Universität Stuttgart
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Item Open Access Extraordinary biological membrane structures resulting from different local membrane curvatures(1992) Meyer, Helmut W.; Hülser, Dieter F.The bilayer arrangement of amphiphilic molecules is not only the basic structure of rather flat biological membranes, but also of regularly curved bilayers in most cubic phase structures. The basis of these cubic phase structures are infinite periodical minimal surfaces (IPMS). Extraordinary biological membrane structures resembling such IPMS were found as periodically curved bilayers in areas of the plasma membrane in a Streptomyces strain and in liposomes prepared from its extracted lipids. This structure consists of a transition of convex to concave curvatures and vice versa. A structure with curvatures in one direction only was observed in vacuolar membranes of yeast cells with a genetic defect. Our electron microscopical analysis of freeze fractured membranes of these cells revealed not only fully invaginated but also flat particle-free areas which were mainly circularly shaped, some elongated areas, however, were also present. In addition, sometimes periodical arrangements were detected which obviously are not related to IPMS structures. Both structures, however, indicate a high proportion of wedge-shaped lipid molecules in the bilayer.Item Open Access The use of the tip potential of glass microelectrodes in the determination of low cell membrane potentials(1973) Hülser, Dieter F.; Webb, Dennis J.The tip potential of Ling-Gerard glass microelectrodes changes upon insertion into cells and thus impedes the determination of the actual membrane potential. The lower the membrane potential of a cell, the larger will be the error due to this tip potential. However, as is demonstrated, a relationship exists between the tip potential of the electrode and the measured potential difference, which allows the determination of the membrane potential of a particular cell type by linear regression. This method showed that resting lymphocytes had no membrane potential, whereas for the slime mould Dictyostelitium discoideum a membrane potential of about -9 mV could be calculated.Item Open Access Microbial metabolism of chlorosalicylates: effect of prolonged subcultivation on constructed strains(1986) Rubio, Miguel Angel; Engesser, Karl-Heinrich; Knackmuss, Hans-JoachimThe hybrid strain Pseudomonas sp. WR4016 was subcultivated with increasing concentrations of 5-chlorosalicylate (5rarr10 mM) as sole carbon source over a period of 9 months. At intervals of approximately 3 months derivative strains WR4017, WR4018 and WR4019 were isolated which exhibited higher growth rates and increased substrate tolerance. Comparative analysis of the turnover rates of the key enzymes in chlorosalicylate degradation showed that the adaptation process did not result from structural modifications of these proteins. Instead, balanced over-production of the salicylate hydroxylase and catechol 1,2-dioxygenase prevented the accumulation of toxic chlorocatechols and accounted for the reduction of the doubling times with 4- or 5-chlorosalicylate. A comparative analysis of a genetically engineered chlorosalicylate degrader PL300-1 showed similar regulatory patterns as the most advanced isolate WR4019 from the adaptation series.Item Open Access Reduced cavitation-induced cellular damage by the antioxidative effect of vitamin E(1994) Suhr, Dierk; Brümmer, Franz; Irmer, Ulrich; Schlachter, Manfred; Hülser, Dieter F.Fragmentation of human urinary and biliary stones by shock waves in extracorporeal lithotripsy is accompanied by tissue damage. Both the fragmentation as well as the side effects are often attributed to cavitation. The hazardous potential of cavitation is not only of a physical nature but also of a chemical nature, because of the generation of free radicals, e.g. ·OH, ·H and ·O2. After the application of shock waves, we have demonstrated cavitation-generated free radicals in cell-free solutions and also in the surviving and intact suspended MGH-U1 cells by hydroethidine measurements. Under electron microscopical inspection, the same cells exhibited perinuclear cisternae, damaged mitochondria and numerous intracellular vacuoles. The contribution of free radicals to cell damage was investigated by reducing the vitamin E level in rats by a tocopherol free diet and by incubating L1210 cells in a tocopherol enriched medium. After 250 shock waves, ex vivo erythrocytes revealed a 75% increase in total cell disruption over cells from non-depleted rats. The in vitro experiments with L1210 cells exhibited a moderate protection by the addition of this scavenger of free radicals.Item Open Access Metabolism of 2-chloro-4-methylphenoxyacetate by Alcaligenes eutrophus JMP 134(1993) Pieper, Dietmar Helmut; Stadler-Fritzsche, Karin; Engesser, Karl-Heinrich; Knackmuss, Hans-Joachim2-Chloro-4-methylphenoxyacetate is not a growth substrate for Alcaligenes eutrophus JMP 134 and JMP 1341. It is, however, being transformed by enzymes of 2,4-dichlorophenoxyacetic acid metabolism to 2-chloro-4-methyl-cis, cis-muconate, which is converted by enzymatic 1,4-cycloisomerization to 4-carboxymethyl-2-chloro-4-methylmuconolactone as a dead end metabolite. Chemically, only 3,6-cycloisomerization occurs, giving rise to both diastereomers of 4-carboxychloromethyl-3-methylbut-2-en-4-olide. Those lactones harbonring a chlorosubstituent on the 4-carboxymethyl side chain were surprisingly stable under physiological as well as acidic conditions.Item Open Access Transduction of chemical signals in dictyostelium cells(1984) Gerisch, Günther; Tsiomenko, Arnold; Stadler, Joachim; Claviez, Michael; Hülser, Dieter F.; Rossier, ClaudeThree different functions of cyclic AMP in D discoideum are known: (1) cAMP acts as a chemoattractant during cell aggregation, (2) it controls cell development, particularly the acquisition of aggregation competence, and (3) it is involved in terminal cell differentiation. In this report we will concentrate on the functions 1 and 2 of cAMP. Chemotaxis requires the recognition of concentration gradients in the environment by attractant binding to cell surface receptors, the processing of signals from the receptors to the contractile system of the cells, extension of pseudopods at one part, and contraction at other parts of the cells in accord with the external gradient. One pathway of signal processing from the receptors to the contractile system involves the regulation of a myosin kinase. The control of development up to aggregation competence is largely dependent on the temporal pattern of cAMP application: Only repetitive pulses enhance development. This effect has been studied using the expression of a membrane glycoprotein called contact site A as a differentiation marker.Item Open Access Cyclic-AMP reception and cell recognition in dictyostelium discoideum(1975) Gerisch, Günther; Malchow, Dieter; Huesgen, Adolfine; Nanjundiah, Vidyanand; Roos, Werner; Wick, Ursula; Hülser, Dieter F.Single cells of the slime mold, Dictyostelium discoideum, aggregate into a multicellular organism in response to cyclic AMP, which they detect by binding to cellsurface receptors. During the aggregation phase, two different responses to cyclic-AMP are observed. First, the cells orientate by chemotaxis towards the source of a concentration gradient which initially is a group of cells forming an aggregation center. Second, the cells relay pulses which are periodically generated by the centers.Item Open Access Arachidonovaja kislota obratimo blokiruet vysokopronicaemye mežkletočnye kontakty(1994) Hülser, Dieter F.; Zempel, Günther; Reuss, Bernhard; Suhr, Dierk; Sarovskaja, Julija J.; Murav'eva, O. V.; Dunina-Barkovskaja, Antonina; Margolis, Leonid B.The effect of arachidonic on intercellular coupling via gap junctions has been studied in BICR/M1R k cells - a mammary tumor cell line of the Marshall ratt. Arachidonic acid is shown to reversibly block both ionic and dye coupling in a dose-dependent manner. The cells recoupled after the washout with either serum- or albumin (essentially fatty acid-free)-containing solution. The intercellular pH decreased from 7,2 to 7,0 after arachidonic acid treatment; the same pH shift in the absence of arachidonic acid, however, had no effect on the junctional permeability. Flow cytometric measurments revealed an arachidonic acid-induced increase of the cytoplasmic free Ca 2+ concentration which was also reversible upon albumin treatment. Intracellular Ca 2+ or H+ are unlikely to be involved in the mechanism of the arachidonic acid effect on intercellular coupling: high resolution measurments using double whole-cell technique also show reversible blockage of the junctional conductance in the presence of arachidonic acid while the pipette solution was buffered with 10 mM HEPES and 10 mM EGTA to clamp intracellular calcium and proton concentrations. We suggest that arachiconic acid directly affects the gap junction channels, probably interfering with the lipid-protein interactions.Item Open Access Pulsed high-power-sonication of concrements, cancer cells and rodent-tumors in vivo(1989) Riedlinger, Rainer E.; Brümmer, Franz; Hülser, Dieter F.Extracorporeal lithotripsy has been successfully established, based on different principles of generating and focusing the shock waves. Lithotripters have also been used to investigate the influence of shocks to cancer cells and solid tumors. With two different trans-mitters (spark-gap type XL-1 and piezo-resonance type MW 2) we applied shock waves and short high power US-pulses to suspended and immobilized tumor cells and multicell spheroids. With MW 2 significant local damage on cell spheroids in gelatin was achieved, caused by locally controlled cavitation. The results are compared to each other.Item Open Access Characteristics of three nuclear emulsions for autoradiography at the electron microscope(1966) Hülser, Dieter F.; Rajewsky, Manfred F.Drei handelsübliche Kernspuremulsionen, Gevaert NUC 307, Ilford L4 und Kodak NTE, wurden wegen ihrer geringen Korngröße auf ihre Eignung zur elektronenmikroskopischen Autoradiographie untersucht. Korngrößenverteilungskurven wurden aufgenommen und ein geeigneter Entwickler ausgesucht. Zur Bestimmung der Empfindlichkeit dieser drei Emulsionen wurden Einkornschichten im Elektronenmikroskop mit Elektronen einer Energie von 5,7 keV, der mittleren beta-Energie des Tritiums, bestrahlt. Anschließend wurden die Emulsionen entwickelt, aber nicht fixiert. Mit dem Anteil der entwickelten AgBr-Körner kann dann über Trefferkurven die Empfindlichkeit der Emulsionen bestimmt werden. Man benötigt zur Bildung eines latenten Bildkeimes für die Ilford L4-Emulsion 1 - 1,4 Elektronen pro AgBr-Korn, für die Gevaert NUC 307-Emulsion 2 - 3 und für die Kodak NTE-Emulsion 4 - 5 Elektronen pro AgBr-Korn. Folgerungen für das Auflösevermögen bei radioaktiven Punkt- und Flächenquellen werden diskutiert. Fortschritte in der Mikroautoradiographie werden von der Entwicklung feinkörniger Emulsionen abhängen, deren Empfindlichkeit bei etwa einem Elektron pro AgBr-Korn liegen sollte.