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Biomedical subjects

P Gróf

Publications and source records attributed to P Gróf.

At least 19 recordsLinked to original sources

Studies on molecular interactions between nalidixic acid and liposomes.

The interaction between nalidixic acid sodium salt (NANa) and liposomes prepared from alpha-L-dipalmitoyl-phosphatidylcholine (DPPC) or from its binary mixture with dioleoyl-phosphatidylcholine (DOPC) was studied with differential scanning calorimetry (DSC) and electron paramagnetic resonance (EPR) spectroscopy. We evaluated the role of broadband ultraviolet-B (UV-B) irradiation on the molecular interactions between the lipids and the NANa, and determined the decay-kinetics of the incorporated spin labeled fatty-acid free radicals. Multilamellar and unilamellar vesicles were prepared by sonication and extrusion. The entrapment efficiencies were determined spectrophotometrically. The size-distribution of the liposomes and its change in time was checked by dynamic light scattering (DLS). Our results indicate that NANa mainly interacts with lipid head groups. However, its effect and presumably the formation of the free radicals, induced by broadband ultraviolet-B, is not localized only to the head group region of the lipid molecules. Depending on DOPC content, interaction between the NANa and the lipids modifies the phase-transition parameters of the liposome dispersions.

1,2-Dipalmitoylphosphatidylcholine↗

Biological UV dosimeters in simulated space conditions.

Polycrystalline uracil thin layers participate in the phage and uracil response (PUR) experiment, assigned to the biological dosimetry of the extraterrestrial solar radiation on the International Space Station (ISS). In ground based experiments (experiment verification tests), the following space parameters were simulated and studied: temperature, vacuum and short wavelength UV (UV-C, down to 200 nm) radiation. The closed uracil samples proved to be vacuum-tight for 7 days. In the tested temperature range (from -20 to +40 degrees C) the uracil samples are stable. The kinetic of dimer formation (dimerization) and reversion (monomerization) of uracil dimers due to short wavelength UV radiation was detected, the monomerization efficiency of the polychromatic deuterium lamp is higher than that of the germicidal lamp. A mathematical model describing the kinetic of monomerization-dimerization was constructed. Under the influence of UV radiation the dimerization-monomerization reactions occur simultaneously, thus the additivity law of the effect of the various wavelengths is not applicable.

Dimerization↗

Biological UV dosimeters in quality control of tanning tubes.

Although according to the International Radiological Protection Association-International Non-Ionizing Radiation Committee recommendation (1991) the use of sunbeds for cosmetic purposes is not recommended, tanning devices are used widely. Ten different types of commercially available sunbed tubes have been studied using a uracil biological UV dosimeter, and three of them were analyzed in detail. Dimerization effectiveness of the tubes was measured directly, whereas efficiency of erythema induction was calculated weighting the emission spectra by the Commission Internationale de l'Eclairage erythema action spectrum. The data obtained demonstrate that quality control of sunbed tubes has to include not only the determination of the UV doses administered but also the assessment of the health risk due to the UVB and UVA components of the lamp. A method of quality control using the uracil biological dosimeter was elaborated, and the estimation of the "acceptable" exposure time was checked/controlled on 15 volunteers by assessing individually the erythema induction threshold. A correct classification of the sunbed tubes is proposed by characterizing the erythema induction versus DNA-damaging effectiveness of tubes.

Heliotherapy↗

[Preparations of liposomal morphine. Investigation of the interaction of morphine and its derivatives with the dipalmitoyl phosphatidylcholine membrane].

Liposomes composed of the lipids of biological membranes are suitable for drug delivery. To reach a better therapeutical effect it is important to know the properties of interactions between the drug and lipid molecules. From dipalmitoyl phosphatidylcholine (DPPC) using ultrasound small liposomes containing morphine were prepared. The amount of entrapped morphine was determined with spectrophotometry and luminescence spectroscopy. The interaction between the molecules of morphine and its derivates (codeine, N-methyl-morphine, N-methyl-codeine) and the DPPC lipid was studied with differential scanning calorimetry (DSC) and electron spin resonance (ESR) methods. Our studies indicated that the molecules of morphine and its derivates principally interact with the environment of DPPC lipid head groups. Due to the interaction the mobility of head groups decreases especially in case of codeine and N-methylcodeine.

1,2-Dipalmitoylphosphatidylcholine↗

Oxidation of guanine in cellular DNA by solar UV radiation: biological role.

The formation of cyclobutane pyrimidine dimers (CPD) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodGuo) was investigated in Chinese hamster ovary cells upon exposure to either UVC, UVB, UVA or simulated sunlight (SSL). Two cell lines were used, namely AT3-2 and UVL9, the latter being deficient in nucleotide excision repair and consequently UV sensitive. For all types of radiation, including UVA, CPD were found to be the predominant lesions quantitatively. At the biologically relevant doses used, UVC, UVB and SSL irradiation yielded 8-oxodGuo at a rather low level, whereas UVA radiation produced relatively higher amounts. The formation of CPD was 10(2) and 10(5) more effective upon UVC than UVB and UVA exposure. These yields of formation followed DNA absorption, even in the UVA range. The calculated relative spectral effectiveness in the production of the two lesions showed that efficient induction of 8-oxodGuo upon UVA irradiation was shifted toward longer wavelengths, in comparison with those for CPD formation, in agreement with a photosensitization mechanism. In addition, after exposure to SSL, about 19% and 20% of 8-oxodGuo were produced between 290-320 nm and 320-340 nm, respectively, whereas CPD were essentially (90%) induced in the UVB region. However, the ratio of CPD to 8-oxodGuo greatly differed from one source of light to the other: it was over 100 for UVB but only a few units for UVA source. The extent of 8-oxodGuo and CPD was also compared to the lethality for the different types of radiation. The involvement of 8-oxodGuo in cell killing by solar UV radiation was clearly ruled out. In addition, our previously reported mutation spectra demonstrated that the contribution of 8-oxodGuo in the overall solar UV mutagenic process is very minor.

Animals↗

Changes of phage T7 nucleoprotein structure at low ionic strength. A Raman spectroscopic study.

To detect changes in DNA and/or protein structures of phage T7 under different ionic strength, Raman spectra of phage T7 have been recorded in solutions of three different NaCl + Tris concentrations. Iterative Jansson-Van Cittert deconvolution, as well as decomposition methods have been used to quantify changes in DNA structure. Significant modifications in ratios of contributions from 675 and 685 per cm vibrations, as well as in the DNA backbone vibrations, characteristic for B-DNA, near 835 per cm frequency have been found. Changes of the base electronic structure were identified in the interval between 1280 and 1400 cm(-1). Estimation of the overall protein structure suggests predominant beta-sheet content.

Bacteriophage T7↗

Use of uracil thin layer for measuring biologically effective UV dose.

Dimerization of uracil monomers in a polycrystalline state by UV radiation changes the absorption characteristics of a thin layer of the material. The change in optical density, measured by spectrophotometry in the 250-400 nm range, as a function of the exposure time is evaluated in terms of the biologically effective UV dose. A statistical evaluation of a great number of uracil dosimeters irradiated with a TL01 lamp from Philips establishes the possibility of evaluating the biologically effective UV dose using a uracil dosimeter. Nonlinear regression procedures were introduced to correct the absorption spectra for contributions due to light scattering and to determine the optical density values required to calculate the UV dose expressed in HU units. Comparison of cumulative daily doses and long-term monitoring measured by the uracil thin-layer dosimeter and a phage T7 dosimeter are given, which allow the determination of conversion factors between various biological dosimeters under different irradiation conditions.

Dose-Response Relationship, Radiation↗

Raman spectroscopic study of conjugates of butyrylcholinesterase with organophosphates.

Raman spectra of human butyrylcholinesterase (BuChE; E.C. 3.1.1.8) were analyzed in the native state and after conjugation with organophosphates (soman, DFP and paraoxon). The secondary structure of the native BuChE in Tris-HCl buffer (pH 7.5), determined from analysis of the amide I polypeptide vibration band, indicates 47% alpha-helices, 26% beta-sheets, 16% turns and 12% undefined structure. We obtained the same values for paraoxon-phosphorylated BuChE, but 39% helical structure, 31% beta-sheets, 17% turns and 13% undefined structure for 'aged' DFP-BuChE conjugates and 36% helical structure, 34% beta-sheets, 20% turns and 10% undefined structure for 'aged' soman-BuChE conjugates. The approximately 10% decrease of alpha-helical structure observed upon phosphorylation by DFP and phosphonylation by soman, probably corresponds to the 'aging' process, which does not take place in the case of paraoxon. Considerable differences have been observed between native, paraoxon inhibited and 'aged' BuChE in aromatic ring vibrations, suggesting that the dealkylation of organophosphate conjugates modifies the environment or the interactions of aromatic amino-acid residues. In the aliphatic side chains an increase of the number of gauche configurations has been observed in 'aged' DFP-BuChE and soman-BuChE.

Butyrylcholinesterase↗

Genotoxic effectivity--comparison of 36 nitrated furan and arenofuran derivatives on a quantitative scale. Statistical comparison of T7 and other short-term tests.

Thirty-six nitrated furan and arenofuran derivatives were measured and quantitatively characterized by the T7 inactivation test. A wide range of substances previously studied allowed us to compare the collected quantitative data with those obtained by other workers using different short-term tests. Based on comparative statistical evaluation of these data a borderline was determined for the genotoxic effect: compounds having in our short-term test mutagenicity index (MI) values smaller than 8.0 are positive while the higher values represent negative genotoxicity. Classification of 36 nitrofuran/nitroarenofuran derivatives is given both on the basis of the quantitative genotoxicity scale and in terms of +/- on the qualitative scale. All but one compound were found to be genotoxic and the genotoxic activities of these compounds were compared with the results of other carcinogenicity or mutagenicity tests.

Carcinogenicity Tests↗

Raman spectroscopic study on the conformation of 11 S form acetylcholinesterase from Torpedo californica.

Vibrational Raman spectroscopy has been used to study the conformation of the 11 S form of acetylcholinesterase from Torpedo californica. Secondary structure analysis by the method of Williams [(1983) J. Mol. Biol. 166, 581-603] shows 49% alpha-helical structure, 23% beta-sheets, 11% turns and 15% undefined structure. Secondary structure estimates obtained for this enzyme by Raman spectroscopy and circular dichroism have been analyzed.

Acetylcholinesterase↗

The role of DTNB light chain in the contractile properties of skeletal muscle.

The mechanical and molecular dynamical properties of the glycerinated muscle fibres from m. psoas of rabbit were investigated after treatment with 5,5'-dithiobis/2-nitrobenzoate (DTNB). After DTNB treatment the fibre bundles shortened and exerted force in ATP-relaxing solution. The rate of the ATP hydrolysis in relaxation-inducing medium increased significantly after DTNB treatment, and simultaneously the amount of the LC2(DTNB) light chain decreased to 40 per cent of the original value measured by gel electrophoresis. The electron paramagnetic resonance spectra of muscle fibres labelled with maleimide spin label after DTNB treatment exhibited high degree of order of label in rigor, but this order decreased after shortening. The experiments support the assumption that, in vertebrate skeletal muscles, besides the troponin-tropomyosin regulation system which developed later in evolution, the LC2 light chain should maintain some regulatory properties.

Adenosine Triphosphatases↗

Effect of DTNB-treatment on Ca++-sensitivity of superprecipitation and ATPase activity of actomyosin.

A light chain of 18 000 daltons of native actomyosin isolated from rabbit skeletal muscle was removed by DTNB-treatment. Investigated were the differences in superprecipitation and ATPase activity of LC2-deficient and control actomyosin. The superprecipitation of control actomyosin develops in two phases, with high amplitude and kinetics dependent on the Ca++ concentration. On the other hand, superprecipitation of treated actomyosin develops in a single phase, with low amplitude and a kinetics independent of the Ca++ concentration. The partial lack of LC2 leads to the loss of Ca++-sensitivity of ATPase activity. On the basis of the results, LC2 has been assumed to fix that conformation state of myosin heads which is required for the functioning of the troponin system on the one hand, and a cooperation between myosin heads on the other. The regulatory function of LC2 is manifest in controlling the actin-myosin-ATP interaction, at the activated state of the troponin system, in an extent depending on the actual Ca++ concentration.

Actomyosin↗

[Formation of secretory immunoglobin by the hepatic clearance mechanism].

In patients suffering predominantly from several types of dermatosis as well as from cholecystitis and cholelithiasis relationships to the formation of secretory immunoglobulin by the hepatic clearance mechanism have been investigated. In the duodenal content and in operatively yielded gall-bladder bile, immunoglobulins, complement C3, and albumin were detected. The amount of immunoglobulins and of albumin in the duodenal content was substantially increased if infectious germs could be detected in the stomach content.

Bile↗

Molecular motions in myosin head regions of dog cardiac muscle fibers.

Rotational dynamics of myosin heads in glycerinated cardiac muscle fibres were studied by using the spin-labelling technique in the conventional and saturation transfer electron paramagnetic resonance time domains. The maleimide spin labels which were attached to the fast-reacting thiols (SH1 or SH2, respectively) of the myosin moiety, exhibited orientational order with respect to the fibre axis in rigor state of the muscle fibres; the higher degree of order of the paramagnetic probe molecules was obtained when the labels were attached to the SH2 thiol sites. The degree of order of spin labels is reduced during muscle activity. In rigor the rotational correlation time of the labels bound to the SH2 sites was in the submillisecond time range (tau 2 approximately 30 microseconds), whereas tau 2 was equal to about 0.1 microseconds when the labels occupied the SH1 sites. The comparison of electron paramagnetic resonance spectra on cardiac and skeletal muscle indicates differences in the mobility of spin labels around the fast reacting thiols of the two myosins and also in the subpopulations of the labelled sites. On the basis of the rotational correlation times it can be suggested that a fraction of myosin heads might be free and/or the attached heads have some residual mobility or large degree of flexibility in rigor of the cardiac muscle at complete filament overlap.

Animals↗

The effect of anaesthetics on protein conformation in membranes as studied by the spin-labelling technique.

The effect of general and local anaesthetics was studied on the membrane components (proteins and lipids) of frog sciatic nerve using the spin-labelling technique. A rapid and effective method was developed to incorporate fatty acid labels into the lipid regions and to attach the maleimide spin labels to the thiol sites of the proteins of the membrane. On the basis of the rotational correlation time of the attached maleimide spin labels, at least three different protein domains were identified. One part of the labels has a preferred orientation with respect to the axis of the nerve fibre. The effect of halothane and of local anaesthetics such as lidocaine and tetracaine, which influence primarily the lipid regions of the membrane, is efficiently transferred to the spin-labelled membrane proteins via strong lipid-protein interaction. The results support the concept that the architecture and the physiological activity of the membrane-bound proteins are sensitive to changes in the physical state of membrane lipids.

Action Potentials↗