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Biomedical subjects

M Kveder

Publications and source records attributed to M Kveder.

15 recordsLinked to original sources
PubMed2001

The action of mercury on cell membranes.

The action of mercuric chloride and methyl mercuric chloride on the membrane lateral domain organization of bovine, equine, and canine erythrocytes was studied. Electron paramagnetic resonance (EPR) spectra of spin-labeled erythrocytes were analyzed with respect to their lateral domain structure. Continuous alteration of the membrane domain populations revealed that mercuric compounds affect the membrane via the evolution of toxic events in the cells.

Animals↗
PubMed2000

Spectroscopic evidence of beta-turn in N-glycated peptidomimetics related to leucine-enkephalin.

The conformational differences caused by N-glycation of the amide bond in endogenous opioid pentapeptide leucine-enkephalin (Tyr-Gly-Gly-Phe-Leu) have been explored in solution using FTIR spectroscopy, NMR and molecular modelling. The compounds studied include protected and unprotected enkephalin analogues N-alkylated at the second (Gly2) amino acid residue with a 6-deoxy-D-galactose moiety (1-3). Comparison of the amide I component bands in the FTIR spectra, measured in trifluoroethanol (TFE), CHCl3 and DMSO, revealed significant differences in the intensity as well as shifts in component band frequencies for glycopeptides 1-3. We found that only the FTIR spectrum of the fully protected compound 1 indicated the presence of a higher population of beta-turns, while the spectra of the partially protected and unprotected glycopeptides 2 and 3 reflected the dominance of unordered or open structures, with some low population of turns. The observed NOE connectivities in CDCl3 for both isomers of the fully protected compound 1, the all-trans one and another with Tyr1-Gly2 peptide bond in cis conformation, indicate the presence of a beta-like turn conformation. Molecular dynamics simulations of the glycopeptide 1 obtained by unconstrained energy minimization of trans- and cis-1 shows that one of trans form conformations is consistent with beta-turn whereas cis isomer has revealed less-compact turn.

Enkephalins↗
PubMed2000

Structural aspects of thiol-specific spin labeling of human plasma low density lipoprotein.

A novel thiol-specific spin labeling procedure for the protein component (apoprotein B, apoB) of low density lipoproteins (LDLs) is presented. A methanethiosulfonate spin label was used to probe the free cysteine residues of apoB with electron paramagnetic resonance (EPR) spectroscopy. The results indicated that the spin labeled sites are predominantly buried in the LDL particle in two distinct environments that differ in their mobility restrictions. The suitability of thiol-specific labeling for the study of the stability and conformation of apoB was demonstrated in experiments with denaturing agents. The results presented in this work offer a new approach for the matching of EPR data with the primary structure of apoB.

Apolipoproteins B↗
PubMed2000

The EPR study of LDL perturbed by alcohols with different molecular architecture.

In this work, the interaction of different isomers of lower aliphatic alcohols with LDL representing a complex macromolecular assembly is investigated in vitro. Emphasis is given to the comparison of the impact of molecular architecture of methanol, ethanol, propanol (n-, iso-) and butanol (n-, iso-, sec-, tert-) in perturbing the lipid-protein assembly. The geometrical characteristics as well as the lipophilicity of the respective alcohol are considered. The EPR method combined with the spin labeling of both the apoB and the lipid monolayer allowed parallel detection of changes provoked in both phases. In addition to the change in protein environment, the spectral decomposition of the experimental data revealed a decrease in lipid ordering with the increasing concentration of the alcohols. This phenomenon for aliphatic alcohols is linearly correlated with the equal volume occupation (EVO) of alcohol in LDL. The results support the molecular mechanism of alcohol action through its interference with the lipid-protein interactions in LDL, which could be applicable to the molecular mechanism of alcohol interaction with integral membrane proteins.

1-Propanol↗
PubMed1997

The interaction of lower alcohols with apoB in spin labeled human plasma low density lipoproteins (LDL).

In this study the interaction of alcohol with the macromolecular lipid-protein assembly represented by human plasma low density lipoproteins (LDL) was investigated. The spin label which covalently binds to the side chain amino group of lysines as well as terminal amino groups was attached to the spin labeled apoprotein (apoB) of native LDL in order to observe the protein component in the electron spin resonance (ESR) spectrum. The interaction of different lower alcohols (methanol, ethanol, propanol and butanol) with the spin labeled LDL was studied for two alcohol concentrations (0.3 and 3.0 M). The ESR spectra indicate a decrease of the hyperfine splitting and narrowing of the linewidth upon the action of alcohol that leads to the conclusion that alcohol provokes a change in the apoB conformation. These findings are explained by following the arguments of the phospholipid mediated mechanism of alcohol action, through the modulation of the lipid packing free volume which results in the protein conformational change.

Alcohols↗
PubMed1997

Nitroxide reduction with ascorbic acid in spin labeled human plasma LDL and VLDL.

The LDL and VLDL were spin labeled with Tempo which partitions both in the aqueous and lipid phase. The ESR spectra were measured in the equilibrium state as well as during the reduction of the spin label with ascorbic acid. The kinetics of the concentration decay curves was parametrized with two exponentials. The theoretical simulation of the experimental spectra revealed a drastic linewidth narrowing in the VLDL samples exposed to the ascorbic acid. Since the transport properties of the specific monolayer are reflected in the observed reaction rates, the analysis of the fatty acid composition of phospholipids, triglycerides and cholesterol esters in LDL and VLDL was performed. It is concluded that different lipid packing at the surface of LDL and VLDL might be the consequence of different intermolecular forces between phospholipids and cholesterol. This finding was connected to the experimentally detected different reaction kinetics in LDL and VLDL as well as their different susceptibility to the ESR linebroadening effects during the nonequilibrium conditions of the spin label reduction with ascorbic acid.

Ascorbic Acid↗
PubMed1995

Three-dimensional structure of bovine heart fatty-acid-binding protein with bound palmitic acid, determined by multidimensional NMR spectroscopy.

The three-dimensional structure of the holo form of recombinant cellular bovine heart fatty-acid-binding protein (H-FABPc), a polypeptide of 133 amino acid residues with a molecular mass of 15 kDa, has been determined by multidimensional homonuclear and heteronuclear NMR spectroscopy applied to uniformly 15N-labeled and unlabeled protein. A nearly complete set of 1H and 15N chemical shift assignments was obtained. A total of 2329 intramolecular distance constraints and 42 side-chain chi 1 dihedral-angle constraints were derived from cross-relaxation and J coupling information. 3D nuclear Overhauser enhancement and exchange spectroscopy combined with heteronuclear multiple-quantum coherence (NOESY-HMQC) experiments, performed on a sample of uniformly 13C-labeled palmitic acid bound to unlabeled cellular heart fatty-acid-binding protein revealed 10 intermolecular contacts that determine the orientation of the bound fatty acid. An ensemble of protein conformations was calculated with the distance-geometry algorithm for NMR applications (DIANA) using the redundant dihedral-angle constraint (REDAC) strategy. After docking the fatty acid into the protein, the protein-ligand arrangement was subject to distance-restrained energy minimization. The overall conformation of the protein is a beta-barrel consisting of 10 antiparallel beta-strands which form two nearly orthogonal beta-sheets of five strands each. Two short helices form a helix-turn-helix motif in the N-terminal region of the polypeptide chain. The palmitic acid is bound within the protein in a U-shaped conformation close to the two helices. The obtained solution structure of the protein is consistent with a number of fatty-acid-binding-protein crystal structures.

Amino Acid Sequence↗
PubMed1994

The ESR characterization of molecular mobility in the lipid surface layer of human serum lipoproteins.

Three different nitroxides were used to probe either the head group (Tempil stearate) or acyl chain region (Spin labeled cholestane (ChSl) and methyl ester of 5 doxyl palmitate (MeFASL(10,3))) of human plasma low density lipoproteins (LDL) and very low density lipoproteins (VLDL). The ESR data were compared with the simulated spectra which assume rapid anisotropic motion of nitroxide. The results indicate that in the head group region of both LDL and VLDL only the slowing down of the rotational motion occurred when temperature was lowered and the whole region showed up as a unique compartment. On the other hand, the acyl chain region, probed with MeFASL(10,3), behaved as one compartment at physiological temperatures, while at lower temperatures coexistence of fluid and immobilized components were observed. The ESR spectra of lipoproteins labeled with Cholestane showed even higher sensitivity to the mobility constraints. Here, the LDL spectra revealed a drastic immobilization of ChSl axial rotation already at physiological temperatures. The results of these experiments were discussed in terms of core phase transition and/or lipid-protein interactions.

Electron Spin Resonance Spectroscopy↗
PubMed1988

Non-Brownian water self-diffusion in lung tissue.

Pulsed field gradient NMR measurements of rat lung tissue have shown that the apparent water self-diffusion coefficient is nearly an order of magnitude smaller than that of free water and moreover is not constant but varies as the inverse square root of the diffusion time. The mean square displacement of water molecules is similarly proportional to the square root of the diffusion time. Possible origins of this non-Brownian behavior are discussed.

Animals↗
PubMed1988

Water proton NMR relaxation mechanisms in lung tissue.

The NMR relaxation times T'2, T2, and T1 were measured in isolated rat lungs as functions of external magnetic field B0, temperature, and lung inflation. The observed linear dependence on B0 of the tissue-induced free induction decay rate (T'2)-1 provides independent confirmation of the air/water interface model of the lung. Furthermore, measurements of the Larmor frequency dependence of T1 are consistent with a spin-lattice relaxation rate of the form 1/T1 = A omega -1/2 + B as expected for the case in which the relaxation arises from water-biopolymer cross-relaxation, which should be proportional to the surface area of the lung. This prediction was verified by observations of an approximately linear dependence of 1/T1 on transpulmonary pressure and thus on the lung surface area.

Animals↗
PubMed1988

Spin probe reduction in cells and tissues.

The reduction rates of different nitroxides in rat lung tissue were measured by electron paramagnetic resonance. We confirmed that the reduction rate of nitroxide spin probe molecules is coupled with their structure and transport characteristics. Oxygen was found to slow down the reduction rate of nitroxides in rat lung tissues. On the basis of these findings it can be concluded that most nitroxide properties described for homogeneous systems--cells and tissue homogenates--are also valid for heterogeneous systems, which is important for the application of nitroxides as metabolically active NMR contrast agents.

Aerobiosis↗
PubMed1987

Theoretical aspects of multiple deconvolution analysis for quantification of left to right cardiac shunts.

A new method for quantification of left to right cardiac shunts by Bourguignon et al based on multiple deconvolution analysis is critically analysed within the framework of a simple mathematical model. Underlying assumptions are explicitly stated and their validity discussed. It turns out that some reinterpretation of the method is necessary. Using the same ideas as in multiple deconvolution analysis, a new relation for the pulmonary to systemic flow ratio is proposed on a theoretical basis. This technique may be useful when diagnosing left to right cardiac shunts with radiocardiographic methods.

Coronary Circulation↗
PubMed1985

A mathematical model for the quantitative study of left to right cardiac shunt.

Non-invasive radioisotope cardiographic techniques have become a useful tool for studying the anatomy and function of the heart. The mathematical model described justifies the phenomenological analysis of pulmonary time-activity histograms and was invented for quantitative study of left to right cardiac shunt. The model also gives a theoretical insight into such an empirically proposed diagnostic method and represents an adequate framework for the understanding of other possible approaches to the problem of left to right shunt.

Blood Circulation↗
PubMed1983

A study of explicit greytone perception on TV video monitors.

46 persons have been tested in their ability to resolve grey tones in quantitatively callibrated transparencies. The transparencies were designed as phantoms matched in lightness and dynamics to TV monitors. The results show that on average not more than 9 grey tones can be counted in such phantoms and that dark gaps between the fields decrease the confidence of viewers.

Data Display↗