Showing papers by "University of Konstanz published in 1992"

A T cell-dependent experimental liver injury in mice inducible by concanavalin A.

Abstract: Male NMRI or BALB/c mice developed severe liver injury as assessed by transaminase release within 8 h when an intravenous dose greater than 1.5 mg/kg concanavalin A (Con A) was given. Histopathologically, only the liver was affected. Electron micrographs revealed leukocyte sticking to endothelial cells and bleb formation of hepatocytes. The hepatotoxicity of the lectin correlated neither with its agglutination activity nor with its sugar specificity. Administration of 0.5 mg/kg dexamethasone or 50 mg/kg cyclosporine A or 50 mg/kg FK 506 (Fujimycin) resulted in protection of the animals whereas indomethacin pretreatment failed to protect. Con A hepatitis was accompanied by the release of IL-2 into the serum of the animals. Mice with severe combined immunodeficiency syndrome lacking B as well as T lymphocytes were resistant against Con A. Athymic nude mice with immature T lymphocytes were also resistant. Pretreatment of mice with an antibody against T lymphocytes fully protected against Con A as did monoclonal anti-mouse CD4. Monoclonal anti-mouse CD8 failed to protect. Pretreatment of mice with silica particles, i.e., deletion of macrophages, prevented the induction of hepatitis. These findings provide evidence that Con A-induced liver injury depends on the activation of T lymphocytes by macrophages in the presence of Con A. The model might allow the study of the pathophysiology of immunologically mediated hepatic disorders such as autoimmune chronic active hepatitis.

The complete DNA sequence of yeast chromosome III.

Abstract: The entire DNA sequence of chromosome III of the yeast Saccharomyces cerevisiae has been determined. This is the first complete sequence analysis of an entire chromosome from any organism. The 315-kilobase sequence reveals 182 open reading frames for proteins longer than 100 amino acids, of which 37 correspond to known genes and 29 more show some similarity to sequences in databases. Of 55 new open reading frames analysed by gene disruption, three are essential genes; of 42 non-essential genes that were tested, 14 show some discernible effect on phenotype and the remaining 28 have no overt function.

Systematic study of insulator-metal transitions in perovskites RNiO3 (R=Pr,Nd,Sm,Eu) due to closing of charge-transfer gap.

Abstract: The detailed behavior of the phase transitions was mapped out for the series R${\mathrm{NiO}}_{3}$ as a function of the rare earth (R). A sharp insulator-metal transition is observed, which depends strongly on R.Forsmall$R it occurs at a higher temperature than the antiferromagnetic ordering (measured by muon-spin relaxation). By increasing either the temperature or the size of R, an insulator-metal transition is observed, most probably caused by the closing of the charge-transfer gap, induced by increase in the electronic bandwidth.

Dimensions and types of policy networks

Abstract: The paper offers a refined and systematic concept of state-business relations based on the 'policy network' idea. The major dimensions of policy networks are presented as (1) number and type of actors, (2) function of networks, (3) structure, (4) institutionalization, (5) rules of conduct, (6) power relations, (7) actor strategies. Certain popular conventional policy making arrangements (e.g. sectoral corporation, sponsored pluralism, clientelism) are examined in terms of the network dimensions.

Kinetics of CH4 oxidation in oxic soils exposed to ambient air or high CH4 mixing ratios

Abstract: The kinetic parameters of CH4 oxidation (Km, Vmax, apparent threshold = Tha) were measured using different oxic soils (cultivated cambisol, forest luvisol, meadow cambisol, paddy soil) both in a fresh state and after 3 weeks preincubation under high CH4 mixing ratios (20%). The preincubation resulted in an increase of the most probable number of methanotrophic bacteria. In fresh soils, CH4 oxidation followed Michaelis-Menten kinetics with a low Km (30–51 nM CH4), low Vmax (0.7–3.6 nmol CH4 h−1g−1dw soil), and low Tha (0.2–2.7 ppmv CH4). In preincubated soils, CH4 oxidation exhibited biphasic kinetics in which two different CH4 saturation curves were apparently superimposed on each other. Eadie-Hofstee plots of the data showed two activities with different kinetic parameters: a high-affinity activity with low Km (13–470 nM CH4), low Vmax (2.1–150.0 nmol CH4 h−1g−1dw) and low Tha (0.3–4.1 ppmv CH4) being similar to the kinetic parameters in fresh soils; and a low-affinity activity with high Km (1740–27 900 nM CH4), high Vmax (270–3 690 nmol CH4 h−1g−1dw) and high Tha (11–45 ppmv CH4) being similar to the kinetic parameters known from methanotrophic bacteria. The low-affinity activity was also observed in a soil over a deep natural gas source which was permanently exposed to high CH4 mizing ratios (>5% CH4). Bacteria culturable as methanotrophs are probably responsible for the low-affinity activity which is typical for the soils exposed to high CH4 mixing ratios. However, the bacteria responsible for the high-affinity activity are still unknown. This activity is typical for the soils exposed to only ambient CH4 mixing ratios. Both high- and low-affinity activities were inhibited by autoclaving and by acetylene.

An extremely low‐light adapted phototrophic sulfur bacterium from the Black Sea

Abstract: Five strains of a brown phototrophic sulfur bacterium (Chlorobium phaeobacteroides) were isolated from the chemocline of the Black Sea (80-m depth). All contain bacteriochlorophyll e as the main photosynthetic pigment. The strains revealed extreme low-light adaptation of growth compared to 12 other green and purple sulfur bacterial strains. At very low light intensities (<4 µEinst m‒2 s‒1), the Black Sea strain MN 1 oxidized sulfide faster than the type strain 2430; the latter reached three times higher oxidation rates at light saturation. Low-light adaptation is achieved by an increase of light-harvesting pigments (175% compared to the type strain) and a very low maintenance energy requirement. The efficiency of energy transfer (59%) within light-harvesting structures (chlorosomes) is comparable in other green sulfur bacteria and, therefore, appears to be limited by the molecular organization of the chlorosomes. From data in the literature, a light transmission of 0.0005% of surface irradiance was calculated for the chemocline of the Black Sea. Extrapolation of our laboratory data revealed that anoxygenic photosynthesis could account for 4% of total sulfide oxidation under average light conditions in situ and for 13% at maximal surface irradiance in summer.

Oxidation of H2, organic compounds and inorganic sulfur compounds coupled to reduction of O2 or nitrate by sulfate-reducing bacteria

Abstract: All of fourteen sulfate-reducing bacteria tested were able to carry out aerobic respiration with at least one of the following electron donors: H2, lactate, pyruvate, formate, acetate, butyrate, ethanol, sulfide, thiosulfate, sulfite. Generally, we did not obtain growth with O2 as electron acceptor. The bacteria were microaerophilic, since the respiration rates increased with decreasing O2 concentrations or ceased after repeated O2 additions. The amounts of O2 consumed indicated that the organic substrates were oxidized incompletely to acetate; only Desulfobacter postgatei oxidized acetate with O2 completely to CO2. Many of the strains oxidized sulfite (completely to sulfate) or sulfide (incompletely, except Desulfobulbus propionicus); thiosulfate was oxidized only by strains of Desulfovibrio desulfuricans; trithionate and tetrathionate were not oxidized by any of the strains. With Desulfovibrio desulfuricans CSN and Desulfobulbus propionicus the oxidation of inorganic sulfur compounds was characterized in detail. D. desulfuricans formed sulfate during oxidation of sulfite, thiosulfate or elemental sulfur prepared from polysulfide. D. propionicus oxidized sulfite and sulfide to sulfate, and elemental sulfur mainly to thiosulfate. A novel pathway that couples the sulfur and nitrogen cycles was detected: D. desulfuricans and (only with nitrite) D. propionicus were able to completely oxidize sulfide coupled to the reduction of nitrate or nitrite to ammonia. Cell-free extracts of both strains did not oxidize sulfide or thiosulfate, but formed ATP during oxidation of sulfite (37 nmol per 100 nmol sulfite). This, and the effects of AMP, pyrophosphate and molybdate on sulfite oxidation, suggested that sulfate is formed via the (reversed) sulfate activation pathway (involving APS reductase and ATP sulfurylase). Thiosulfate oxidation with O2 probably required a reductive first step, since it was obtained only with energized intact cells.

Granulocyte colony-stimulating factor treatment protects rodents against lipopolysaccharide-induced toxicity via suppression of systemic tumor necrosis factor-alpha.

Abstract: Pretreatment with recombinant human granulocyte CSF (G-CSF) protected mice in two different models of septic shock. Intravenous injection of 250 micrograms/kg G-CSF to mice prevented lethality induced by 5 mg/kg LPS. Injection of 50 micrograms/kg G-CSF protected galactosamine-sensitized mice against LPS-induced hepatitis. In either case, this protection was accompanied by a suppression of LPS-induced serum TNF activity. In contrast, when galactosamine-sensitized mice were pretreated with 50 micrograms/kg murine recombinant granulocyte/macrophage CSF instead of G-CSF and subsequently challenged with LPS, serum TNF activity was significantly enhanced and mortality was increased. The suppressive effect of G-CSF on LPS-induced TNF production was also demonstrated in rats. In vivo, no TNF was detectable in the blood of LPS-treated rats, which had been pretreated with G-CSF. Ex vivo, alveolar macrophages, bone marrow macrophages, Kupffer cells, or peritoneal macrophages prepared from G-CSF-treated rats produced significantly less TNF upon stimulation with LPS than corresponding populations from control rats. However, when these macrophage populations were incubated with G-CSF in vitro, LPS-induced TNF production was unaffected. These data suggest that the G-CSF-mediated suppression of TNF production is not a direct effect of G-CSF on macrophages. To examine whether, independent of the protection against LPS, G-CSF treatment still activated neutrophils, it was demonstrated that granulocytes from G-CSF-treated rats were primed for PMA-induced oxidative burst and for ionophore/arachidonic acid-stimulated lipoxygenase product formation. The experiments of this study support the notion that G-CSF is a negative feedback signal for macrophage-derived TNF-alpha production during Gram-negative sepsis.

Characterization of SAF-A, a novel nuclear DNA binding protein from HeLa cells with high affinity for nuclear matrix/scaffold attachment DNA elements.

Abstract: We identified four proteins in nuclear extracts from HeLa cells which specifically bind to a scaffold attachment region (SAR) element from the human genome. Of these four proteins, SAF-A (scaffold attachment factor A), shows the highest affinity for several homologous and heterologous SAR elements from vertebrate cells. SAF-A is an abundant nuclear protein and a constituent of the nuclear matrix and scaffold. The homogeneously purified protein is a novel double stranded DNA binding protein with an apparent molecular weight of 120 kDa. SAF-A binds at multiple sites to the human SAR element; competition studies with synthetic polynucleotides indicate that these sites most probably reside in the multitude of A/T-stretches which are distributed throughout this element. In addition we show by electron microscopy that the protein forms large aggregates and mediates the formation of looped DNA structures.

Simian virus 40 T-antigen DNA helicase is a hexamer which forms a binary complex during bidirectional unwinding from the viral origin of DNA replication.

Abstract: The role of simian virus 40 (SV40) large tumor antigen (T antigen) as a DNA helicase at the replication fork was studied. We found that a T-antigen hexamer complex acts during the unidirectional unwinding of appropriate DNA substrates and is localized directly in the center of the fork, contacting the adjacent double strand as well as the emerging single strands. When bidirectional DNA unwinding, initiated at the viral origin of DNA replication, was analyzed, a larger T-antigen complex that is simultaneously active at both branch points of an unwinding bubble was observed. The size and shape of this helicase complex imply that the T-antigen dodecamer complex, assembled at the origin and active in the localized melting of duplex DNA, is subsequently also used to continue DNA unwinding bidirectionally. Then, however, the dodecamer complex does not split into two hexamer subunits that track along the DNA; rather, the DNA is threaded through the intact complex, with the concomitant extrusion of single-stranded loops.

Adhesion Pad Formation and the Involvement of Cutinase and Esterases in the Attachment of Uredospores to the Host Cuticle.

Abstract: We have investigated the basis of adhesion of uredospores of the obligately parasitic rust fungus Uromyces viciae-fabae to leaves of its broad bean host. Upon contact with an aqueous environment, spores form a structure that we have termed an adhesion pad. The adhesion pad is formed by both living and autoclaved spores, but only adhesion pads formed by living spores adhered to the cuticle of leaves of the host plant. Treatment of living spores with the serine-esterase inhibitor diisopropyl fluorophosphate prevented the adhesion of the pad to the leaf surface, suggesting a functional role for esterase or cutinase in the process of adhesion. A cutinase and two nonspecific serine-esterases were found to be localized on the surface of spores. These enzymes were released rapidly from the spore surface upon contact with an aqueous environment. The addition of the cutinase and the nonspecific esterases to autoclaved spores restored their ability to adhere to the host cuticle. Thus, whereas pad formation appears to be a passive response to the aqueous environment, the actual adhesion of pads to the host cuticle appears to depend on the cutinase and esterases associated with the spore surface. These results suggest a new role for cutinases and serine-esterases in the fungal infection process.

Identification and molecular analysis of glgS, a novel growth-phase-regulated and rpoS-dependent gene involved in glycogen synthesis in Escherichia coli.

Abstract: The putative stationary-phase sigma factor (sigma S) encoded by rpoS is essential for glycogen synthesis, but is not required for the transcription of glgC and glgA, which encode ADP-glucose-pyrophosphorylase and glycogen synthase, respectively. Using a mini-Mu random chromosomal library and a screen for glycogen overproduction, we identified a novel gene (glgS) involved in glycogen synthesis. glgS maps at 66.6 min (3247 kb) on the chromosome and constitutes a monocistronic operon. It encodes a hydrophilic and highly charged small protein, with a molecular weight of 7886, which is strongly expressed in minicells. Experiments with single-copy chromosomal glgS::lacZ gene fusions indicated that glgS expression is controlled by sigma S as well as by cAMP. Two transcriptional start sites were mapped in the upstream regulatory region of glgS. The glgSp1 transcript was absent in a cya mutant, whereas an rpoS mutant did not synthesize the glgSp2 transcript. Although glycogen synthesis is strongly stimulated by overproduction of GlgS and is inhibited by a glgS null mutation, glgS does not affect the expression of the glgCAP operon. Its potential role in the metabolic control of glycogen synthesis is discussed. Also, evidence is presented to show that the amount of glycogen accumulated in vivo in early stationary-phase cells is mainly determined by sigma S-controlled gene expression and allosteric activation of GlgC, whereas the absolute levels of expression of glgCAP as well as the intracellular concentration of cAMP are of minor importance.

A comparative EPR investigation of the multicopper proteins nitrous‐oxide reductase and cytochrome c oxidase

Abstract: The multicopper proteins, nitrous-oxide reductase (N2OR) and cytochrome c oxidase (COX), were investigated by EPR spectroscopy at microwave frequencies 2.4-35 GHz. Our results support a Cu-Cu interaction in COX and N2OR. At least 10 lines in the 2.7-GHz, 12 lines in the 4.6-GHz and 14 lines in the 9.2 GHz spectra were resolved for N2OR. Eight copper lines at 2.7 GHz, about nine lines at 4.6 GHz and about six lines at 9.2 GHz were resolved for COX. Simulations of the EPR spectra were consistent with most of the resonances of the multiline spectra, including regions in the center of the spectra where overlap of the three seven-line patterns is proposed. These simulations indicated that Cu-Cu interaction, in a mixed-valence [Cu(1.5) ... Cu(1.5)], S = 1/2 site is consistent with, if not proof of, the unusual spectral features observed for N2OR and COX.

Trying to understand axonal regeneration in the CNS of fish.

Abstract: SUMMARY In contrast to the situation in mammals and birds, neurons in the central nervous system (CNS) of fish-such as the retinal ganglion cells-are capable of regenerating their axons and restoring vision. Special properties of the glial cells and the neurons of the fish visual pathway appear to contribute to the success of axonal regeneration. The fish oligodendrocytes lack the axon growth inhibiting molecules that interfere with axonal extension in mammals. Instead, fish optic nerve oligodendrocytes support-at least in vitro-axonal elongation of fish as well as that of rat retinal axons. Moreover, the fish retinal ganglion cells re-express upon injury a set of growthassociated cell surface molecules and equip the regenerating axons throughout their path and up into their target, the tectum opticum with these molecules. This may indicate that the injured fish ganglion cells reactivate the cellular machinery necessary for axonal regrowth and pathfinding. Furthermore, the target itself provides positional marker molecules even in adult fish. These marker molecules are required to guide the regenerating axons back to their retinotopic home territory within the tCCtUm.

Properties of the nuclear P1 protein, a mammalian homologue of the yeast Mcm3 replication protein.

Abstract: Polyclonal antibodies were raised against a multiprotein 'holoenzyme' form of calf thymus DNA polymerase alpha-primase and used to probe a human cDNA-protein expression library constructed in the lambda gt11 vector. The probe identified a series of cDNA clones derived from a 3.2 kb mRNA which encodes a novel 105 kDa polypeptide, the P1 protein. In intact cells, the P1 protein was specifically associated with the nucleus, and in cell extracts, it was associated with complex forms of DNA polymerase alpha-primase. The synthesis of human P1-specific mRNA was stimulated upon addition of fresh serum to growth-arrested cells, and RNA blot analyses with the human P1-cDNA probe indicated that P1 is encoded by a strictly conserved mammalian gene. The amino acid sequence deduced from a 240-codon open reading frame resident in the largest human P1-cDNA (0.84 kb) displayed greater than 96% identity with that deduced from the equivalent segment of a 795-codon open reading frame of a larger mouse P1-cDNA (2.8 kb). Throughout its length, the primary structure of mammalian P1 displayed strong homology with that of Mcm3, a 125 kDa yeast protein thought to be involved in the initiation of DNA replication (Gibson et al. 1990. Mol. Cell. Biol. 10: 5707-5720). The P1-Mcm3 homology, the strong conservation of P1 among mammals, its nuclear localization, and its association with the replication-specific DNA polymerase alpha strongly suggest an important role of the P1 protein in the replication of mammalian DNA.

Dynamics of terminal arbor formation and target approach of retinotectal axons in living zebrafish embryos: a time-lapse study of single axons

Abstract: In a variety of species, developing retinal axons branch initially more widely in their visual target centers and only gradually restrict their terminal arbors to smaller and defined territories. Retinotectal axons in fish, however, appeared to grow in a directed manner and to arborize only at their retinotopic target sites. To visualize the dynamics of retinal axon growth and arbor formation in fish, time-lapse recordings were made of individual retinal ganglion cell axons in the tectum in live zebrafish embryos. Axons were labeled with the fluorescent carbocyanine dyes Dil or DiO inserted as crystals into defined regions of the retina, viewed with 40x and 100x objectives with an SIT camera, and recorded, with exposure times of 200 msec at 30 or 60 sec intervals, over time periods of up to 13 hr. (1) Growth cones advanced rapidly, but the advance was punctuated by periods of rest. During the rest periods, the growth cones broadened and developed filopodia, but during extension they were more streamlined. (2) Growth cones traveled unerringly into the direction of their retinotopic targets without branching en route. At their target and only there, the axons began to form terminal arborizations, a process that involved the emission and retraction of numerous short side branches. The area that was permanently occupied or touched by transient branches of the terminal arbor--"the exploration field"--was small and almost circular and covered not more than 5.3% of the entire tectal surface area, but represented up to six times the size of the arbor at any one time. These findings are consistent with the idea that retinal axons are guided to their retinotopic target sites by sets of positional markers, with a graded distribution over the axes of the tectum.

Nitrite oxidoreductase from nitrobacter hamburgensis redox centers and their catalytic role

Abstract: Nitrite oxidoreductase was isolated from mixotrophically grown cells of Nitrobacter hamburgensis. The enzyme purified from heat treated membranes was homogeneous by the criteria of polyacrylamide gel electrophoresis and size exclusion chromatography. The monomeric form consisted of two subunits with Mr 115000 and 65000, respectively. The dimeric form of the enzyme contained 0.70 molybdenum, 23.0 iron, 1.76 zinc, and 0.89 copper. The catalytically active enzyme was investigated by visible and electron paramagnetic resonance spectroscopy (EPR) under oxidizing (as isolated), reducing (dithionite), and turnover (nitrite) conditions. As isolated the enzyme exhibited a complex set of EPR signals between 5–75 K, originating from several ironsulfur and molybdenum (V) centers. Addition of the substrate nitrite, or the reducing agent dithionite resulted in a set of new resonances. The molybdenum and the iron-sulfur centers of nitrite oxidoreductase from Nitrobacter hamburgensis were involved in the transformation of nitrite to nitrate.

Preferential belief change using generalized epistemic entrenchment

Abstract: A sentence A is epistemically less entrenched in a belief state K than a sentence B if and only if a person in belief state K who is forced to give up either A or B will give up A and hold on to B. This is the fundamental idea of epistemic entrenchment as introduced by Gardenfors (1988) and elaborated by Gardenfors and Makinson (1988). Another distinguishing feature of relations of epistemic entrenchment is that they permit particularly simple and elegant construction recipes for minimal changes of belief states. These relations, however, are required to satisfy rather demanding conditions. In the present paper we liberalize the concept of epistemic entrenchment by removing connectivity, minimality and maximality conditions. Correspondingly, we achieve a liberalization of the concept of rational belief change that does no longer presuppose the postulates of success and rational monotony. We show that the central results of Gardenfors and Makinson are preserved in our more flexible setting. Moreover, the generalized concept of epistemic entrenchment turns out to be applicable also to relational and iterated belief changes.

Generation of a vortex chain in the wake of a Suhundulatory swimmer

Abstract: For a fish swimming at constant speed thrust must be sufficient to overcome drag. Drag of an undulating fish can differ considerably from drag observed for a rigid body and cannot be measured directly. Thrust can be calculated by applying hydrodynamic models, such as slender body theory [1], to observed kinematic patterns. Due to simplifying assumptions, shortcomings in the various applications, and probably behavioral variability, the estimated thrust coefficients (CT) range from ca. 0.5 c D to ca. 5 CD [2, 3] for animals swimming at the same Reynolds numer (c D: drag coefficient). • We present an estimate of thrust and efficiency based for the first time on the flow in the wake of freely swimming rainbow trout (Oncorhynchus mykiss Waldbaum; cf. [4, 5]). This calculation required the reconstruction of the three-dimensional vortex pattern as well as a quantification of the flow velocities, both of which are missing in studies so far available [6, 7]. We