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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 102 (1988), S. 247-254 
    ISSN: 1432-1424
    Keywords: exocrine gland ; parotid ; acinar cell ; ion transport ; fluid secretion ; pH regulation ; antiport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary pH gradient-dependent sodium transport in highly purified rat parotid basolateral membrane vesicles was studied under voltage-clamped conditions. In the presence of an outwardly directed H+ gradient (pHin=6.0, pHout=8.0)22Na uptake was approximately ten times greater than uptake measured at pH equilibrium (pHin=pHout=6.0). More than 90% of this sodium flux was inhibited by the potassium-sparing diuretic drug amiloride (K 1 =1.6 μm) while the transport inhibitors furosemide (1mm), bumetanide (1mm) SITS (0.5mm) and DIDS (0.1mm) were without effect. This transport activity copurified with the basolateral membrane marker K+-stimulatedp-nitrophenyl phosphatase. In addition22Na uptake into the vesicles could be driven against a concentration gradient by an outwardly directed H+ gradient. pH gradient-dependent sodium flux exhibited a simple Michaelis-Menten-type dependence on sodium concentration cosistent with the existence of a single transport system withK M =8.0mm at 23°C. A component of pH gradient-dependent, amiloride-sensitive sodium flux was also observed in rabbit parotid basolateral membrane vesicles. These results provide strong evidence for the existence of a Na+/H+ antiport in rat and rabbit parotid acinar basolateral membranes and extend earlier less direct studies which suggested that such a transporter was present in salivary acinar cells and might play a significant role in salivary fluid secretion.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 120 (1991), S. 125-130 
    ISSN: 1432-1424
    Keywords: loop diuretics ; exocrine gland ; fluid secretion ; lipid ; acinar cell ; ion transport ; chloride secretion ; detergent
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary It was recently reported (Turner, R.J., George, J.N., 1990,J. Membrane Biol. 113:203–210) that the high affinity bumetanide binding site of the rabbit parotid Na/K/Cl cotransporter could be extracted from a basolateral membrane preparation from this gland using relatively low concentrations of the non-ionic detergent Triton X-100. At the detergent: protein ratios required for complete membrane solubilization bumetanide binding activity in this extract was lost but could be recovered by the addition of crude soybean lipids. In the present paper the ability of various purified lipids to restore high affinity bumetanide binding activity in detergent solubilized rabbit parotid basolateral membranes is studied. We show that the effect of exogenous lipid on the detergent-inactivated bumetanide binding site is to increase the affinity of binding without affecting the number of binding sites. Of the 11 lipid species tested, several relatively minor, negatively charged membrane phospholipids are the most effective in restoring binding activity (phosphatidylserine ≈ phosphatidylglycerol 〉 phosphatidylinositol 〉 cardiolipin). while the major mammalian plasma membrane lipid components phosphatidylcholine, phosphatidylethanolamine, sphingomyelin and cholesterol are without effect. In addition, we show that in the presence of these minor lipids the affinity of bumetanide binding is considerably increased over that observed in the native membrane (e.g.,K d ≈0.06 μm in membranes extracted with 0.3% Triton and treated with 0.15% wt/vol phosphatidylserine,vs. K d ≈3 μm in native basolateral membranes). This dramatic dependence of bumetanide binding affinity on the presence of certain lipid species suggests that the properties of the bumetanide binding proteinin situ may be quite dependent on the minor lipid content of the plasma membrane. This effect may account for the relatively large variations in bumetanide binding affinity observed from tissue to tissue.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 102 (1988), S. 71-77 
    ISSN: 1432-1424
    Keywords: loop diuretics ; exocrine gland ; fluid secretion ; parotid ; acinar cell ; ion transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The Na/K/Cl-dependent component of the binding of the loop diuretic bumetanide to basolateral membrane vesicles from the rabbit parotid is studied. A Scatchard analysis indicates that this binding is due to a single high-affinity site withK D =3.2±0.3 μm (n=9) at 100mm sodium, 100mm potassium and 5mm chloride. When KCl-dependent22Na transport and tracer [3H]-bumetanide binding are monitored simultaneously as a function of (unlabeled) bumetanide concentration it is found that theK 0.5 for bumetanide inhibition of both processes are identical indicating that the high-affinity bumetanide binding site studied here is identical with a bumetanide-inhibitory site on the Na/K/Cl cotransport system previously identified in this preparation (R.J. Turner, J.N. George and B.J. Baum,J. Membrane Biol. 94:143–152, 1986). High-affinity bumetanide binding exhibits a hyperbolic dependence on both [Na] and [K] consistent with Na/bumetanide and K/bumetanide binding stoichiometries of 1∶1 andK 0.5 values of approximately 33mm for sodium and 23mm for potassium. In contrast, the dependence on [Cl] is biphasic, with bumetanide binding increasing from 0 to 5mm chloride and decreasing toward baseline levels thereafter. Scatchard analysis of this latter inhibitory effect of chloride indicates a competitive interaction with bumetanide in agreement with earlier indications that bumetanide inhibits Na/K/Cl cotransport at a chloride site. However, studies of the effects of various anions on bumetanide binding and22Na transport show a poor correlation between the specificities of these two processes, suggesting that the inhibitory chloride site is not a chloride transport site.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 111 (1989), S. 191-198 
    ISSN: 1432-1424
    Keywords: exocrine gland ; pH regulation ; Na+/H+ exchange ; fluid secretion ; muscarinic agonist
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The present studies were designed to test our previous suggestion that Na+/H+ exchange was activated by muscarinic stimulation of rat parotid acinar cells. Consistent with this hypothesis, we demonstrate here that intact rat parotid acini stimulated with the muscarinic agonist carbachol in HCO 3 − -free medium show an enhanced recovery from an acute acid load as compared to similarly challenged untreated preparations. Amiloride-sensitive22Na uptake, due to Na+/H+ exchange, was also studied in plasma membrane vesicles prepared from rat parotid acini pretreated with carbachol. This uptake was stimulated twofold relative to that observed in vesicles from control (untreated) acini. This stimulation was time dependent, requiring ∼15 min of acinar incubation with carbachol to reach completion, and ws blocked by the presence of the muscarinic antagonist atropine (2×10−5 m) in the pretreatment medium. The effect of carbachol was dose dependent withK 0.5∼3×10−6 m. Stimulation of the exchanger was also seen in vesicles prepared from acini pretreated with the α-adrenergic agonist epinephrine, but not with the β-adrenergic agonist isoproterenol, or with substance P. Kinetic analysis indicated that the stimulation induced by carbachol was due to an alkaline shift in the pH responsiveness of the exchanger in addition to an increasedapparent transport capacity. Taken together with previous results from this and other laboratories, these results strongly suggest that the Na+/H+ exchanger and its regulation are intimately involved in the fluidsecretory response of the rat parotid.
    Type of Medium: Electronic Resource
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 112 (1989), S. 51-58 
    ISSN: 1432-1424
    Keywords: loop diuretics ; exocrine gland ; fluid secretion ; parotid ; acinar cell ; ion transport ; chloride secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary The inactivation of the rabbit parotid Na/K/Cl cotransporter by the irreversible sulfhydryl reagent N-ethylmaleimide (NEM) is studied by monitoring its effect on high affinity bumetanide binding to the carrier. NEM reduces the number of bumetanide binding sites with no significant change in the affinity of those remaining. NEM also reduces KCl-dependent22Na flux via the cotransporter by the same factor as the reduction in bumetanide binding sites. Both bumetanide and its analogue furosemide can protect against the effect of NEM. The concentration range over which this protection occurs is in good agreement with affinities of these two compounds for the high affinity bumetanide binding site (2.6 and 85 μm, respectively), indicating an association of this site with the site of action of NEM. Also consistent with this hypothesis are the observations that (i) sodium and potassium, both of which are required for high affinity bumetanide binding, increase the rate of inactivation of binding by NEM and (ii) chloride, at concentrations previously shown to competitively inhibit bumetanide binding, protects the cotransporter against NEM. The effects of NEM on bumetanide binding are mimicked by another highly specific sulfhydryl reagent, methyl methanethiolsulfonate. The apparent rate constant for inactivation of high affinity bumetanide binding by NEM is a hyperbolic function of NEM concentration consistent with a model in which the inactivation reaction is first order in [NEM] and proceeds through an intermediate adsorptive complex. The data indicate that the presence of a reduced sulfhydryl group at or closely related to the bumetanide binding site is essential for the operation of the parotid Na/K/Cl cotransporter.
    Type of Medium: Electronic Resource
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  • 6
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 113 (1990), S. 203-210 
    ISSN: 1432-1424
    Keywords: loop diuretics ; exocrine gland ; fluid secretion ; parotid ; acinar cell ; ion transport ; chloride secretion ; detergent
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary We demonstrate that the high affinity bumetanide binding site of the rabbit parotid acinar cell can be extracted from a basolateral membrane fraction using relatively low concentrations (0.07%, wt/vol; 1 mg membrane protein/ml) of the nonionic detergent Triton X-100. This extracted site cannot be sedimented by ultracentrifugation at 100,000 ×g × 1 hr. Bumetanide binding to this site retains the ionic characteristics of bumetanide binding to native membranes but shows a fivefold increase in binding affinity (K d=0.57±0.15 μm vs.K d=3.3±0.7 μm for native membranes). Inactivation of the extracted bumetanide binding site observed at detergent/protein ratios〉1 can be prevented or (partially) reversed by the addition of exogenous lipid (0.2% soybean phosphatidylcholine). When the 0.07% Triton extract is fractionated by sucrose density gradient centrifugation in 0.24% Triton X-100, 0.2% exogenous lipid and 200mm salt, the high affinity bumetanide binding site sediments as a single band withS 20,w =8.8±0.8 S. This corresponds to a molecular weight ∼200 kDa for the bumetanide binding protein-detergent-lipid complex and represents a sevenfold purification of this site relative to the starting membrane fraction. In contrast to previous attempts to purify Na/K/Cl cotransport proteins and their associated bumetanide binding sites, the present method avoids harsh detergent treatment as well as direct covalent modification (inactivation) of the transporter itself. As a consequence, one can follow the still active protein through a series of extraction and purification steps by directly monitoring its bumetanide binding properties.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    The journal of membrane biology 94 (1986), S. 143-152 
    ISSN: 1432-1424
    Keywords: fluid secretion ; exocrine gland ; chloride transport
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Summary Sodium (22Na) transport was studied in a basolateral membrane vesicle preparation from rabbit parotid. Sodium uptake was markedly dependent on the presence of both K+ and Cl− in the extravesicular medium, being reduced 5 times when K+ was replaced by a nonphysiologic cation and 10 times when Cl− was replaced by a nonphysiologic anion. Sodium uptake was stimulated by gradients of either K+ or Cl− (relative to nongradient conditions) and could be driven against a sodium concentration gradient by a KCl gradient. No effect of membrane potentials on KCl-dependent sodium flux could be detected, indicating that this is an electroneutral process. A KCl-dependent component of sodium flux could also be demonstrated under equuilibrium exchange conditions, indicating a direct effect of K+ and Cl− on the sodium transport pathway. KCl-dependent sodium uptake exhibited a hyperbolic dependence on sodium concentration consistent with the existence of a single-transport system withK m =3.2mm at 80mm KCl and 23°C. Furosemide inhibited this transporter withK 0.5=2×10−4 m (23°C). When sodium uptake was measured as a function of potassium and chloride concentrations a hyperbolic dependence on [K] (Hill coefficient =1.31±0.07) were observed, consistent with a Na/K/Cl stoichiometry of 1∶1∶2. Taken together these data provide strong evidence for the electroneutral coupling of sodium and KCl movements in this preparation and strongly support the hypothesis that a Na+/K+/Cl− cotransport system thought to be associated with transepithelial chloride and water movements in many exocrine glands is present in the parotid acinar basolateral membrane.
    Type of Medium: Electronic Resource
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Environmental management 7 (1983), S. 433-442 
    ISSN: 1432-1009
    Keywords: Coastal zone management ; Wetlands ; Canals ; Marshes ; Louisiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Annual coastal land loss in the sedimentary deltaic plain of southern Louisiana is 102 km2, which is correlated with man-made canal surface area. The relationships between land loss and canals are both direct and indirect and are modified by the deltaic substrate, distance to the coast, and availability of new sediments. Loss rates are highest in the youngest of the former deltas nearest the coast; they are lowest in the more consolidated sediments far from the coast. The average estimate for land loss at zero canal density in the six regression equations developed was 0.09%±0.13% annually, the present land loss rates approach 0 8% annually Although additional analyses are needed, we conclude that canals are causally related to a significant portion of the total coastal land loss rates The relation probably involves an interruption of local and regional hydrologic regimes. Reduction of the present acceleration in land loss rates is possible by managing present canals more effectively, by not permitting new ones, and by changing the design of new canals to allow more natural water flow
    Type of Medium: Electronic Resource
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  • 9
    Electronic Resource
    Electronic Resource
    Springer
    Environmental management 11 (1987), S. 823-836 
    ISSN: 1432-1009
    Keywords: Backfilling ; Mitigation ; Wetlands ; Louisiana ; Dredging
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract Returning canal spoil banks into canals, or backfilling, is used in Louisiana marshes to mitigate damage caused by dredging for oil and gas extraction. We evaluated 33 canals backfilled through July 1984 to assess the success of habitat restoration. We determined restoration success by examining canal depth, vegetation recolonization, and regraded spoil bank soils after backfilling. Restoration success depended on: marsh type, canal location, canal age, marsh soil characteristics, the presence or absence of a plug at the canal mouth, whether mitigation was on- or off-site, and dredge operator performance. Backfilling reduced median canal depth from 2.4 to 1.1 m, restored marsh vegetation on the backfilled spoil bank, but did not restore emergent marsh vegetation in the canal because of the lack of sufficient spoil material to fill the canal and time. Median percentage of cover of marsh vegetation on the canal spoil banks was 51.6%. Median percentage of cover in the canal was 0.7%. The organic matter and water content of spoil bank soils were restored to values intermediate between spoil bank levels and predredging marsh conditions. The average percentage of cover of marsh vegetation on backfilled spoil banks was highest in intermediate marshes (68.6%) and lowest in fresh (34.7%) and salt marshes (33.9%). Average canal depth was greatest in intermediate marshes (1.50 m) and least in fresh marshes (0.85 m). Canals backfilled in the Chenier Plain of western Louisiana were shallower (average depth = 0.61 m) than in the eastern Deltaic Plain (mean depth range = 1.08 to 1.30 m), probably because of differences in sediment type, lower subsidence rate, and lower tidal exchange in the Chenier Plain. Canals backfilled in marshes with more organic soils were deeper, probably as a result of greater loss of spoil volume caused by oxidation of soil organic matter. Canals ten or more years old at the time of backfilling had shallower depths after backfilling. Depths varied widely among canals backfilled within ten years of dredging. Canal size showed no relationship to canal depth or amount of vegetation reestablished. Plugged canals contained more marsh reestablished in the canal and much greater chance of colonization by submerged aquatic vegetation compared with unplugged canals. Dredge operator skill was important in leveling spoil banks to allow vegetation reestablishment. Wide variation in dredge performance led to differing success of vegetation restoration. Complete reestablishment of the vegetation was not a necessary condition for successful restoration. In addition to providing vegetation reestablishment, backfilling canals resulted in shallow water areas with higher habitat value for benthos, fish, and waterfowl than unfilled canals. Spoil bank removal also may help restore water flow patterns over the marsh surface. Increased backfilling for wetland mitigation and restoration is recommended.
    Type of Medium: Electronic Resource
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Environmental management 18 (1994), S. 271-282 
    ISSN: 1432-1009
    Keywords: Wetland ; Restoration ; Dredging ; Spoil ; Louisiana
    Source: Springer Online Journal Archives 1860-2000
    Topics: Energy, Environment Protection, Nuclear Power Engineering
    Notes: Abstract The rationale and outline of an implementation plan for restoring coastal wetlands in Louisiana is presented. The rationale for the plan is based on reversing the consequences of documented cause-and-effect relationships between wetland loss and hydrologic change. The main feature is to modify the extensive interlocking network of dredged spoil deposits, or spoil banks, by reestablishing a more natural water flow at moderate flow velocity (〈5 cm/sec). Guidelines for site selection from thousands of potential sites are proposed. Examples of suitable sites are given for intermediate marshes. These sites exhibit rapid deterioration following partial or complete hydrologic impoundment, implying a strong hydrologic, rather than sedimentological, cause of wetland deterioration. We used an exploratory hydrologic model to guide determination of the amount of spoil bank to be removed. The results from an economic model indicated a very effective cost-benefit ratio. Both models and practical experience with other types of restoration plans, in Louisiana and elsewhere, exhibit an economy of scale, wherein larger projects are more cost effective than smaller projects. However, in contrast to these other projects, spoil bank management may be 100 to 1000 times more cost effective and useful in wetland tracts 〈1000 ha in size. Modest spoil bank management at numerous small wetland sites appears to offer substantial positive attributes compared to alternative and more intensive management at a few larger wetland sites.
    Type of Medium: Electronic Resource
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