Biomedical subjects

G Csúcs

Publications and source records attributed to G Csúcs.

7 recordsLinked to original sources

PubMed2004

Auto-reverse nuclear migration in bipolar mammalian cells on micropatterned surfaces.

A novel assay based on micropatterning and time-lapse microscopy has been developed for the study of nuclear migration dynamics in cultured mammalian cells. When cultured on 10-20-microm wide adhesive stripes, the motility of C6 glioma and primary mouse fibroblast cells is diminished. Nevertheless, nuclei perform an unexpected auto-reverse motion: when a migrating nucleus approaches the leading edge, it decelerates, changes the direction of motion, and accelerates to move toward the other end of the elongated cell. During this process, cells show signs of polarization closely following the direction of nuclear movement. The observed nuclear movement requires a functioning microtubular system, as revealed by experiments disrupting the main cytoskeletal components with specific drugs. On the basis of our results, we argue that auto-reverse nuclear migration is due to forces determined by the interplay of microtubule dynamics and the changing position of the microtubule organizing center as the nucleus reaches the leading edge. Our assay recapitulates specific features of nuclear migration (cell polarization, oscillatory nuclear movement), while it allows the systematic study of a large number of individual cells. In particular, our experiments yielded the first direct evidence of reversive nuclear motion in mammalian cells, induced by attachment constraints.

3T3 Cells↗

PubMed2003

Surface modification of PLGA microspheres.

Microspheres made of poly(lactic-co-glycolic acid) (PLGA) are biocompatible and biodegradable, rendering them a promising tool in the context of drug delivery. However, nonspecific adsorption of plasma proteins on PLGA micro- and nanospheres is a main limitation of drug targeting. Poly(L-lysine)-g-poly(ethylene glycol) (PLL-g-PEG), physisorbed on flat metal oxide surfaces, has previously been shown to suppress protein adsorption drastically. The goal of our work was to characterize the efficiency of the protein repellent character of PLL-g-PEG on PLGA microspheres and to show the feasibility of introducing functional groups on the PLGA microspheres via functionalized PLL-g-PEG. To quantify the adsorbed amount of protein, a semiquantitative method that uses confocal laser scanning microscopy (CLSM) was applied. The first part of the experiment confirms the feasibility of introducing specific functional groups on PLL-g-PEG-coated PLGA microspheres. In the second part of the experiment, PLL-g-PEG-coated PLGA microspheres show a drastic decrease of adsorbed proteins by two orders of magnitude in comparison to uncoated PLGA microspheres. Low protein-binding, functionalizable microspheres provide a fundamental basis for the design of drug delivery and biosensor systems.

Biosensing Techniques↗

PubMed2002

Optical grating coupler biosensors.

By incorporating a grating in a planar optical waveguide one creates a device with which the spectrum of guided lightmodes can he measured. When the surface of the waveguide is exposed to different solutions, the peaks in the spectrum shift due to molecular interactions with the surface. Optical waveguide lightmode spectroscopy (OWLS) is a highly sensitive technique that is capable of real-time monitoring of these interactions. Since this integrated optical method is based on the measurement of the polarizability density (i.e., refractive index) in the vicinity of the waveguide surface, radioactive, fluorescent or other kinds of labeling are not required. In addition, measurement of at least two guided modes enables the absolute mass of adsorbed molecules to be determined. In this article, the technique will be described in some detail, and applications from different areas will be discussed. Selected examples will be presented to demonstrate how monitoring the modification of different metal oxides with polymers and the response of the coated oxides to biofluids help in the design of novel biomaterials; how OWLS is useful for accurate bioaffinity sensing, which is a key issue in the development of new drugs; and how the quantitative study of protein-DNA/RNA and cell surface interactions can enhance the understanding of processes in molecular and cellular biology.

Adsorption↗

PubMed2001

The effect of UV irradiation on uracil thin layer measured by optical waveguide lightmode spectroscopy.

The polycrystalline uracil thin-layer dosimeter is a well-established method to monitor the biological effects of the environmental ultraviolet (UV) radiation. It is based on the optical density (OD) decrease of the uracil layer in the UV absorption band due to photodimerization of the crystal caused by UV irradiation. In the present study, we report measurements made with optical waveguide lightmode spectroscopy (OWLS) to characterize the changes in the optogeometrical parameters of the uracil layer caused by an artificial UV source. It is shown that UV irradiation causes a decrease in the refractive index and an increase of the optical anisotropy. The determined kinetic parameters of the UV dose-sensor response curves correlate well with results of OD measurements, but the sensitivity of OWLS is about ten times higher. The results show that OWLS is capable of analyzing the UV response of the uracil layer and opens the way for dosimetrical applications.

Biosensing Techniques↗

PubMed1998

Solubilization of planar bilayers with detergent.

The interaction of the nonionic detergent Triton X-100 with supported phosphatidylcholine planar lipid bilayers has been investigated by optically monitoring changes in the bilayer, using the technique of optical waveguide lightmode spectroscopy (OWLS). This technique has several advantages over the methods applied to the problem hitherto, including: high sensitivity; measurement in situ with good time resolution; the fact that the free detergent concentration is well-defined, and the lipid concentration in solution is zero; ease of studying the reversibility of the interaction; and the readiness with which absolute rather than effective amounts of detergent incorporated into the lipid can be determined. The main finding is that as the free Triton concentration increases, the detergent is first incorporated reversibly into the bilayer, then partly but never completely removes lipid, and finally (at or above the cmc) completely solubilizes the bilayer. The behaviour of the lanar supported lipid bilayers is thus similar to that previously reported for lipid vesicles.

Detergents↗

PubMed1998

Interaction of phospholipid vesicles with smooth metal-oxide surfaces.

The interaction of phospholipid vesicles with planar metal oxide supports has been previously reported as a means of preparing supported lipid bilayers, which are useful models of biological membranes. Nevertheless, extant evidence that bilayers are actually formed is rather circumstantial, and the necessary and sufficient conditions for their formation have never been delineated. Here, we tackle this problem by using smooth planar optical waveguides as the support. Analysis of the lightmode spectra of the waveguides, measured in situ during the deposition process, yields the mass of lipid deposited at the solid/liquid interface. By comparing the optogeometric parameters of the structures assembled from the vesicles with those of a lipid bilayer of known structure assembled using the Langmuir-Blodgett technique, we show that in many cases the vesicles remain intact and form a supported layer of vesicles rather than a bilayer, and often mixed structures (intact vesicles embedded in a bilayer partially covering the surface) occur. Careful analysis of the lipid deposition kinetics corroborates this result. We have also found that divalent cations dramatically promote attachment of mixed phosphatidylcholine/phosphatidylglycerol vesicles to form supported vesicle layers, and bilayer formation from pure phosphatidylcholine vesicles.

Adsorption↗

PubMed1996

An increased intensity of N-methyl-D-aspartate (NMDA) but not non-NMDA receptor activation may be responsible for the enhancement of excitatory processes in the neocortex of two-week-old rats: a brain slices study.

During the brain maturation a critical period is detectable when the sensitivity of the neocortex is high. Enhanced excitatory activity is characteristic at that time while the inhibitory processes are underdeveloped. The goal of this study was to determine the effectiveness of different types of excitatory amino acid antagonists reducing the electrically evoked excitatory synaptic responses of the somatosensory cortex. Effects of the specific competitive N-methyl-D-aspartate (NMDA) antagonist 4-amino-phosphono-valerate (APV), and the specific non-competitive, non-NMDA antagonist 1-(4-aminophenyl)-3-methylcarbamoyl-7,8-methylenedioxy-5H-2,3-benz odiazepine (GYKI 53655) were analysed on neocortex slices prepared from 2-week-old and adult rats. APV caused a partial inhibition of the electrically evoked response more effectively in young animals than in adults, while the effective IC50 values were similar. In contrast, the non-NMDA antagonist had a similar effect on the slices of both age-groups.

Action Potentials↗