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Biomedical subjects

M Langner

Publications and source records attributed to M Langner.

At least 19 recordsLinked to original sources

Abrupt transition in quasiparticle dynamics at optimal doping in a cuprate superconductor system.

We report time-resolved measurements of the photoinduced change in reflectivity, DeltaR, in the Bi2Sr2Ca(1-y)Dy(y)Cu2O8+delta (BSCCO) system of cuprate superconductors as a function of hole concentration. We find that the kinetics of quasiparticle decay and the sign of DeltaR both change abruptly where the superconducting transition temperature T(c) is maximal. These coincident changes suggest that a sharp transition in quasiparticle dynamics takes place precisely at optimal doping in the BSCCO system.

Journal Article↗

Combined effect of surface electrostatic charge and poly(ethyl glycol) on the association of liposomes with colon carcinoma cells.

Particulate drug formulations are considered to be a means that may improve the pharmacokinetics and biodistribution of active compounds. By using them, drug distribution is determined solely by the properties of the carrier. The surface properties of such supramolecular aggregates determine how they will interact with various biological structures. Among others, surface electrostatic charge and surface grafted polymers are considered to be among the major factors affecting its interaction with proteins and cells. In this article, we present experimental evidence that properly selected surface electrostatic charge and grafted polymers can alter the association of liposomes with colon cancer cells. The dependence of the adsorption of liposomes onto the cell surface on the quantity and length of surface grafted polymers for a certain surface charge density exhibits a distinct maximum. For example, when liposomes were formed with 20 mol% of DOTAP, PE-PEG350 increased liposome adsorption by up to 6 mol%. This adsorption maximum depends on both polymer length and charge type. Results presented in this article show that the interaction of liposomes with colon cancer cells can be tuned by a proper combination of liposome surface electrostatics and surface grafted polymers.

Cell Line, Tumor↗

ESR and monolayer study of the localization of coenzyme Q10 in artificial membranes.

The data obtained from the ESR experiments show a complex, depth dependent effect of CoQ10 on the lipid molecules mobility in the bilayer. These effects depend both on its concentration and the temperature. CoQ10 disturbs not only the hydrophobic core of the membrane but also the region close to the hydrophilic headgroups of phospholipids. Both these effects could be explained by the fact that the high hydrophobicity of CoQ10 causes the molecules to position itself in the interior of the bilayer, but at the same time its water seeking headgroup is located close to the region of the polar headgrops of membrane lipids. The presence of CoQ10 in the hydrophobic core has further implications on the properties of membrane intrinsic domain. Results of monolayer experiments indicate that CoQ10 may form aggregates when mixed with PC molecules in the lipid hydrocarbon chain-length dependent manner. CoQ10 is not fully miscible with DMPC or DPPC but it is well miscible with the long-chain DSPC molecules. Our suggestion is that CoQ10 when present in long-chain phospholipid bilayer, interacts with saturated fatty acyl-chains and adapt the structure which allows such interactions: either parallel to the saturated acyl chains or "pseudo-ring" conformation resembling sterol structure.

Coenzymes↗

Hereditary spherocytosis: identification of several HS families with ankyrin and band 3 deficiency in a population of southwestern Poland.

Red blood cells of 17 patients out of seven families diagnosed with HS from the southwest of Poland were studied. In six families a deficiency of ankyrin was detected, and in one family a band 3 (anion-exchanger protein) deficiency was detected. Patients from six families with the ankyrin deficiency had a 19-51% decrease in ankyrin 2.1, while the family with the band 3 deficiency showed a 33% decrease in this protein content. All changes were statistically significant, as analysed by the Student t test (P<0.05). Analysis of haemolysis kinetics gives a reliable indication of altered osmotic properties of the spherocytic cells.

Adolescent↗

The effect of steric constraints on the adsorption of phenyltin onto the dipalmitoylphosphatidylcholine bilayer.

In this paper, we present studies concerning phenyltin adsorption onto the dipalmitoylphosphatidylcholine bilayer. Phenyltin compounds are known to be biologically active, and their molecular geometry makes it possible to study the effect of steric constraints on their ability to penetrate the model lipid membrane. Using a fluorescence probe as a reporter of the amount of adsorbed compound, we evaluated their affinity to the membrane as a function of the membrane state. The amount of the adsorbed compound was found to depend on the adsorbing molecule's geometry and lipid bilayer organization. The fluorescence measurements were supported by the density functional theory (DFT) method of quantum mechanical computations. The penetrant location was correlated with the possible relative positions of its polar and hydrophobic moieties to determine if it could adopt structural requirements of the local membrane environment. Molecules were deformed by a model force, mimicking interactions within the membrane interfacial region. Computations show that the diphenyltin molecule can be deformed to such an extent that it can adopt an amphiphilic conformation. Triphenyltin is different, as its bending requires more energy. Born repulsion energies from hydrophobic fluid into water for phenyltins were also computed in an isodensity-polarized continua model of DFT computation. Our results indicate that the phenyltin compounds incorporate into the interface of the lipid membrane, although diphenyltin integrates more deeply than triphenyltin, which locates on the double layer's surface, and this is due to the fact that the main role is played by steric and not electrostatic interactions.

1,2-Dipalmitoylphosphatidylcholine↗

Effect of free fatty acids on the permeability of 1,2-dimyristoyl-sn-glycero-3-phosphocholine bilayer at the main phase transition.

We measured the influence of saturated and unsaturated free fatty acids on the permeability and partition of ions into 1, 2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) bilayers. The bilayer permeability was measured using the depletion of N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1, 2-dihexadecanoyl-sn-glycero-3-phosphatidylethanolamine (N-NBD-PE) fluorescence as a result of its reduction by dithionite. We observed a distinct increase of dithionite permeability at the main gel-fluid phase transition of DMPC. When vesicles were formed from a mixture of DMPC and oleic acid, the membrane permeability at the phase transition was reduced drastically. Stearic acid and methyl ester of oleic acid have little effect. Similar results in the quenching of pyrene-PC in DMPC vesicles by iodide were obtained. Again, the increase of iodide partition into the lipid phase at the main phase transition of DMPC was abolished by the addition of unsaturated free fatty acids. Free fatty acids, in concentrations up to 5 mol%, do not abolish DMPC phase transition when measured by differential scanning calorimetry. It seems that unsaturated, but not saturated, free fatty acids reduce the lipid bilayer permeability to dithionite and iodide ions at the main phase transition of DMPC, without altering the thermodynamic properties of the bilayer.

Dimyristoylphosphatidylcholine↗

Merocyanine 540 as a fluorescence indicator for molecular packing stress at the onset of lamellar-hexagonal transition of phosphatidylethanolamine bilayers.

The fluorescence of Merocyanine 540 (MC 540) is sensitive to the molecular packing of membrane lipids. Therefore, the fluorescence of MC 540 is expected to be sensitive to the curvature-related packing stress at the onset of the lamellar-hexagonal phase transition. We measured the fluorescence intensity of MC 540 when the temperatures of lipid bilayers approached their lamellar-hexagonal phase transitions. The fluorescence of MC 540 in the presence of egg and dioleoylphosphatidylethanolamine bilayers increased at the respective lamellar-hexagonal phase transitions of these lipids. Furthermore, increases in fluorescence intensity were also observed at temperatures just below their phase transitions. The enhanced fluorescence was not due to the specific interaction of the dye with the ethanolamine headgroup, because no such increase was observed when the probe was exposed to phosphatidylethanolamines which do not form hexagonal phase within the range of applied temperature. In addition, when the temperature of the lamellar-hexagonal phase transition was shifted, by the addition of a small amount of phosphatidylcholine, the dependence of the fluorescence intensity on temperature was modified accordingly. We postulate that the change of MC 540 fluorescence intensity at temperatures approaching the lamellar-hexagonal phase transition reflects changes in the partition of MC 540 into the fluid lipid phase. The change in partition is influenced by the curvature stress in bilayers at temperatures just below the lamellar-hexagonal phase transition.

Fluorescent Dyes↗

The electrostatics of lipid surfaces.

Charged lipids constitute a substantial fraction of all membrane lipids. Their charges vary in quantity and distribution within their headgroup regions. In long range interactions, their charges' value and electrostatic potential in the vicinity of the membrane surface can be approximated by the Guy-Chapman theory. This theory treats the interface as a charged structureless plain surrounded by uniform environments. However, if one considers intermolecular interactions, such assumptions need to be revised. The interface is in reality a thick region containing the residual charges of lipid headgroups. Their arrangement depends on the type of lipid present in the membrane. The variety of lipids and their biological functions suggests that charge distribution determines the extent and type of interaction with surface associated molecules. Numerous examples show that protein behavior at the lipid bilayer surface is determined by the type of lipid present, indicating protein specificity towards certain surface locations and local properties (determined by lipid composition) of a particular type. Such specificity is achieved by a combination of electrostatic, hydrophobic and enthropic effects. Comparing lipid biological activity, it can be stated that residual charge distribution is one of the factors of intermolecular recognition leading to the specific interaction of lipid molecules and selected proteins in various processes, particularly those involved with signal transduction pathways. Such specificity enables a variety of processes occurring simultaneously on the same membrane surface to function without cross-reaction interference.

Animals↗

Reconstruction of the mandible by prefabricated autogenous bone grafts. An experimental study in minipigs.

The aim of the present experimental study was to evaluate the use of prefabricated bone grafts for reconstruction of the mandible. In 20 adult Göttingen minipigs, prefabricated bone grafts 10 x 12 x 40 mm in size were cultivated in scaffolds of pyrolized bovine bone under a polylactic membrane on the outside of the mandible during a period of five months. The grafts were harvested and transferred to bridge 2- and 4-cm lower mandibular border defects and discontinuity defects. Five animals served as ungrafted controls and evaluation of the grafts was performed three and five months after grafting. At both intervals and in both graft-length groups, the grafted bone volume was almost completely preserved inside the scaffolds and exhibited a slight (3 months) to marked (5 months) increase in bone density by appositional bone growth. The inserted screws were histologically integrated into the transplanted bone and the grafts were linked to the adjacent mandibular bone without intervening soft tissue. The grafts, which were transferred to bridge discontinuity defects, were likewise well preserved with direct fusion between the grafted bone and local bone. It was concluded that bone grafts can be prefabricated from underlying mandibular bone and used for the repair of mandibular defects of various length and shape.

Animals↗

Dietary fatty acids alter the adhesion properties of lymphocytes to extracellular matrix proteins.

Dietary fats have been shown by many investigators to affect immune responses in vitro and in vivo. However, the exact mechanism(s) by which fats or their metabolic derivatives affect immune function is still unknown. In this report we have investigated whether short-term in vitro exposure to fatty acids alters the adhesion of lymphocytes to extracellular matrix proteins. We found remarkably heterogeneous effects with these agents on lymphocyte adhesion; increases and decreases in adhesion were observed depending upon the fatty acid, cell type and extracellular matrix protein used. Alterations in the adhesion potential of lymphocytes could serve as a mechanism for the reported effects of fatty acids on immune function since lymphocytes are dependent upon rapid and reversible adherence events for most of their effector activities.

Animals↗

The role of helper lipids in cationic liposome-mediated gene transfer.

In the procedure for cationic liposome-mediated transfection, the cationic lipid is usually mixed with a "helper lipid" to increase its transfection potency. The importance of helper lipids, including dioleoylphosphatidylcholine (DOPC) and phosphatidylethanolamine (dioleoyl PE), DO was examined. Freeze-fracture electron microscopy of DNA:cationic complexes containing the pSV-beta-GAL plasmid DNA, the cationic lipid dioleoyl trimethylammonium propane, and these helper lipids showed that the most efficient mixtures were aggregates of ensheathed DNA and fused liposomes. PE-containing complexes aggregated rapidly when added to culture media containing polyanions, whereas PC-containing complexes did not. However, more granules of PC-containing complexes were formed on cell surfaces after the complexes were added to Chinese hamster ovary (CHO) cells in transfection media. Pronase treatment inhibited transfection, whereas dilute poly-L-lysine enhanced transfection, indicating that the attachment of DNA:liposome complexes to cell surfaces was mediated by electrostatic interaction. Fluorescence spectroscopy studies confirmed that more PC-containing complexes than PE-containing complexes were associated with CHO cells, and that more PC-containing complexes were located in a low pH environment (likely to be within endosomes) with time. Cytochalasin-B had a stronger inhibitory effect on PC-containing liposome-mediated than on PE-containing liposome-mediated transfection. Confocal microscopic recording of the fluorescently label lipid and DNA uptake process indicated that many granules of DNA:cationic liposome complexes were internalized as a whole, whereas some DNA aggregates were left out on the cell surfaces after liposomes of the complexes fused with the plasma membranes. For CHO cells, endocytosis seems to be the main uptake pathway of DNA:cationic liposome complexes. More PC-containing granules than PE-containing granules were formed on cell surfaces by cytoskeleton-directed membrane motion, after their respective DNA:liposome complexes attached to cell surfaces by electrostatic means. Formation of granules on the cell surface facilitated and/or triggered endocytosis. Fusion between cationic liposomes and the cell membrane played a secondary role in determining transfection efficiency.

Animals↗

Interaction of free fatty acids with phospholipid bilayers.

The partition of free fatty acids (FFA) to egg-phosphatidylcholine (egg-PC) and egg-phosphatidylethanolamine (egg-PE) vesicles was studied. Upon the addition of FFA to the suspension of vesicles, the pH of the aqueous phase changed depending on the length and saturation of the FFA hydrocarbon chain, as well as on the vesicle composition. The medium pH decreased faster if FFA was added to egg-PE as compared to egg-PC vesicles. The fluorescent free fatty acid indicator (ADIFAB) was used to measure the amount of FFA remaining in the aqueous phase. Most of the FFA added to the suspension of egg-PE vesicles remained in the aqueous phase, whereas in the presence of egg-PC vesicles the FFA partitioned preferentially into the lipid phase. The amount of FFA incorporated into the lipid bilayers was estimated by measuring the changes of pH at the lipid bilayer surface, using fluorescein-PE. At high surface concentrations of FFA, decreasing pH at the bilayer surface caused the protonation of FFA, and raised the pK of FFA at the bilayer surface from 5 to about 7. The partition of FFA in egg-PE vesicles was an order of magnitude lower than that in egg-PC vesicles. The incorporation amount was determined more by the molecular packing than by the nature of lipid headgroups, because steroylcaprioyl-PE, which preferred the bilayer structure, behaved more like egg-PC than egg-PE. Understanding FFA partition characteristics would help to interpret the hydrolysis measurements of phospholipids, and to explain many biological activities of FFA.

Fatty Acids, Nonesterified↗

Merocyanine interaction with phosphatidylcholine bilayers.

Merocyanine (MC 540) is a fluorescent probe whose optical properties depend on the environmental polarity. In the presence of lipid bilayers, MC 540 binds to the membrane surface while simultaneously changing its fluorescence properties. Previous studies have shown that the fluorescence of merocyanine depends upon the lipid packing in the membrane. We measured the partitioning of MC 540 and its fluorescence properties in the presence of phosphatidylcholine membranes. We found that the fluorescence of MC 540 shows, as expected, a major change around the main phase transition of phosphatidylcholine membranes. However, instead of a step-like increase of fluorescence, the maximum at phase transition was observed. We were able to explain our data by combining two effects; dependence of MC 540 fluorescence on temperature and lipid fluidity. In addition, we established that the increase of the fluorescence intensity in the presence of lipid bilayers in the fluid state is due to the elevated partitioning of the probe into the lipid phase. The partition of MC 540 into the fluid membrane does not depends on the dye concentration in the aqueous phase. When lipid was in the gel phase the partitioning of the dye increased with its bulk concentration, whereas the fluorescence intensity remained unchanged. We conclude, therefore, that MC 540 forms nonfluorescent complexes when in the gel lipid membrane.

Chemical Phenomena↗

Dithionite penetration through phospholipid bilayers as a measure of defects in lipid molecular packing.

The permeability of dithionite through bilayers was utilized to probe the structural defects in the bilayers of these lipids through their respective gel-fluid and bilayer-hexagonal phase transitions. The water soluble dithionite ion penetrates intact bilayers very slowly. The rate of irreversible quenching of the fluorescence of NBD-PE labelled liposomes may thus be used as an indicator of the permeability of this ion through bilayers. The quenching rate has a fast and a slow component, the fast one corresponds to the quenching of fluorophores immediately accessible to the quencher, i.e. those on the outer surface of liposomes. The slower component represents the average rate of penetration of the quencher through the bilayer, to quench those fluorophores at the inner shells of the multilamellar vesicles. Both rates may be approximated by a single exponential function. The slow exponent is simply related to the permeability. The permeability of DMPC as a function of temperature shows a peak at the gel-fluid phase transition at 24 degrees C, but returns to about the pre-transition value at temperatures above the phase transition. The permeability of egg PE shows a hump at 45 degrees C before the hexagonal phase transition at 65 degrees C is reached and becomes infinite at the hexagonal phase transition as all fluorophores are immediately accessible to the quencher. We believe that the permeability measured by this method relates more to the molecular packing defects which maximizes at the gel-fluid phase transition temperatures just below the bilayer-hexagonal phase transition, rather than the general packing order which simply changes with structural phases.

Cell Membrane Permeability↗

[Three cases of thoracic spina fracture with displacement without spinal cord injury].

Three cases of extremely rare, thoracic spine displaced fractures without injury of the spinal cord are reported. An attempt to reconstruct injury mechanism and pathology of the fracture has been undertaken on the basis of radiographs analysis. Immobilisation treatment with no reduction yielded good results. The authors are convinced that this paper, if studied, will indicate the management of this type of injury to those come across it for the first time.

Adolescent↗

Relationship between membrane lipid mobility and spectrin distribution in lymphocytes.

We have previously established that T and B lymphocytes in situ are remarkably heterogeneous with respect to the cytoskeletal protein spectrin. Since in erythrocytes spectrin is known to play an important role in the regulation of membrane fluidity, lipid organization and lateral mobility of membrane proteins, we have sought to determine if the heterogeneous patterns of spectrin distribution that we have observed are related to possible differences in membrane lipid organization in these various subsets. To this end, we have utilized a fluorescent pyrene-labelled phospholipid as a probe of the lipid lateral mobility and have examined two related T cell systems maintained in vitro, DO.11.10 cells and a spontaneously arising variant, DO.11.10V. In these (and other cloned in vitro systems) we have previously observed that the cells homogeneously express one of the kinds of spectrin distribution patterns observed in situ. Thus the uniformity of staining of these systems permits us to address whether the various patterns of spectrin distribution may be predictive of differences in membrane lipid properties. Here we show that in cells in which there is little or nor spectrin at the plasma membrane (DO.11.10) that the lipids in the plasma membrane are considerably less mobile than in its related variant in which spectrin is diffusely distributed within the cell and at the plasma membrane. From this and previous results, we conclude that differences in the distribution of the cytoskeletal protein spectrin among lymphocytes may be a useful parameter in helping to predict the status of membrane lipid organization.

Cell Line↗

Iodide penetration into lipid bilayers as a probe of membrane lipid organization.

The quenching efficiency of iodide as a penetrating fluorescence quencher for a membrane-associated fluorophore was utilized to measure the molecular packing of lipid bilayers. The KI quenching efficiency of tryptophan-fluorescence from melittin incorporated in DMPC bilayer vesicles peaks at the phase transition temperature (24 degrees C) of DMPC, whereas acrylamide quenching efficiency does not depend on temperature. The ability of iodide to penetrate the hydrocarbon region of the bilayer was examined by measuring the fluorescence quenching of the pyrene-phosphatidylcholine incorporated into DMPC vesicles (pyrene was attached to the 10th carbon of the sn-2 chain). The quenching efficiency of pyrene by iodide again shows a maximum at the lipid phase transition. We conclude that iodide penetrates the membrane hydrocarbon region at phase transition through an increased number of bilayer defects. The magnitude of change in quenching efficiency of iodide during lipid phase transition provides a sensitive technique to probe the lipid organization in membranes.

Lipid Bilayers↗