes contribute to the enhanced NHE activity phenotype in diabetic nephropathy, although experimental evidence for this is still lacking. An enhanced activation of PTX-sensitive G proteins could explain many of the hitherto unexplained phenomena in essential hypertension, e.g. inheritance, increased vasoconstriction, hypertrophy of remodeling of arterial blood vessels and the heart, enhanced platelet aggregation etc. In diabetes the same defect could provide the basis for the susceptibility to nephropathy, e.g. by enhancing the deleterious effects of autocrine and paracrine growth factors. Thus, the experimental approach of immortalizing blood cells from patients with essential hypertension and diabetic nephropathy has opened new horizons in the identification of genetically fixed abnormalities in intracellular signal transduction which could contribute to both pathologies and which can now be studied without the confounding influences of the diabetic or hypertensive in vivo milieu.

Enhanced hypo-osmoregulation induced by warm-acclimation in antarctic fish is mediated by increased gill and kidney Na+/K(+)-ATPase activities.

Serum osmolality and serum inorganic ion concentrations were studied in two antarctic fish species, Trematomus bernacchii and T. newnesi, during 5 weeks of acclimation to 4 degrees C and compared with control values for groups acclimated to -1.5 degrees C. Acclimation to 4 degrees C significantly decreased the serum osmolality of both species, thereby increasing their seawater-to-extracellular fluid (ECF) osmotic gradient. The decline in osmolality with acclimation to 4 degrees C was accompanied by significant and rapid losses of Na+ and Cl- during the first 14 days of acclimation and was maintained throughout the study period. At day 35 of acclimation, the lipid composition and microsomal Na+/K(+)-ATPase specific activities at 4 degrees C and 37 degrees C were determined in membranes from gill, kidney, liver and muscle tissues. No warm-induced decrease in fatty acid unsaturation was found in the tissues of either species. In the gills and kidneys of both species, the Na+/K(+)-ATPase activities assayed at 4 degrees C were increased after acclimation to 4 degrees C. The Na+/K(+)-ATPase activities at 37 degrees C increased at the higher acclimation temperature in T. newnesi kidneys and T. bernacchii gills, but in both species there was no compensation to temperature in the liver, regardless of assay temperature. Muscle Na+/K(+)-ATPase activity decreased in response to warm-acclimation in T. bernacchii and T. newnesi assayed at 4 degrees C and 37 degrees C, respectively. During acclimation to 4 degrees C, the discontinuity in the Arrhenius plot of the Na+/K(+)-ATPase activities of T. newnesi gill moved to a lower temperature, whereas that of kidney remained unchanged. The results indicate that acclimation to 4 degrees C induced a decrease in serum osmolality which resulted from the positive compensation of Na+/K(+)-ATPase in osmoregulatory tissues. The enhancement in Na+/K(+)-ATPase activity at 4 degrees C suggests that energy expenditure in antarctic fish may be lessened, in part, by maintaining a reduced seawater-to-ECF osmotic gradient.

Salt-Sensitive Hypertension in Transgenic Mice Overexpressing Na + -Proton Exchanger

Essential hypertension is one of the most common diseases that exacerbate the risk of cardiovascular or cerebrovascular attacks. Although the etiology of essential hypertension remains unclear, recent investigations have revealed that an enhancement of Na(+)-proton (Na(+)-H+) exchange activity is a frequently observed ion transport abnormality in hypertensive patients and animal models. To test the hypothesis that increased Na(+)-H+ exchange causes hypertension, we produced transgenic mice overexpressing Na(+)-H+ exchanger and analyzed their Na+ metabolism and blood pressure. Urinary excretion of water and Na+ was significantly decreased in transgenic mice, and systolic blood pressure was elevated after salt loading. The impaired urinary excretion of Na+ suggested that the Na(+)-H+ exchanger overexpressed in the renal tubules increased reabsorption of Na+, which caused a blood pressure elevation by Na+ retention after excessive salt intake. Our results demonstrate that overexpression of Na(+)-H+ exchanger can be a genetic factor that interacts with excessive salt intake and causes salt-sensitive blood pressure elevation.

Alkali Metal, Alkaline Earth Metal, and Ammonium Ion Selectivities of Dibenzo-16-Crown-5 Compounds with Functional Side Arms in Ion-Selective Electrodes

Potentiometric selectivities of 11 dibenzo-16-crown-5 compounds for alkali metal, alkaline earth metal, and ammonium ions have been determined in solvent polymeric membrane electrodes. The ionophores bear one or two pendent groups on the central carbon of the three-carbon bridge in the polyether ring. Side-arm variation includes OCH[sub 3], OCH[sub 2]CH[sub 2]OCH[sub 3], OCH[sub 2]CO[sub 2]C[sub 2]H[sub 5], OCH[sub 2]C(O)N(C[sub 2]H[sub 5])[sub 2], and OCH[sub 2]C(O)N(C[sub 5]H[sub 11])[sub 2] units. Attachment of a propyl group to the ring carbon that bears an extended, oxygen-containing side arm increases the selectivity for Na[sup +] relative to larger alkali metal and alkaline earth metal cations. For a given side arm, a linear relationship is obtained when the enhancement in Na[sup +] selectivity produced by attachment of a geminal propyl group is plotted against the diameter of the interference ion. Potentiometric responses of the dibenzo-16-crown-5 compounds are rationalized in terms of the crown ether ring size and the oxygen basicity, conformational positioning, and rigidity of the side arm. 22 refs., 3 figs., 2 tabs.

Multiple effects of protein kinase C activators on Na+ currents in mouse neuroblastoma cells

The effects of externally applied different protein kinase C (PKC) activators on Na+ currents in mouse neuroblastoma cells were studied using the perforated-patch (nystatin-based) whole cell voltage clamp technique. Two diacylglycerol-like compounds, OAG (1-oleoyl-2-acetyl-sn-glycerol), and DOG (1-2-dioctanoyl-rac-glycerol) attenuated Na+ currents without affecting the time course of activation or inactivation. The reduction in Na+ current amplitude caused by OAG or DOG was dependent on membrane potential, being more intense at positive voltages. The steady-state activation curve was also unaffected by these substances. However, both OAG and DOG shifted the steady-state inactivation curve of Na+ currents to more hyperpolarized voltages. Surprisingly, phorbol esters did not affect Na+ currents. Cis-unsaturated fatty acids (linoleic, linolenic, and arachidonic) attenuated Na+ currents without modifying the steady-state activation. As with DOG and OAG, cis-unsaturated fatty acids also shifted the steady-state inactivation curve to more negative voltages. Interestingly, inward currents were more effectively attenuated by cis-fatty acids than outward currents. Oleic acid, also a cis-unsaturated fatty acid, enhanced Na+ currents. This enhancement was not accompanied by changes in kinetic or steady-state properties of currents. Enhancement of Na+ currents caused by oleate was voltage dependent, being stronger at negative voltages. The inhibitory or stimulatory effects caused by all PKC activators on Na+ currents were completely prevented by pretreating cells with PKC inhibitors (calphostin C, H7, staurosporine or polymyxin B). By themselves, PKC inhibitors did not affect membrane currents. Trans-unsaturated or saturated fatty acids, which do not activate PKC's, did not modify Na+ currents. Taken together, the experimental results suggest that PKC activation modulates the behavior of Na+ channels by at least three distinct mechanisms. Because qualitatively different results were obtained with different PKC activators, it is not clear how Na+ currents would respond to activation of PKC under physiological conditions.

Renal cortical basolateral Na+/HCO 3 ? cotransporter: I. Partial purification and reconstitution

The renal basolateral Na+/HCO3- cotransporter is the main system responsible for HCO3- transport from proximal tubule cells into the blood. The present study was aimed at purifying and functionally reconstituting the Na+/HCO3- cotransporter protein from rabbit renal cortex. Highly purified rabbit renal cortical basolateral membrane vesicles (hereafter designated as original basolateral membrane), enriched 12-fold in Na-K-ATPase, were solubilized in 2% octylglucoside, and then reconstituted in L-alpha-phosphatidylcholine (proteoliposomes). Na+/HCO3- cotransporter activity was assessed as the difference in 22Na uptake in the presence of HCO3- and gluconate. The activity of the Na+/HCO3- cotransporter was enhanced 18-fold in the solubilized protein reconstituted into proteoliposomes compared to the original basolateral membranes. The reconstituted solubilized purified protein exhibited kinetic properties similar to the cotransporter from original basolateral membranes. In addition, it was like the original cotransporter, inhibited by disulfonic stilbene SITS, and was electrogenic. The catalytic subunit of protein kinase A significantly inhibited Na+/HCO3- cotransporter activity in proteoliposomes. The octylglucoside-solubilized protein was further purified by hydroxylapatite column chromatography, and this resulted in an additional enhancement of Na+/HCO3- cotransporter activity of 80-fold over the original basolateral membranes. The fractions containing the highest activity were further processed by glycerol gradient centrifugation, resulting in a 124- to 300-fold increase in Na+/HCO3- cotransporter activity compared to the original basolateral membranes. SDS-PAGE analysis showed an enhancement of a protein doublet of 56 kD MW in the glycerol gradient fraction. Our results demonstrate that we have partially purified and reconstituted the renal Na+/HCO3- cotransporter and suggest that the 56 kD doublet protein may represent the Na+/HCO3- cotransporter.

Cellular responses to steroids in the enhancement of Na+ transport by rat collecting duct cells in culture. Differences between glucocorticoid and mineralocorticoid hormones.

It has recently been discovered that both mineralocorticoid (MC) and glucocorticoid (GC) hormones can stimulate electrogenic Na+ absorption by mammalian collecting duct cells in culture. In primary cultures of rat inner medullary collecting duct (IMCD) cells, 24-h incubation with either MC or GC agonist stimulates Na+ transport approximately threefold. We have now determined that the effects were not additive, but the time courses were different. As aldosterone is known to stimulate citrate synthase, Na+/K+ ATPase activity, and ouabain binding in cortical collecting duct principal cells, we determined the effects of steroids on these parameters in IMCD cells. MC and GC agonists both produced a small increase in citrate synthase activity. There was no increase in Na+/K+ ATPase activity but specific ouabain binding was increased more than two-fold by either agonist. To determine the role of apical Na+ entry in the steroid-induced effects, the Na+ channel inhibitor, benzamil, was used. Benzamil did not alter the stimulation of citrate synthase activity by either steroid. In contrast, GC stimulation of ouabain binding was prevented by benzamil, whereas MC stimulation was not. We conclude that there are differences in the way that MC and GC hormones produce an increased Na+ transport. Both appear to produce translocation (or activation) of pumps into the basolateral membrane. GC stimulation of pump translocation requires increased Na+ entry whereas MC stimulation does not.

Taurine transport by microvillous membrane vesicles and the perfused cotyledon of the human placenta

Human placental uptake and maternal-to-fetal (M-to-F) net transfer of taurine were evaluated in purified microvillous membrane vesicles (MMV) and the isolated perfused cotyledon. Taurine uptake by MMV was specifically stimulated by an inward Na+ gradient [maximum velocity (Vmax), 24.5 +/- 0.6 pmol.mg-1.30 s-1; Michaelis constant (Km), 6.2 +/- 0.7 microM], with uptake stoichiometry showing approximately 2 Na+/taurine molecule. In the presence or absence of Na+, Cl- did not stimulate uptake. Na(+)-stimulated uptake was enhanced by an outward K+ gradient. beta-Alanine and hypotaurine competitively inhibited uptake of taurine in MMV. Two-way uptake inhibition studies showed no interaction between taurine and non-beta-amino acids. When MMV were preloaded with taurine there was enhancement of Na(+)-stimulated uptake. In the perfused placentas, saturable M-to-F net transfer of taurine was not observed, despite saturation of tissue uptake from the maternal circulation. During 3 h of perfusion, no fetal-to-maternal (F-to-M) gradient formed for taurine; yet, a 2:1 gradient simultaneously occurred for histidine. This study demonstrates that taurine uptake by the microvillous membrane of the human placenta is highly specific, of high affinity, and Na+ coupled.

Taurine transport by microvillous membrane vesicles and the perfused cotyledon of the human placenta

Human placental uptake and maternal-to-fetal (M-to-F) net transfer of taurine were evaluated in purified microvillous membrane vesicles (MMV) and the isolated perfused cotyledon. Taurine uptake by MMV was specifically stimulated by an inward Na+ gradient [maximum velocity (Vmax), 24.5 +/- 0.6 pmol.mg-1.30 s-1; Michaelis constant (Km), 6.2 +/- 0.7 microM], with uptake stoichiometry showing approximately 2 Na+/taurine molecule. In the presence or absence of Na+, Cl- did not stimulate uptake. Na(+)-stimulated uptake was enhanced by an outward K+ gradient. beta-Alanine and hypotaurine competitively inhibited uptake of taurine in MMV. Two-way uptake inhibition studies showed no interaction between taurine and non-beta-amino acids. When MMV were preloaded with taurine there was enhancement of Na(+)-stimulated uptake. In the perfused placentas, saturable M-to-F net transfer of taurine was not observed, despite saturation of tissue uptake from the maternal circulation. During 3 h of perfusion, no fetal-to-maternal (F-to-M) gradient formed for taurine; yet, a 2:1 gradient simultaneously occurred for histidine. This study demonstrates that taurine uptake by the microvillous membrane of the human placenta is highly specific, of high affinity, and Na+ coupled.

Abnormal erythrocyte sodium leak in a subset of essential hypertensive patients

We measured the ouabain- and bumetanide-resistant Na+ efflux in Mg2+-sucrose medium (passive Na+ leak) in erythrocytes from 30 normotensive controls and 72 essential hypertensive patients. The mean values (+/- SEM) of the rate constant of Na+ leak (kpNa) were not significantly different between normotensives and hypertensives. Nevertheless, using the 95% confidence limits of the kpNa (in 10(-3).h-1) in the normotensive group as a cut-off point, 7 (9.7%) essential hypertensives exhibited increased values (58.96 +/- 10.12) when compared with the other 65 patients (23.86 +/- 0.74). revealing increased passive Na+ permeability in the former (leak "+" hypertensives). Na+ fluxes depending on the Na+-K+ pump, outward Na+-K+ cotransport, and Na+-Li+ countertransport were also measured in fresh erythrocytes from the same 72 patients. Three of them (4.2%) exhibited decreased values of ouabain-sensitive Na+ efflux and 6 (8.3%) of bumetanide-sensitive Na+ efflux, while 8 patients (11.1%) showed increased values of Li+-stimulated Na+ efflux and, finally, 48 patients (59.7%) did not present any evident abnormality in these Na+ transport systems. No differences were observed between leak "+" hypertensives and the remaining 65 patients when both basal erythrocyte Na+ content and clinical parameters of hypertension were compared. However, Na+ efflux depending on the outward Na+-K+ cotransport was significantly higher in the leak "+" hypertensive subset (299.43 +/- 43.18 vs 181.52 +/- 10.76 mumol.(l cells.h)-1; P = 0.0078), suggesting a compensatory phenomenon. Enhancement of Na+ permeability detected in 3% to 16% of essential hypertensives may be implicated in the pathogenesis of primary hypertension.

60 ENTEROPATHOGENIC E. COLI (EPEC) DISRUPT SMALL INTESTINAL BRUSH BORDER MEMBRANCE Na+/H* EXCHANGE

Despite a highly specific morphological lesion of the small intestinal brush border membrane, the mechanisms whereby EPEC produce diarrhoea are unknown; mucosal invasion and toxin production are not thought to be important factors. We have therefore studied the effects of the rabbit EPEC strain, RDEC-1, upon Na+/H+ exchange at 60 sec in rabbit small intestinal brush border membrane vesicles (BBMV), following incubation at 37°C with RDEC-1. Before incubation, BBMV showed a 3.4-fold enhancement of Na+ uptake in the presence of an outwardly directed H+ gradient (p<0.0001), indicating intact Na+/H+ exchange. However, after 60 min incubation with RDEC-1; H+-stimulated Na+ uptake was markedly reduced to 43% of original enhancement (p<0.01) but reduced to only 90% in control BBMV (NS).

Thyroid Thermogenesis: Regulation of (Na$$K$)-Adenosine Triphosphatase and Active Na,K Transport

SYNOPSIS. Evidence in support of the hypothesis that T3-induced enhancement of O2 consumption in mammalian target tissues is attributable to a stimulation of energy utilization for active transmembrane Na$ and K$ transport is reviewed. The stimulation of target-tissue Na, K transport-dependent respiration following T3 treatment is associated with enhanced rates of active Na$ efflux and K$ influx as well as with an increase in Na, KATPaseenzymatic activity. The enhancement of Na, K-ATPase activity and enzyme abundance is secondary to a T3-induced stimulation of Na, K-ATPase a and rβ subunit biosynthesis and is probably mediated by increased abundance of specific messenger RNAs coding for the subunits of the enzyme. It is proposed that a coordinate augmentation of Na, KATPase enzymatic activity and enhancement of passive membrane permeability to Na$ and K$ are necessary to maintain the increased rates of active Na, K transport and energy consumption associated with thyroid thermogenesis.

Nutrient leaching from pine needles impacted by acidic cloudwater

In coastal and mountainous environments, fog and cloud droplets are frequently deposited onto terrestrial surfaces; this deposition pathway may account for a large fraction of both moisture and chemical loading. Stratus cloudwater was collected for chemical analyses at Henninger Flats (870 m MSL), a site in the foothills of the San Gabriel Mountains, 25 km northeast of downtown Los Angeles. Concurrent samples of intercepted cloudwater were manually removed, drop-by-drop, from needles on various species of pine trees upon which it had deposited. Samples were analyzed for pH, major cations, and anions. Solute concentrations were substantially higher in samples removed from pine needles compared to the suspended cloudwater. For example, cloudwater concentrations for nitrate and sulfate were measured between 0.25 and 4.5 meq L^(−1) while deposited samples were 0.4 to 90 meq L^(−1). This solute enhancement was due to evaporation following droplet deposition and to nutrient leaching. Nutrient leaching was indicated by (a) a disproportionate increase in the concentrations of cations such as K and Mg (a factor of 2 to 15 enrichment relative to sulfate), and (b) reductions in the leachate acidity and ammonium relative to the incident droplets. A relationship was observed between the enhancement of Na, Ca, and nitrate in pine needle leachate. This suggests that reactions at the foliar surfaces are occurring which involve gaseous HNO_3 and accumulated soil dust and sea-salt particles.