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Biomedical subjects

R Grunwald

Publications and source records attributed to R Grunwald.

15 recordsLinked to original sources
PubMed2004

Vacuum-ultraviolet beam array generation by flat micro-optical structures.

Micro-optical structures for VUV laser beam shaping and wave-front sensing were manufactured by thin-film deposition onto CaF2 and transfer by etching. Arrays of Bessel-like F2 laser beams at a wavelength of 157 nm with extremely small conical angles were generated by microaxicon lenses. Beam propagation was studied in simulations and experiments. Apodization by absorbing layers is proposed for beam cleaning.

Journal Article↗
PubMed2003

Ultrashort-pulse wave-front autocorrelation.

Combined spatially resolved collinear autocorrelation and Shack-Hartmann wave-front sensing of femtosecond laser pulses is demonstrated for the first time to our knowledge. The beam is divided into multiple nondiffracting subbeams by thin-film micro-optical arrays. With hybrid refractive-reflective silica/silver microaxicons, wave-front autocorrelation is performed in oblique-angle reflection. Simultaneous two-dimensional detection of local temporal structure and wave-front tilt of propagating few-cycle wave packets is demonstrated.

Journal Article↗
PubMed2001

Spatially resolved small-angle noncollinear interferometric autocorrelation of ultrashort pulses with microaxicon arrays.

Small-angle, noncollinear, first- and second-order interferometric autocorrelation experiments with Ti:sapphire laser pulses of 9-80-fs duration have been performed with microaxicon arrays. Predictions of short-pulse spatial frequency effects were verified by comparison of interference patterns of single elements and matrices. An angular spectrum of Gaussian-shaped axicons was analyzed on the basis of linear refraction. Experimental data indicate contributions to autocorrelation by nonlinear refraction and travel-time differences. The influence of the spectral bandwidth was separated from the pulse-duration-dependent effects. Spatially resolved information about the coherence time was delivered by the multichannel structure.

Journal Article↗
PubMed1995

Lumenal sites and C terminus accessibility of the skeletal muscle calcium release channel (ryanodine receptor).

The membrane topology of the skeletal muscle ryanodine receptor (RyR1) was investigated using site-directed antibodies directed against amino acid sequences 2804-2930, 4581-4640, 4860-4886, and 4941-5037. Ab(2804-2930) bound with identical affinity to either closed or permeabilized sarcoplasmic reticulum vesicles, confirming the cytoplasmic location of this segment. Ab(4581-4640) did not bind to closed vesicles but bound well to permeabilized vesicles, supporting a lumenal location for this segment. Ab(4860-4886) did not bind to closed vesicles but exhibited weak binding to the permeabilized vesicles, suggesting that a portion of the epitope may be exposed on the lumenal surface. The C-terminal antibody (Ab(4941-5037)) bound weakly to closed vesicles, and binding was not significantly enhanced by permeabilizing vesicles with low concentrations of non-denaturing detergent. However, the C-terminal antibodies bound efficiently to vesicles which were transiently incubated at alkaline pH or subjected to trypsinolysis, conditions where few of the vesicles were permeabilized. These results support a model for the membrane topology of the ryanodine receptor as proposed by Takeshima et al. (Takeshima, H., Nishimura, S., Matsumoto, T., Ishida, H., Kangawa, K., Minamino, N., Matsuo, H., Ueda, M., Hanaoka, M., Hirose, T., and Numa, S. (1989) Nature 339, 439-445). The results also suggest that the native conformation of the C terminus is inaccessible to antibodies.

Animals↗
PubMed1990

The effects of lipid composition on the binding of lasalocid A to small unilamellar vesicles.

The binding of the carboxylic ionophore lasalocid A (X537A) to small unilamellar phospholipid vesicles of varying composition was examined in an effort to determine what structural features of the phospholipid membrane influence the ionophore-membrane interaction. Apparent dissociation constants (Kapp) were calculated for both the acidic and anionic forms of the ionophore using the change in fluorescence intensity observed for lasalocid A upon addition of phospholipid vesicles. The Kapp for binding to fluid phase dimyristoylphosphatidylcholine (DMPC) vesicles is 46 microM for the anion and 14 microM for the acid. While the phase transition of DMPC had no effect on the Kapp of the anion, an increase was observed in the Kapp of the acid below the phase transition temperature. The Kapp of the anion was not affected by the incorporation of 10% dimyristoylphosphatidylethanolamine (DMPE), but increased slightly upon incorporation of cholesterol. The pKa values of the ionophore were the same in DMPC and DMPC/DMPE membranes. Incorporation of the negative lipids phosphatidylglycerol, phosphatidic acid, or phosphatidylethanolamine (at pH 9.4 where PE carries a negative charge) decreases binding of the anion in accord with the increase in surface potential estimated from Gouy-Chapman theory. The CD spectrum of membrane-bound lasalocid A anion indicated the ionophore to be in an extended acyclic conformation on the membrane surface with the C-1 carboxylate rotated out of the plane of the salicylate ring. The out-of-plane rotation of the carboxylate may be the result of facial binding by the amphiphilic ionophore on the membrane surface or of weak ion pairing to the polar lipid head groups. These results suggest that the primary determinants of binding of the anionic ionophore on the membrane surface are packing density of the polar head groups and membrane surface potential. There is no evidence of strong hydrogen bond formation between the lipid polar head groups and the ionophore as has previously been suggested.

Circular Dichroism↗
PubMed1988

Interaction of the antifolate antibiotic trimethoprim with phosphatidylcholine membranes: a 13C and 31P nuclear magnetic resonance study.

The interaction of the bacterial dihydrofolate reductase inhibitor trimethoprim with small unilamellar 1,2-dimyristoyl-sn-glycero-3-phosphorylcholine vesicles was studied using 13C and 31P NMR spectroscopy. In an effort to determine whether trimethoprim passively permeates the vesicle membrane, an impermeant, anionic complex of the paramagnetic ion Dy3+ was added to the extravesicular compartment. Based on the downfield shift that the Dy3+ complex induces in the [2-13C] resonance of trimethoprim in free solution, membrane permeation and movement of the drug into the intravesicular space can in principle be established from observation of the C2 chemical shift alone. In contrast to what is predicted by a two-compartment model separated by a semipermeable barrier, the presence of vesicles virtually reverses the effect of the shift reagent on the [2-13C] carbon resonance. These results suggest that the majority of the trimethoprim might be sequestered within the vesicle membrane. A saturable decrease in the spin-lattice relaxation time and a saturable increase in the line width at half-height of the [2-13C] resonance as a function of vesicle concentration indicated that trimethoprim does in fact bind to the phospholipid matrix of the membrane bilayer. The KD for the interaction calculated from the relaxation data was 9.7 +/- 0.3 X 10(-4) M at a pH of 7.01 and an ionic strength of 0.015 M. The chemical shift of the [2-13C] resonance is unaffected by interaction with the electroneutral membrane, and the pKa increases by only 0.16 upon binding. These results point to an interfacial location for the pyrimidine moiety. Using the paramagnetic shift reagent Pr3+ and the 31P NMR signal from the phosphodiester groups of the membrane lipids, trimethoprim was shown to displace Pr3+ ions from binding sites on the outer membrane surface as would be expected if the polar pyrimidine ring were located at or near the membrane surface. The extent to which trimethoprim and trimethoprim derivatives modified in the 3'- and 4'-positions interact with the exo face of the membrane is strongly dependent on the type of substituent and whether it is in the 3'- or 4'-position. Van der Waals interactions between the 5-benzyl sidechain and the hydrophobic fatty acid region of the membrane interior appear to be necessary for the polar portion of trimethoprim to compete favorably for the membrane-binding site with the polyvalent Pr3+ ion.(ABSTRACT TRUNCATED AT 400 WORDS)

Cell Membrane Permeability↗
PubMed1987

[Heart rate and visual evoked potentials during figuring arithmetic problems in boys acquired in early childhood with minimal brain damage].

Changes of the pattern reversal EP and heart rate (HR) during calculating were studied on 32 boys with minimal brain damage and a control group (n 27) matched in age, sex and intelligence. The introduction of a reference period and repetition of the calculation period should improve the value of all results and should give simultaneously a first insight in the course of the parameters investigated, during mental loading. Both groups reached the same results in calculating, yet only the brain damaged boys simultaneously showed significant longer latencies of the EP. This result was interpreted as an effect of the higher loading of the damaged subjects. The task-depended increase of the HR was significant in both groups. The HR-level was getting lower with the increasing number of brain damage criteria. The results of intercorrelation between in physiologic parameters and datas of subjective feeling and results of calculating was negative in all cases. These findings were interpreted on the basis of several difficulties to find out clear cut patterns of reaction for different clinical groups.

Arousal↗
PubMed1986

Rhodamine 123 as a probe of transmembrane potential in isolated rat-liver mitochondria: spectral and metabolic properties.

The spectral and metabolic properties of Rhodamine 123, a fluorescent cationic dye used to label mitochondria in living cells, were investigated in suspensions of isolated rat-liver mitochondria. A red shift of Rhodamine 123 absorbance and fluorescence occurred following mitochondrial energization. Fluorescence quenching of as much as 75% also occurred. The red shift and quenching varied linearly with the potassium diffusion potential, but did not respond to delta pH. These energy-linked changes were accompanied by dye uptake into the matrix space. Concentration ratios, in-to-out, approached 4000:1. A large fraction of internalized dye was bound. At concentrations higher than those needed to record these spectral changes, Rhodamine 123 inhibited ADP-stimulated (State 3) respiration of mitochondria (Ki = 12 microM) and ATPase activity of inverted inner membrane vesicles (Ki = 126 microM) and partially purified F1-ATPase (Ki = 177 microM). The smaller Ki for coupled mitochondria was accounted for by energy-dependent Rhodamine 123 uptake into the matrix. Above about 20 nmol/mg protein (10 microM), Rhodamine 123 caused rapid swelling of energized mitochondria. Effects on electron-transfer reactions and coupling were small or negligible even at the highest Rhodamine 123 concentrations employed. delta psi-dependent Rhodamine 123 uptake together with Rhodamine 123 binding account for the intense fluorescent staining of mitochondria in living cells. Inhibition of mitochondria ATPase likely accounts for the cytotoxicity of Rhodamine 123. At concentrations which do not inhibit mitochondrial function, Rhodamine 123 is a sensitive and specific probe of delta psi in isolated mitochondria.

Adenosine Diphosphate↗
PubMed1984

Thermodynamic limits to the ATP/site stoichiometries of oxidative phosphorylation by rat liver mitochondria.

From measurements of reactants, products, and the oxidation-reduction state of cytochrome c + c1 during 3-hydroxybutyrate-supported oxidative phosphorylation by rat liver mitochondria at static head (state 4), we determined the free energy change of ATP formation from ADP and Pi (phosphorylation potential or delta GP) and the oxidation-reduction free energy changes (redox potentials or delta GR values) across Sites 1 + 2 (delta GR1 + 2), across Site 3 (delta GR3), and across Sites 1 + 2 + 3 (delta GR). At pH 7.4, -delta GR1 + 2/delta GP, -delta GR3/delta GP, and -delta GR/delta GP were maximally 1.80, 1.56, and 3.37. These can be taken as thermodynamic upper limits to the ATP/Sites 1 + 2, ATP/Site 3, and ATP/O stoichiometry of 3-hydroxybutyrate-supported oxidative phosphorylation. The theory of linear nonequilibrium thermodynamics were employed to estimate lower limits to the ATP/site stoichiometries. The lower limit is given by the expression, q2(-delta GRsite/delta GP). The degree of coupling, q, was 0.977 as determined from the dependence of respiratory rate on delta GP. Determined in this way, lower limits of the ATP/Sites 1 + 2, ATP/Site 3, and ATP/O stoichiometries were 1.67, 1.44, and 3.11, respectively. ADP addition to mitochondria incubated at static head lowered delta GP by 1.1 kcal/mol and stimulated respiration by a factor of about 2.5 but caused negligible changes in delta GR1 + 2 and delta GR3. This observation demonstrates that the respiratory reactions from substrate to cytochrome c and from cytochrome c to oxygen both move away from thermodynamic equilibrium with delta GP during the transition from resting to active oxidative phosphorylation. The findings are discussed in terms of current schemes of chemiosmotic coupling.

Adenosine Triphosphate↗
PubMed1982

Separation of two thyrotropin binding components from porcine thyroid tissue by affinity chromatography: characterization of high and low affinity sites.

Two distinct thyrotropin (TSH) binding species have been separated from solubilized porcine thyroid membranes. Membranes was solubilized with 1% Triton X-100, and the supernatant was recovered by centrifugation at 105,000 X g. Scatchard analysis of thyrotropin binding to solubilized membranes (SM) yielded a nonlinear plot with Kd values for the high and low affinity components similar to those of intact membranes. Chromatography of the SM preparation on concanavalin A-Sepharose 4B resulted in the retention of 10-20% of the binding activity. Upon elution of the column, a peak of binding material (5-7% of total activity) was eluted at 0.3 M alpha-methyl-D-mannoside. This concanavalin A (Con A) bound fraction exhibited a linear Scatchard plot with a Kd value similar to that of the high affinity component of the SM. The protein fraction that did not bind to Con A (Con A unbound) also exhibited a linear Scatchard plot, but with affinity similar to that of the low affinity component of SM. Discontinuous sucrose density gradient ultracentrifugation revealed the presence of two major binding peaks in the solubilized membrane preparation. The slowly sedimenting peak corresponded to that seen in the Con A bound fraction, whereas the rapidly sedimenting peak corresponded to that of the Con A unbound fraction. Sepharose 6B chromatography indicated that in the case of the Con A unbound fraction, a single peak of specific binding activity was eluted in the void volume, and in the case of the Con A bound fraction, one major peak with an approximate Stokes radius of 67 A and several other minor peaks were eluted. These results demonstrate the physical separation of two distinct TSH binding species from thyroid membranes and provide further support for the model of multiple classes of binding sites.

Animals↗
PubMed1980

[Bioelectrode jellies for long-term monitoring (author's transl)].

The properties of 18 electrode jellies were investigated: electrical conductivity, ionic content, pH, skin irritation, and skin impedance reduction with different skin preparations, motion artifact, electrochemical noise with different electrode material, susceptibility to drying, ease of handling, and cost. There were significant differences of performance. Skin compatibility after 8 h ranged from "no irritation" to "severely irritating". A skin irritation most likely occurred with decornifying skin preparation. The specific skin impedance 24 min after the application ranged from 15 k omega . cm2 to 125 k omega . cm2. The motion artifacts differed up to 250%. All jellies produced negligible noise with Ag/AgCl and stainless steel electrodes. However, with gold and silver electrodes the noise amounted to 19 and 23 mu V. The NaCl content was correlated to the skin irritation and the impedance reduction. The impedance reduction correlated inversely to the skin compatibility and the motion artifact. Guidelines for the selection of an appropriate jelly in ergonomic long-term monitoring are given.

Cost-Benefit Analysis↗