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 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 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 Nervous-system-specific carcinogenesis by ethylnitrosourea in the rat: molecular and cellular aspects(1977) Rajewsky, Manfred F.; Augenlicht, Leonard H.; Biessmann, Harald; Goth, Regine; Hülser, Dieter F.; Laerum, Ole D.; Lomakina, L. Ya.A lead in the search for cellular determinants favoring neoplastic transformation may be provided by the pronounced tissue specificity of the oncogenic effect of certain carcinogens which do not require enzymatic metabolic activation, i.e., in cases where this specificity cannot be due to tissue differences in the activity of enzymes involved in the formation of the ultimate reactants. A carcinogen that fulfills this condition is the ethylating agent N-ethyl-N-nitrosourea (EtNU). Alkylation of nucleic acid constituents by N-nitroso compounds in relation to mutagenesis and carcinogenesis has received considerable attention recently.Item Open Access Molecular and cellular mechanisms in nervous system-specific carcinogenesis by N-ethyl-N-nitrosourea(1976) Rajewsky, Manfred F.; Goth, Regine; Laerum, Ole D.; Biessmann, Harald; Hülser, Dieter F.A single pulse of N-ethyl-N-nitrosourea (ENU), applied to BDIX rats during the perinatal age, specifically results in a high incidence of neuroectodermal neoplasms in the central and peripheral nervous system (NS). The pronounced sensitivity of the developing NS suggests a dependence of the carcinogenic effect on the proliferative and/or differentiative state of the target cells at the time of the ENU pulse. The specificity of ENU for the NS cannot be due to tissue variations in the degree of carcinogen-cell interactions, since the reactive, electrophilic ethyl cation is produced by rapid, nonenzymatic decomposition of ENU indiscriminately in all tissues. Correspondingly, the initial molar fractions of ethylated purine bases are similar in the DNA of "high-risk" (perinatal brain) and "low-risk" tissues (e.g., liver; adult brain). However, while the respective half lives in DNA of N7-ethylguanine and N3-ethyladenine show only minor differences for both types of tissues, the mutagenic ethylation product 06-ethylguanine is removed from brain DNA very much more slowly than from the DNA of other tissues. Together with their high rate of DNA replication during the perinatal age, the incapacity of rat brain cells for enzymatic elimination of 06-alkylguanine from their DNA could account for an increased probability of neoplastic conversion, and hence for the NS specificity of ENU in the rat.Item Open Access Patch-clamp measurements of gap-junction channels in cultured cells(1992) Hülser, Dieter F.; Eckert, Reiner; Zempel, Günther; Paschke, Dietmar; Dunina-Barkovskaja, AntoninaDirect intercellular communication in most tissues is made possible by proteinaceous pores called gap-junction channels. These channels bridge the extracellular gap between apposed cells and connect their intracellular compartments both electrically and metabolically. The extracellular parts of two hemichannels - the connexons - are linked thus forming a communicating gap-junction channel. A connexon is a hexamer of protein subunits which are members of the connexin family. Since connexin 32 (Cx32) was the first gap-junction channel protein to be sequenced from hepatocytes, it serves as a reference to which all other gap-junction proteins are compared. The individual channel conductance may vary between 25 and 150 pS. Gap-junction channels of some tissues are more voltage sensitive (e.g. liver) than others (e.g. heart). The question whether these differences in electrical properties may be attributed to the different connexins being expressed in these tissues is still unanswered. Several approaches to resolve this problem will be discussed in this contribution, all are based on double whole-cell patch-clamp measurements using isolated cell pairs, as follows: (1) Cells with two different channel conductances perfused with anti connexin antibodies to specifically block one channel species; (2) Cells with only one connexin species selected by immunological characterization; (3) Weakly coupled HeLa cells transfected with specific connexin genes, a method which resulted in better correlations between connexin type and single channel properties.Item Open Access Investigations of different types of AgBr-layers for use in electron microscope autoradiography(1966) Hülser, Dieter F.; Rajewsky, Manfred F.On account of the low grain size three commercial emulsions GEVAERT NUC 307, ILFORD L4 and KODAK NTE have been investigated to assess their qualities for electron microscope microautoradiography. Grain size distribution curves were determined and a developer suitable for microautoradiography was selected after having tested different types of developers. In order to investigate the sensitivities of the three emulsions monolayer preparations were irradiated in the electron microscope using an energy of 5.7 keV, corresponding to the mean β-energy of Tritium. After exposure the specimens were developed but left unfixed. The sensitivity may then be determined, using the ratio of developed to the total number of grains. For the formation of one latent image the ILFORD L4 emulsion must be hit on the average by 1-1.4 electrons per AgBr-grain; the corresponding figures for GEVAERT NUC 307 and KODAK NTE are 2-3 and 4-5, respectively. The problem of resolution of point and plane sources of radioactivity is discussed.Item Open Access Shock waves and free radicals : cell protection by vitamin E in vitro and ex vivo(1993) Suhr, Dierk; Brümmer, Franz; Irmer, Ulrich; Schlachter, Manfred; Hülser, Dieter F.The application of extracorporeal generated shock waves in medicine for the fragmentation of human kidney and gall stones proved to be a very successful technique. Shock wave lithotripsy, however, is not free of tissue damaging side effects. One major mechanism for the fragmentation of stones as well as for the side effects is cavitation, ie. the formation and movement of bubbles in liquids exposed to tensile forces. Collapse of cavitation bubbIes is accompanied by local "hot spots" of several 1,000 K, thus generating free radicals. We investigated the contribution of these free radicals to cellular injury by varying the cellular amount of a well known scavenger of free radicals, α-tocopherol.