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W Haase

Publications and source records attributed to W Haase.

At least 127 records · Page 7Linked to original sources
PubMed1988

Does sphingomyelin inhibit the erythrocyte anion transport system?

The anion transport protein of the human erythrocyte membrane, band 3, was incorporated into unilamellar sphingomyelin vesicles. The vesicles showed a rapid sulfate efflux which could be inhibited by specific inhibitors of the erythrocyte anion transport system. All band 3 molecules contributing to the inhibitor-sensitive flux component were arranged 'right-side-out'. The turnover number of the transport protein for sulfate transport was virtually identical to that in phosphatidylcholine bilayers and around 6 times larger than in human erythrocyte membranes. Thus, in contrast to other claims, sphingomyelin does not inhibit the erythrocyte anion transport system.

Anion Exchange Protein 1, Erythrocyte↗
PubMed1988

The turnover number for band 3-mediated sulfate transport in phosphatidylcholine bilayers.

The anion transport system of the human erythrocyte membrane was reconstituted in unilamellar phosphatidylcholine vesicles, and a vesicle subpopulation of a narrow size distribution was isolated from the sample by gel filtration. In this subpopulation, the turnover number of the transport protein (the band 3 protein) for sulfate transport was determined. It was found that, in the reconstituted system, the protein transports sulfate 5-10 times faster than in the human erythrocyte membrane.

Anion Exchange Protein 1, Erythrocyte↗
PubMed1987

[Mammography after breast-preserving therapy of breast cancer].

Follow-up after tumorectomy and intensive irradiation for carcinoma of the breast creates new diagnostic problems. In particular, more experience is necessary about the value of mammography. We have therefore analysed the mammograms of 104 patients intensively followed over one to eight years after primary treatment. Changes in the appearances of the breast due to irradiation are compared with mammographic changes caused by recurrence of tumor. It can be shown that definite structural patterns can be recognised regularly. Optimal diagnosis of recurrencies, however, is only possible by comparing all follow-up mammograms, and additionally the pre-operative films. Close cooperation between the surgeon, radiotherapist and diagnostic radiologist is essential.

Breast Neoplasms↗
PubMed1987

[Diplopia, enophthalmos and motility disorders in isolated orbital floor fractures].

In cases of pure blow-out fracture with unequivocal clinical symptoms (limitation of movement of the globe, dislocation and/or enophthalmos) and confirmed X-ray findings, early surgery is the treatment of choice. In cases where the clinical findings are less pronounced, a "wait-and-see" policy, keeping the patient under observation, seems appropriate for the sake of avoiding surgical trauma. Coronal computerized tomography offers advantages in cases where the diagnosis is questionable.

Adult↗
PubMed1987

Reconstitution of the erythrocyte anion transport system: recent progress.

The anion transport system of the human erythrocyte membrane was reconstituted in phosphatidylcholine vesicles by a new procedure. The reconstituted system shows all major properties of the sulfate transport system found in the erythrocyte membrane (Scheuring, U., K. Kollewe, W. Haase, and D. Schubert: J. Membrane Biol. 90, 123-135 (1986)). Modifications of the reconstitution method have now led to increased values of the internal volumes of the protein-lipid vesicles and of the average turnover number of the transport protein (which is now larger than in the erythrocyte membrane). Gel filtration on Sephacryl S-1000 has allowed the isolation of vesicles of a narrow size distribution.

Anions↗
PubMed1986

A new method for the reconstitution of the anion transport system of the human erythrocyte membrane.

The anion transport protein of the human erythrocyte membrane, band 3, was solubilized and purified in solutions of the non-ionic detergent Triton X-100. It was incorporated into spherical lipid bilayers by the following procedure: Dry phosphatidylcholine was suspended in the protein solution. Octylglucopyranoside was added until the milky suspension became clear. The sample was dialyzed overnight against detergent-free buffer. Residual Triton X-100 was removed from the opalescent vesicle suspension by sucrose density gradient centrifugation and subsequent dialysis. Sulfate efflux from the vesicles was studied, under exchange conditions, using a filtration method. Three vesicle subpopulations could be distinguished by analyzing the time course of the efflux. One was nearly impermeable to sulfate, and efflux from another was due to leaks. The largest subpopulation, however, showed transport characteristics very similar to those of the anion transport system of the intact erythrocyte membrane: transport numbers (at 30 degrees C) close to 20 sulfate molecules per band 3 and min, an activation energy of approx. 140 kJ/mol, a pH maximum at pH 6.2, saturation of the sulfate flux at sulfate concentrations around 100 mM, inhibition of the flux by H2DIDS and flufenamate (approx. KI-values at 30 degrees C: 0.1 and 0.7 microM, respectively), and "right-side-out" orientation of the transport protein (as judged from the inhibition of sulfate efflux by up to 98% by externally added H2DIDS). Thus, the system represents, for the first time, a reconstitution of all the major properties of the sulfate transport across the erythrocyte membrane.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗
PubMed1985

Na+/Ca2+ countertransport in plasma membrane of rat pancreatic acinar cells.

The presence of a coupled Na+/Ca2+ exchange system has been demonstrated in plasma membrane vesicles from rat pancreatic acinar cells. Na+/Ca2+ exchange was investigated by measuring 45Ca2+ uptake and 45Ca2+ efflux in the presence of sodium gradients and at different electrical potential differences across the membrane (= delta phi) in the presence of sodium. Plasma membranes were prepared by a MgCl2 precipitation method and characterized by marker enzyme distribution. When compared to the total homogenate, the typical marker for the plasma membrane, (Na+ + K+)-ATPase was enriched by 23-fold. Markers for the endoplasmic reticulum, such as RNA and NADPH cytochrome c reductase, as well as for mitochondria, the cytochrome c oxidase, were reduced by twofold, threefold and 10-fold, respectively. For the Na+/Ca2+ countertransport system, the Ca2+ uptake after 1 min of incubation was half-maximal at 0.62 mumol/liter Ca2+ and at 20 mmol/liter Na+ concentration and maximal at 10 mumol/liter Ca2+ and 150 mmol/liter Na+ concentration, respectively. When Na+ was replaced by Li+, maximal Ca2+ uptake was 75% as compared to that in the presence of Na+. Amiloride (10(-3) mol/liter) at 200 mmol/liter Na+ did not inhibit Na+/Ca2+ countertransport, whereas at low Na+ concentration (25 mmol/liter) amiloride exhibited dose-dependent inhibition to be 62% at 10(-2) mol/liter. CFCCP (10(-5) mol/liter) did not influence Na+/Ca2+ countertransport. Monensin inhibited dose dependently; at a concentration of 5 X 10(-6) mol/liter inhibition was 80%. A SCN- or K+ diffusion potential (= delta phi), being positive at the vesicle inside, stimulated calcium uptake in the presence of sodium suggesting that Na+/Ca2+ countertransport operates electrogenically, i.e. with a stoichiometry higher than 2 Na+ for 1 Ca2+. In the absence of Na+, delta phi did not promote Ca2+ uptake. We conclude that in addition to ATP-dependent Ca2+ outward transport as characterized previously (E. Bayerdörffer, L. Eckhardt, W. Haase & I. Schulz, 1985, J. Membrane Biol. 84:45-60) the Na+/Ca2+ countertransport system, as characterized in this study, represents a second transport system for the extrusion of calcium from the cell. Furthermore, the high affinity for calcium suggests that this system might participate in the regulation of the cytosolic free Ca2+ level.

Adenosine Triphosphatases↗
PubMed1985

Electrogenic calcium transport in plasma membrane of rat pancreatic acinar cells.

ATP-dependent 45Ca2+ uptake was investigated in purified plasma membranes from rat pancreatic acinar cells. Plasma membranes were purified by four subsequent precipitations with MgCl2 and characterized by marker enzyme distribution. When compared to the total homogenate, typical marker enzymes for the plasma membrane, (Na+,K+)-ATPase, basal adenylate cyclase and CCK-OP-stimulated adenylate cyclase were enriched by 43-fold, 44-fold, and 45-fold, respectively. The marker for the rough endoplasmic reticulum was decreased by fourfold compared to the total homogenate. Comparing plasma membranes with rough endoplasmic reticulum, Ca2+ uptake was maximal with 10 and 2 mumol/liter free Ca2+, and half-maximal with 0.9 and 0.5 mumol/liter free Ca2+. It was maximal at 3 and 0.2 mmol/liter free Mg2+ concentration, at an ATP concentration of 5 and 1 mmol/liter, respectively, and at pH 7 for both preparations. When Mg2+ was replaced by Mn2+ or Zn2+ ATP-dependent Ca2+ uptake was 63 and 11%, respectively, in plasma membranes; in rough endoplasmic reticulum only Mn2+ could replace Mg2+ for Ca2+ uptake by 20%. Other divalent cations such as Ba2+ and Sr2+ could not replace Mg2+ in Ca2+ uptake. Ca2+ uptake into plasma membranes was not enhanced by oxalate in contrast to Ca2+ uptake in rough endoplasmic reticulum which was stimulated by 7.3-fold. Both plasma membranes and rough endoplasmic reticulum showed cation and anion dependencies of Ca2+ uptake. The sequence was K+ greater than Rb+ greater than Na+ greater than Li+ greater than choline+ in plasma membranes and Rb+ greater than or equal to K+ greater than or equal to Na+ greater than Li+ greater than choline+ for rough endoplasmic reticulum. The anion sequence was Cl greater than or equal to Br greater than or equal to 1 greater than SCN greater than NO3 greater than isethionate greater than cyclamate greater than gluconate greater than SO2(4) greater than or equal to glutarate and Cl- greater than Br greater than gluconate greater than SO2(4) greater than NO3 greater than 1 greater than cyclamate greater than or equal to SCN, respectively. Ca2+ uptake into plasma membranes appeared to be electrogenic since it was stimulated by an inside-negative K+ and SCN diffusion potential and inhibited by an inside-positive diffusion potential. Ca2+ uptake into rough endoplasmic reticulum was not affected by diffusion potentials. We assume that the Ca2+ transport mechanism in plasma membranes as characterized in this study represents the extrusion system for Ca2+ from the cell that might be involved in the regulation of the cytosolic Ca2+ level.

Adenosine Triphosphate↗
PubMed1985

[Postoperative microstrabismus: performance accuracy in spatial depth].

Thirty-six patients with postoperative microstrabismus (24 of them without or with only a slight amblyopia, 12 with severe unilateral amblyopia) and three control groups (normal binocular, long-term monocular, and inexperienced monocular subjects) were tested by their ability to hit 14 vertical sticks set in varied positions in a rectangular board, without any time limit during the first test and with time limits in the second and third tests. The evaluation shows that all 36 patients achieved considerably better results than any of the monocular subjects, but did not score as highly as normal binocular persons.

Adolescent↗
PubMed1985

ATP-dependent H+ pump in membrane vesicles from rat kidney cortex.

The presence of membrane vesicles containing an ATP-driven H+ pump was demonstrated in rat kidney cortex homogenate using the delta pH-sensitive dye acridine orange (AO). These vesicles were purified by differential and Percoll density gradient centrifugation. ATP-driven H+ uptake was about 20-fold enriched compared with the homogenate. Determination of marker enzyme activities indicated that these vesicles do not originate from brush border and basolateral membranes, lysosomes, endoplasmic reticulum, mitochondria, Golgi membranes, or red blood cells. The identity with brush border membranes was further excluded by the absence of Na+-H+ exchange. Renal cortical endocytotic vesicles that had taken up horseradish peroxidase or fluorescein isothiocyanate-labeled dextran (FITC-dextran) after injection of these substances into rats in vivo comigrated with the H+ pump activity on the Percoll gradient. Similar characteristics of the H+ pump demonstrated by the AO method and by fluorescence changes of in vivo trapped FITC-dextran proved the identity of H+ pump-containing vesicles with endocytotic vesicles. ATP-driven H+ uptake into endocytotic vesicles was stimulated by Cl- and weakly inhibited by oligomycin. N-ethylmaleimide, dicyclohexylcarbodiimide, and Dio-9 were stronger inhibitors. Histochemical studies revealed that horseradish peroxidase-filled endocytotic vesicles are localized in the apical region of proximal tubule cells. An H+ pump with similar characteristics, but much lower activity, was found in brush border membranes, basolateral membranes, and mitochondria isolated by standard techniques, suggesting a possible contamination of these preparations with endocytotic vesicles.

Acridine Orange↗
PubMed1984

Reconstitution and partial purification of several Na+ cotransport systems from renal brush-border membranes. Properties of the L-glutamate transporter in proteoliposomes.

Brush-border membranes of renal proximal tubules were solubilized with deoxycholate and some proteins were separated and incorporated into proteoliposomes by a reconstitution procedure which was analyzed in detail. The proteoliposomes contained mainly polypeptides with molecular weights of 152,000, 94,000, and 52,000, each of which could be separated further into homologous polypeptides with different isoelectric points. In the proteoliposomes, Na+ cotransport systems for D-glucose, acidic and neutral amino acids, and mono- and dicarboxylic acids were demonstrated by showing that due to an inwardly directed Na+ gradient the substrate concentrations in the proteoliposomes increased significantly over their respective equilibrium values. Using inhibition experiments, selectivity of the different transporters could be demonstrated. Studying the reconstituted L-glutamate transporter in detail, countertransport of L-glutamate and K+ was shown (i) at Na+ equilibrium the intraliposomal L-glutamate concentration increased significantly over the equilibrium value if an outside-directed K+ gradient was applied; (ii) Rb+ influx was significantly stimulated by the outflux of L-glutamate. By applying a K+ diffusion potential across the liposomal membrane by addition of valinomycin it could be shown that during L-glutamate transport in the presence of Na+ and K+ positive charge is transferred together with L-glutamate and Na+. The apparent Km value of L-glutamate uptake driven by concentration differences of 89 mM Na+ (out greater than in) and 89 mM K+ (in greater than out) was 26.3 +/- 1.3 microM. The Vmax value of 70.2 +/- 2.3 pmol X mg of protein-1 X S-1 was half the value measured in intact membranes.

Amino Acid Transport System X-AG↗
PubMed1984

[Treatment of coronary disease with elantan. Results of a multicenter study].

10,229 patients with coronary heart disease (CHD) took part in a multicenter study to verify the efficacy and tolerance of a 14 days lasting therapy with elantan under practical conditions. The majority of patients with an average age of 59 +/- 10 years (58.8% men, 40.3% women) had suffered about four years on CHD and had already been treated with coronary efficient drugs. The 14-day therapy with elantan 20 in a dosage of 3 X 1 tabl./day showed a cure rate of 79.9% in patients who had already been treated, a complete remission of the angina pectoris attacks being achieved in 50% and a reduction of the frequency of the attacks in a further 29.9%. The reduced frequency of the attacks was associated with a reduced consumption of acute nitrates (nitroglycerine capsules). With the smaller group of patients who were being treated for angina pectoris for the first time, the therapy with elantan was characterized by a cure rate of 91%.

Angina Pectoris↗
PubMed1984

Electron-microscopic demonstration of the distribution of calcium deposits in the exocrine pancreas of the rat after application of carbachol, atropine, cholecystokinin, and procaine.

In an attempt to identify a cellular Ca2+-pool, from which calcium is released when secretagogues are applied, tissue fragments of the rat exocrine pancreas were incubated and fixed with glutaraldehyde in the presence of calcium. By means of this procedure electron-dense deposits were found on plasma membranes. X-ray microanalysis showed that these deposits contain calcium. Stimulation of tissue fragments with the use of the secretagogues carbachol or cholecystokinin reduced the number of deposits by about 80%. When the antagonist atropine was applied after carbachol stimulation, deposits reappeared on cell membranes, which then disappeared again after a second stimulation with cholecystokinin. In the presence of procaine, carbachol was inhibited and only slightly reduced the Ca2+-deposits on the plasma membranes. These results suggest that a calcium pool, from which calcium is released to induce enzyme secretion on stimulation, is located in the cell membrane.

Animals↗
PubMed1984

A stopped flow capillary perfusion method to evaluate contraluminal transport parameters of methylsuccinate from interstitium into renal proximal tubular cells.

In order to study the transport of dicarboxylic acids through the contraluminal cell membrane of proximal tubular cells, 3H- methylsuccinate has been synthetized by catalytic hydration of methylfumarate . As the chromatography of radioactive material excreted in the urine after i.v. injection of 3H- methylsuccinate shows, no metabolite is detectable during the first 3 min. After 10 min, less than 10% of the excreted radiolabel is metabolized. To measure the contraluminal influx of 3H- methylsuccinate from the interstitium into cortical tubular cells, the renal vessels were clamped so that the proximal tubular lumina collapsed. Then Ringer solution was injected into the blood capillaries. It contained different concentrations of 3H- methylsuccinate and 14C-inulin as extracellular space marker. After contact times between 1 and 10 s, this fluid was withdrawn from the capillaries and the disappearance of 3H- methylsuccinate relative to 14C-inulin was measured. The morphological compartments in the outer cortex of the clamped glutaraldehyde-fixed kidney were evaluated by a stereological method. For proximal tubular cells a ratio of extracellular water space to intracellular space of 1:3.1 and a ratio extracellular water space to free cell water space of 1:2 was found. It was tested whether the experimental disappearance curves with 4 different starting concentrations of 3H- methylsuccinate fit with the data from four model calculations. It was found that the data and the conditions of transport are consistent with the predictions of a facilitated diffusion model.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

Effect of inositol-1,4,5-trisphosphate on isolated subcellular fractions of rat pancreas.

We have previously shown that inositol-1,4,5-trisphosphate (IP3) releases Ca2+ from an intracellular calcium store in permeabilized acinar cells of rat pancreas (H. Streb et al., 1983, Nature (London) 306:67-69). This observation suggests that IP3 might provide the missing link between activation of the muscarinic receptor and Ca2+ release from intracellular stores during stimulation. In order to localize the intracellular IP3-sensitive calcium pool, IP3-induced Ca2+ release was measured in isolated subcellular fractions. A total homogenate was prepared from acinar cells which had been isolated by a collagenase digestion method. Endoplasmic reticulum was separated from mitochondria, zymogen granules and nuclei by differential centrifugation. Plasma membranes and endoplasmic reticulum were separated by centrifugation on a sucrose step gradient or by precipitation with high concentrations of MgCl2. IP3-induced Ca2+ release per mg protein in the total homogenate was the same as in leaky cells and was sufficiently stable to make short separation procedures possible. In fractions obtained by either differential centrifugation at 7000 X g, sucrose-density centrifugation, or MgCl2 precipitation there was a close correlation of Ip3-induced Ca2+ release with the endoplasmic reticulum markers ribonucleic acid (r = 0.96, 1.00, 0.91, respectively) and NADPH cytochrome c reductase (r = 0.63, 0.98, 0.90, respectively). In contrast, there was a clear negative correlation with the mitochondrial markers cytochrome c oxidase (r = -0.64) and glutamate dehydrogenase (r = -0.75) and with the plasma membrane markers (Na+ + K+)-ATPase (r = -0.81) and alkaline phosphatase (r = -0.77) in all fractions analyzed. IP3-induced Ca2+ release was distributed independently of zymogen granule or nuclei content of the fractions as assessed by electron microscopy. The data suggest that inositol-1,4,5-trisphosphate releases Ca2+ from endoplasmic reticulum in pancreatic acinar cells.

Animals↗