Measurement of the in-medium K0 inclusive cross section in pi(-) -induced reactions at 1.15 GeV/c.

The K0 meson production by pi(-) mesons of 1.15 GeV/c momentum on C, Al, Cu, Sn, and Pb nuclear targets was measured with the FOPI spectrometer at the Schwer-Ionen-Synchrotron accelerator of GSI. Inclusive production cross sections and the momentum distributions of K0 mesons are compared to scaled elementary production cross sections and to predictions of theoretical models describing the in-medium production of kaons. The data represent a new reference for those models, which are widely used for interpretation of the strangeness production in heavy-ion collisions. The presented results demonstrate the sensitivity of the kaon production to the reaction amplitudes inside nuclei and point to the existence of a repulsive KN potential of 20+/-5 MeV at normal nuclear matter density.

Nuclear stopping from 0.09A to 1.93A GeV and its correlation to flow.

We present a complete systematics (excitation functions and system-size dependences) of global stopping and side flow for heavy ion reactions in the energy range between 0.09A and 1.93A GeV. For the heaviest system, Au+Au, we observe a plateau of maximal stopping extending from about 0.2A to 0.8A GeV with a fast drop on both sides. The degree of stopping, which is shown to remain significantly below the expectations of a full stopping scenario, is found to be highly correlated to the amount of side flow.

Azimuthal dependence of collective expansion for symmetric heavy-ion collisions.

Detailed studies of the azimuthal dependence of the mean fragment and flow energies in the Au+Au and Xe+CsI systems are reported as a function of incident energy and centrality. Comparisons between data and model calculations show that the flow energy values along different azimuthal directions could be viewed as snapshots of the fireball expansion with different exposure times. For the same number of participating nucleons more transversally elongated participant shapes from the heavier system produce less collective transverse energy. Good agreement with Boltzmann-Uehling-Uhlenbeck calculations is obtained for a soft nuclear equation of state.

Free-radical-induced chain breakage and depolymerization of poly(methacrylic acid): equilibrium polymerization in aqueous solution at room temperature.

Hydroxyl radicals, generated by ionizing radiation in N2O saturated aqueous solutions, abstract H atoms from poly(methacrylic acid) at the methyl and methylene groups, and radicals 1 and 2 are formed, respectively. The reactions of the poly(methacrylic acid) radicals were investigated by pulse radiolysis (using optical and conductometric detection), EPR, product analysis, and kinetic simulations. The conductometric detection allowed us to measure the rate of chain scission and monomer release. Under conditions in which the polymer is largely deprotonated, the primary radical 1 abstracts a hydrogen (k= 3.5 x 10(2)s(-1)) from the methylene group, and this yields the more stable secondary radical 2. This radical undergoes chain scission by beta-fragmentation (k= 1.8 s(-1)), and the terminal (end-of-chain) radical 3 is formed. The polymer radicals terminate only slowly (2k= 80 dm3mol(-1)s(-1)). This allows effective depolymerization (depropagation) to take place (k=0.1 s(-1)). The yield of monomer release is higher than the original radical yield by up to two orders of magnitude. Once monomer is formed, it reacts with 3 (propagation, k= 15 dm3mol(-1)s(-1)), and a situation close to an equilibrium radical polymerization is approached. From these data, the equilibrium monomer concentration is calculated at 6.7 x 10(-3) mol dm(-3) at room temperature. The standard entropy of propagation is estimated at -185 to -150 J mol(-1)K(-1). Because the monomer reaches concentrations in the millimolar range, the *OH radicals increasingly react with monomers (results in oligomerization) rather than with the polymer. This effect is reflected by, for example, a lowering of chain-scission yields upon prolonged irradiation. In acid solutions, the decay of the polymer radicals becomes much faster (estimated at about 10(7)dm3mol(-1)s(-1) at pH3.5), and monomer release is no longer observed.

Time-resolved EPR studies on the reaction rates of peroxyl radicals of poly(acrylic acid) and of calf thymus DNA with glutathione. Re-examination of a rate constant for DNA.

Electron paramagnetic resonance (EPR) signals of peroxyl radicals of poly(acrylic acid), PAA, and of single- and double-stranded DNA were generated at 293 K and pH 6.6 by in situ photolysis of O2-saturated aqueous solutions containing the macromolecules and H2O2. From time-resolved EPR measurements upper limits for the rate constants of reaction of glutathione (GSH) with the peroxyl radicals of PAA [kb < or = (0.8 +/- 0.2) x 10(2) dm3 mol-1S-1] and of single- and double-stranded DNA from calf thymus [kb < or = (2 +/- 2) x 10(2) dm3 mol-1S-1] were determined. The low value of kb for reaction of GSH with DNA peroxyl radicals is in agreement with the observed lack of protective effect of the thiol in radiation-induced DNA strand break reactions (Liphard et al. 1990).

Phenoxyl radical complexes of gallium, scandium, iron and manganese.

The hexadentate macrocyclic ligands 1,4,7-tris(3,5-dimethyl-2-hydroxybenzyl)-1,4,7-triazacyclononane (L CH 3H3 ), 1,4,7-tris(3,5-di-tert-butyl-2-hydroxybenzyl)-1,4,7-triazacyclononane (L(Bu) H3 ) and 1,4,7-tris(3-tert-butyl-5-methoxy-2-hydroxybenzyl)-1,4,7-triazacyclononane (L OCH 3-H3 ) form very stable octahedral neutral complexes LM(III) with trivalent (or tetravalent) metal ions (Ga(III) , Sc(III) , Fe(III) , Mn(III) , Mn(IV) ). The following complexes have been synthesized: [L(Bu) M], where M = Ga (1), Sc (2), Fe (3); [L(Bu) Mn(IV) ]PF6 (4'); [L OCH 3M], where M = Ga (1 a), Sc (2 a), Fe (3 a); [L OCH 3Mn(IV) ]PF6 (4 a'); [L CH 3M], where M = Sc (2 b), Fe (3 b), Mn(III) (4 b); [L CH 3Mn(IV) ]2 (ClO4 )3 (H3 O)(H2 O)3 (4 b'). An electrochemical study has shown that complexes 1, 2, 3, 1 a, 2 a and 3 a each display three reversible, ligand-centred, one-electron oxidation steps. The salts [L OCH 3Fe(III) ]ClO4 and [L OCH 3Ga(III) ]ClO4 , have been isolated as stable crystalline materials. Electronic and EPR spectra prove that these oxidations produce species containing one, two or three coordinated phenoxyl radicals. The Mössbauer spectra of 3 a and [3 a](+) show conclusively that both compounds contain high-spin iron(III) central ions. Temperature-dependent magnetic susceptibility measurements reveal that 3 a has an S = 5/2 and [3a](+) an S = 2 ground state. The latter is attained through intramolecular antiferromagnetic exchange coupling between a high-spin iron(III) (S1 = 5/2) and a phenoxyl radical (S2 = 1/2) (H = - 2JS1 S2 ; J = - 80 cm(-1) ). The manganese complexes undergo metal- and ligand-centred redox processes, which were elucidated by spectroelectrochemistry; a phenoxyl radical Mn(IV) complex [Mn(IV) L OCH 3](2+) is accessible.

1H NMR studies of gamma-irradiated polynucleotides and DNA in N2O-saturated aqueous solutions: release of undamaged and modified bases.

In the 400-MHz 1H NMR spectra of gamma-irradiated, N2O-saturated aqueous solutions of polyuridylic acid (polyU), polycytidylic acid (polyC), polyadenylic acid (polyA), and single-stranded DNA from calf thymus, well-resolved signals of the undamaged free bases were identified. Upon incubation of the solutions at 95 degrees C for 2 h after irradiation, the amount of release of undamaged bases increased and in the case of polyA and DNA additional signals in the region 7.7-8.3 ppm due to 4,6-diamino-5-(formylamino)-pyrimidine and 8-hydroxyadenine (8-OH-adenine) were identified. Probably because of their low solubility guanine and modified guanine bases were not detected, whereas the lack of signals of low molecular weight pyrimidine derivatives is explained by the stability of the corresponding glycosidic bonds toward hydrolysis under our conditions. From the spectral intensities G values for release of unaltered and modified nucleic bases were estimated for radiation doses corresponding to approximately 2.5% conversion for polyU and polyC, to approximately 6% conversion for polyA and to approximately 30-60% conversion for DNA. The NMR data were in agreement with G values reported for release of undamaged bases from irradiated polyU and for double-strand DNA obtained on the basis of chromatographic procedures in combination with UV absorption (D.J. Deeble, D. Schulz, and C. von Sonntag, Int. J. Radiat. Biol. 49, 915-926, 1986; J.F. Ward and I. Kuo, Radiat. Res. 66, 485-498, 1976). Broadening of the NMR peaks of the polymers at high radiation doses (> 10 kGy) was dependent on the magnetic field strength. Therefore, it is ascribed to chemical shift heterogeneity caused by a variety of irradiation products with strongly overlapping signals rather than to a decrease in internal mobility caused by crosslinking of the polymer chains.

Intramolecular transformation reaction of the glutathione thiyl radical into a non-sulphur-centred radical: a pulse-radiolysis and EPR study.

The thiyl radical derived from glutathione (GSH) is shown to decay rapidly in aqueous solution by intramolecular rearrangement reactions into the non-sulphur-centred radical 1. The reaction is induced by OH- with a rate constant of 5 x 10(9) dm3 mol-1 and is also observable at near-neutral conditions (at physiological pH values around 7.5 the rate of formation of 1 amounts to approximately 1 x 10(3) s-1). The activation enthalpy and entropy at pH 8.4 and 20 degrees C were found to be 26.7 kJ mol-1 and -77 J mol-1 K-1, respectively. Radical 1 was unequivocally identified by EPR as the alpha-amino radical at the glutamyl residue of GSH. It is relatively long-lived with typical bimolecular decay rate constants of the order of (2-20) x 10(6) dm3 mol-1 s-1. At higher GSH concentrations the formation of 1 is retarded but not inhibited. All radicals, sulphur- as well as non-sulphur-centred ones are connected via equilibria, partly under the action of 'repair' processes of GSH. These repair processes, however, are slow (k much less than 1.4 x 10(5) dm3 mol-1 s-1). The equilibria are established quite rapidly and were found to be far on the side of the non-sulphur-centred radical under all conditions employed. Radical 1 possesses reducing properties as evidenced by its fast reaction with 4-nitro-acetophenone (PNAP) to yield PNAP.- (k = 7 x 10(8) dm3 mol-1 s-1).

The SO4-.-induced oxidation of 2'-deoxyuridine-5'-phosphate, uridine-5'-phosphate and thymidine-5'-phosphate. An ESR study in aqueous solution.

Reactions of photolytically generated SO4-. with 2'-deoxyuridine-5'-phosphate (5'-dUMP), uridine-5'-phosphate (5'-UMP) and thymidine-5'-phosphate (5'-dTUMP) were studied by ESR spectroscopy in aqueous solution under anoxic conditions. From 5'-dUMP and 5'-UMP the 5',5-cyclic phosphate-6-yl radicals 10 and 11 were generated (pH 2-11) whereas from 5'-dTMP at pH 3-8 the 5,6-dihydro-6-hydroxy-5-yl radical 14 and at pH 7-11 the 5-methylene-2'-deoxyuridine-5'-phosphate radical 15 was produced. In the experiments with 5'-UMP in addition to radical 11 the signals of sugar radicals 12 and 13 were detected. It is assumed that the base radical cations act as intermediates in the SO4-.-induced radical reactions. The 5'-phosphate group adds intramolecularly to the C(5)-C(6) bond of the uraclilyl radical cation whereas the thymidyl radical cation of 5'-dTMP reacts with H2O at pH less than 8 to yield the 6-OH-5-yl adduct 14 and deprotonates at pH greater than 7 thus forming the allyl-type radical 15. In 5'-UMP transfer of the radical site from the base to the sugar moiety competes with intramolecular phosphate addition.

ESR spectra of radicals of single-stranded and double-stranded DNA in aqueous solution. Implications for .OH-induced strand breakage.

In situ photolysis at 20 degrees C (argon plasma light source, lambda approximately greater than 200 mm) of oxygen-free solutions containing 2 mM H2O2 and heat-denatured, single-stranded (ss)DNA from calf-thymus resulted in the ESR spectra of the 6-hydroxy-5,6-dihydro-thymin-5-yl (1) and 5-methyleneuracil (3) radicals linked to the sugar-phosphate backbone. They were generated by reaction of OH radicals with DNA. By comparison of the decay characteristics of the ESR signals with rate constants from pulse-conductivity measurements [E. Bothe, G.A. Qureshi and D. Schulte-Frohlinde, Z. Naturforsch., 38c, 1030, (1983)] the thymine-derived radicals (1) and (3) can be excluded as precursors of the fast, dominating component of strand breakage of ssDNA. In the absence of H2O2 from native, double-stranded (ds)DNA an ESR signal was obtained (singlet, g approximately 2.004, delta v1/2 approximately 0.8 mT) which was assigned to the deprotonated guanine radical cation, [G.(-H)] of a DNA subunit. It is assumed that by the UV irradiation the guanine radical cation, (G+.), is generated, either by monophotonic photoionization or by electron transfer to pyrimidine bases. By rapid transfer of the bridging proton from (G+.) to the hydrogen bonded cytosine [G.(-H)] is formed. When photolysis of dsDNA was carried out in the presence of H2O2, reaction of photolytically generated .OH resulted in peroxyl radicals and purine radicals. The oxygen for formation of the peroxyl radicals is probably produced by reaction of [G.(-H)] with H2O2. Photolysis of N2O-saturated solutions containing dsDNA or ssDNA provided another possibility of generation of OH radicals. Under those conditions the .OH-induced radicals (1) and (3) were obtained not only from ssDNA but also from dsDNA.

E.s.r. studies on the mechanism of hydroxyl radical-induced strand breakage of polyuridylic acid.

Reaction of photolytically produced .OH radicals with polyuridylic acid [poly(U)] in neutral solutions resulted in the electron spin resonance (e.s.r.) spectrum of the C(5)-OH-6-yl (= 6-yl) radical 1b of the nucleobase. At pH less than or equal to 4 the spectrum of the base radical had disappeared and instead the cyclic 2'-oxo-3'-yl sugar radical 2a was observed. The assignment of the sugar radical was supported by model reactions with SO4-. as the radical inducing agent. Time-resolved e.s.r. measurements showed that the rate of decay of the 6-yl base radical at neutral pH is virtually the same as that of the strand break (sb) formation. These results prove that: (i) in agreement with an earlier proposal the C(2') mechanism contributes to sb formation of poly(U), and (ii) the decay of the 6-yl radical is the rate-determining step in the reaction sequence leading to strand breakage. The change in the e.s.r. spectra at pH 4 is due to an increase in the rate of sb formation with increasing proton concentration. This effect is explained by generation of the radical cation, 4, from the 6-yl radical and/or by rearrangement of the 6-yl radical into the C(6)-OH-5-yl (= 5-yl) radical in proton-induced reactions and subsequent rapid H abstraction from the sugar moieties by species 4 and/or 5.

Lysozyme-induced fusion of liposomes with erythrocyte ghosts at acidic pH.

Lysozyme that was covalently bound to the outer surface of sonicated vesicles induced fusion of the vesicles with human white erythrocyte ghosts. The kinetics of membrane mixing were evaluated by the resonance-energy-transfer method using L-alpha-dipalmitoyl phosphatidylethanolamine labeled at the free amino group with the energy donor 7-nitro-2,1,3-benzoxadiazol-4-yl or with the energy acceptor tetramethylrhodamine. The equilibrium state after fusion was characterized by using fluorescence photobleaching and recovery techniques. Rates and equilibrium percentages of fusion were maximal at the pH optimum of the enzyme, and rates were strongly reduced by the addition of N,N',N''-triacetylchitotriose, a competitive inhibitor of lysozyme. An apparent activation energy of 28 +/- kcal/mol was obtained for the lipid-mixing process. At 37 degrees C, the fusion half-time was 0.5 min. After 30 min at 37 degrees C, 40% of the labeled lipids initially present in the fusion mixture had a lateral diffusion constant, D, of 1.1 +/- 0.5 X 10(-9) cm2 X sec-1 in the ghost membrane. The strong induction of fusion at the lysozyme pH optimum was not observed in the absence of lysozyme or when free lysozyme was added to the solution. Bound lysozyme did not induce fusion of electrically neutral liposomes with each other. These observations indicate that it is the liposome-bound lysozyme that induces fusion between liposomes and erythrocyte ghosts.

N-NBD-L-alpha-dilauroylphosphatidylethanolamine. A new fluorescent probe to study spontaneous lipid transfer.

Migration of the fluorescent phospholipid N-(7-nitro-2,1,3-benzoxadiazol-4-yl)-L-alpha-dilauroylphosphati dylethanolamine between small sonicated egg phosphatidylcholine vesicles was studied by use of the fluorescence resonance energy transfer method. Contrary to the results of lipid transfer experiments reported for acyl chain NBD-labeled phospholipids (Nichols, J.W. and Pagano, R.E. (1982) Biochemistry 21, 1720-1728), the migration kinetics of N-NBD-DLPE had to be described by a sum of two exponential functions. The fast component (t1/2 approximately equal to 38 min) was assigned to lipid transfer via soluble monomers and the slow component (t1/2 approximately equal to 400 min) to transbilayer motion. A reversible four-stage process is suggested as a kinetic model. Mathematical treatment of this scheme is given yielding an analytical expression for the time dependence of NBD emission intensity. The use of N-NBD-DLPE in the resonance energy transfer measurements offers the advantage of simple chemical synthesis of the fluorescent probe and leads to additional information on transbilayer motion which was not available with the NBD-labeled lipids used so far.

Sugar transport by the bacterial phosphotransferase system. Nanosecond fluorescence studies of the phosphocarrier protein (HPr) labeled at the NH2-terminal methionine.

HPr is a low molecular weight, phosphocarrier protein of the Salmonella typhimurium phosphoenolpyruvate:glycose phosphotransferase system (PTS). This protein was alkylated with the fluorescent reagent (N-iodoacetylaminoethyl)-5-naphthylamino-1-sulfonate under conditions which favor alkylation of the thioether linkage in methionine residues (Link, T. P. and Stark, G. R. (1968) J. Biol. Chem. 243, 1082-1088) to give the corresponding sulfonium derivatives. The isolated fluorescent protein (95-100% pure) was as active as native HPr both as a phosphoryl acceptor protein (phosphoenolpyruvate and Enzyme I of the PTS), and as a phosphocarrier protein in the phosphorylation of methyl alpha-glucoside by the complete PTS. The fluorescent label was shown to be predominantly, possibly exclusively, at the NH2-terminal methionine residue. The decay of the fluorescence intensity could be described in terms of a biexponential function with the time constants tau 1 approximately 7 ns, tau 2 approximately 15 ns, and a ratio of alpha 2/alpha 1 approximately 3 for the pre-exponential factors. The decay of the fluorescence emission anisotropy was found to be consistent with some internal motion of the probe, in addition to the rotation of the protein conjugate as a whole.

[(Informed consent in clinical trials and its impact on methods of randomization and statistical evaluation)]. / Berücksichtigung der Aufklärungspflicht bei Therapiestudien und ihre Auswirkung auf Randomisierungs- und Auswertungsverfahren.

If patients eligible for a clinical trial are fully informed on alternative therapies, randomization, risks, benefits and right to refuse some of them will disagree with randomization. Random procedures which pay regard to that fact are described. The effect on statistical evaluation and sample sizes is investigated.

Nanosecond fluorescence anisotropy decays of 1,6-diphenyl-1,3,5-hexatriene in membranes.

Nanosecond decays of the fluorescence anisotropy, r, were studied for the emission of 1,6-diphenyl-1,3,5-hexatriene (DPH) embedded in a series of mixed multilamellar liposomes containing egg yolk phosphatidylcholine, phosphatidylethanolamine and cholesterol in varying molar ratios, as well as in membranes of intact cells and in virus envelopes. The relative contributions of the fast and the infinitely slow decaying component to the steady-state value r, of the fluorescence anisotropy were very similar for artifical and biological membranes. Angles, theta, of the cone, by which the motion of the fluorescent molecule is limited, were calculated from the intensity of the infinitely slow decaying anisotropy component and compared with steady-state fluorescence anisotropies and with 'microviscosities', (eta). An increase in (eta) from 1.5 to 5.2 P in our systems was accompanied by a decrease in theta from 49 degrees to 30 degrees while the decrease in the mean motional relaxation times, phi f, of the label molecule was not more than 1 ns and due mainly to changes in the potential, by which the diffusion of DPH in the membrane is restricted. From these observations we conclude that differences in the steady-state fluorescence anisotropy and in 'microviscosities' of cholesterol-containing membranes (r greater than 0.15) represent changes in the degree of static orientational constraint rather than changes in diffusion rates of the label.

Early molecular events in the interaction of enveloped viruses with cells. I. A fluorescence and radioactivity study.

The fluorescence depolarization of 1,6-diphenyl-hexatriene was used to study the dynamic properties of the hydrophobic regions of the lipid envelopes of ortho- and paramyxoviruses as well as of the Rous sarcoma virus and of the membrane lipids of susceptible and nonsusceptible cells. The systems investigated where active and inactive influenza viruses, and NDV virus acting on chick embryo fibroblasts and Rous sarcoma virus acting on susceptible (C/E) and nonsusceptible (C/B) chicken-cell. Polarization degrees and mean rotational correlation times of DPH embedded in viral lipids were significantly higher than those of DPH in the cell membranes, due to a higher rigidity of the virus envelopes. When suspensions of labelled viruses and unlabelled cells or unlabelled viruses and labelled cells were mixed, a characteristic change of the fluorescence polarization degrees with time was observed. This behaviour was ascribed to label transfer from virus to cell membranes or vice versa. While the rate constants of label transfer from virus to cells and cells to virus were about the same for the penetrating viruses the rate constants of label release from inactive virus to cells were much larger than for the migration in the opposite direction.