Helmut Ringsdorf

Domain Structures in Langmuir-Blodgett Films Investigated by Atomic Force Microscopy

Investigations of phase-separated Langmuir-Blodgett films by atomic force microscopy reveal that on a scale of 30 to 200 micrometers, these images resemble those observed by fluorescence microscopy. Fine structures (less than 1 micrometer) within the stearic acid domains were observed, which cannot be seen by conventional optical microscopic techniques. By applying the force modulation technique, it was found that the elastic properties of the domains in the liquid condensed phase and grains observed within the liquid expanded phase were comparable. Small soft residues in the domains could also be detected. The influence of trace amounts of a fluorescence dye on the micromorphology of monolayers could be detected on transferred films.

Hydrolytic action of phospholipase A2 in monolayers in the phase transition region: direct observation of enzyme domain formation using fluorescence microscopy

Phospholipase A2, a ubiquitous lipolytic enzyme highly active in the hydrolysis of organized phospholipid substrates, has been characterized optically in its action against a variety of phospholipid monolayers using fluorescence microscopy. By labeling the enzyme with a fluorescent marker and introducing it into the subphase of a Langmuir film balance, the hydrolysis of lipid monolayers in their liquid-solid phase transition region could be directly observed with the assistance of an epifluorescence microscope. Visual observation of hydrolysis of different phospholipid monolayers in the phase transition region in real-time could differentiate various mechanisms of hydrolytic action against lipid solid phase domains. DPPC solid phase domains were specifically targeted by phospholipase A2 and were observed to be hydrolyzed in a manner consistent with localized packing density differences. DPPE lipid domain hydrolysis showed no such preferential phospholipase A2 response but did demonstrate a preference for solid/lipid interfaces. DMPC solid lipid domains were also hydrolyzed to create large circular areas in the monolayer cleared of solid phase lipid domains. In all cases, after critical extents of monolayer hydrolysis in the phase transition region, highly stabile, organized domains of enzyme of regular sizes and morphologies were consistently seen to form in the monolayers. Enzyme domain formation was entirely dependent upon hydrolytic activity in the monolayer phase transition region and was not witnessed otherwise.

An enzyme caught in action: Direct imaging of hydrolytic function and domain formation of phospholipase A2 in phosphatidylcholine monolayers

Phospholipase A2, a ubiquitous lipolytic enzyme that actively catalyses hydrolysis of phospholipids, has been studied as a model for enzyme‐substrate reactions, as a membrane structural probe, and as a model for lipid‐protein interactions. Its mechanism of action remains largely controversial. We report here for the first time direct microscopic observation of the lipolytic action of fluorescently marked phospholipase A2 (Naja naja naja) against phosphatidylcholine monolayers in the lipid phase transition region. Under these conditions, phospholipase A2 is shown to target and hydrolyse solid‐phase lipid domains of L‐α‐dipalmitoylphosphatidylcholine. In addition, after a critical extent of monolayer hydrolysis, the enzyme itself aggregates into regular, visible proteinaceous domains within the lipid monolayer. Solid‐phase lipid hydrolysis indicates a preferential hydrolytic environment for phospholipase A2 while enzyme domain formation points to a possible allosteric inhibition mechanism by hydrolysis products.

Second-harmonic generation in Langmuir–Blodgett monolayers of stilbazium salt and phenylhydrazone dyes

The second-order nonlinear optical susceptibilities χ(2) of several phenylhydrazone and stilbazium salt dyes in Langmuir–Blodgett monolayers have been determined from second-harmonic-generation measurements. Three of the substances demonstrated χ(2) values greater than 10−6 electrostatic units, although two of the three did not absorb light significantly at the second-harmonic wavelength.

Polymeric monolayers and liposomes as models for biomembranes

Polymer chemists as poachers in foreign grounds? Why not? Macromolecular chemistry has become a mature science with all advantages and handicaps of a well-established scientific discipline: many heights are conquerred and the harvest is abundant, but adventures and the future might be elsewhere. Besides, in these times of bottomed out industrial profits in common plastics, future polymer chemistry cannot be limited to repetitive improvement of already successful mass polymers but should rather expand into neighboring fields of material science as well as life science where “polymer thinking” might help to overcome difficulties. — First hesitant steps on the bridge towards membrane biology have been made.

Reduced protein adsorption on plastics via direct plasma deposition of triethylene glycol monoallyl ether

The direct plasma-induced deposition of tri(ethylene glycol) monoallyl ether is reported. RF plasma polymerization of this monomer was carried out under both continuous wave (CW) and pulsed plasma operation. The major focus of this work was optimization of the degree of retention of the C-O-C bonds of the starting monomer during the deposition process. This successfully was accomplished using low RF power during the CW runs and low RF duty cycles during the pulsed plasma experiments. Spectroscopic analysis of the plasma films revealed a strong dependence of film composition on the RF power and duty cycles employed. In particular, an unusually high level of film chemistry compositional control was demonstrated for the pulsed plasma studies, with film composition varying in a steady, progressive fashion with sequential changes in the ratios of plasma on to plasma off times. This film chemistry controllability is demonstrated despite the relatively low volatility of the starting monomer. The utility of this plasma deposition approach in introducing polyethylene oxide (PEO) structures on solid substrates was evaluated via protein adsorption studies. Radiolabeled bovine albumin adsorption was studied on plasma-modified poly(ethylene teraphthalate) (PET) substrates. Dramatic reductions in both initial adsorption and retention of this protein were observed on PET samples having maximal PEO content relative to its adsorption on untreated PET surfaces. Good stability and adhesion of the plasma films to the underlying PET substrates were observed, as evidenced from prolonged immersion of plasma-treated surfaces in aqueous solution. Overall, the results obtained from the present work provide additional support for the utility of one-step plasma process to reduce biological fouling of surfaces via deposition of PEO surface units.

Self-organization of substituted azacrowns based on their discoid and amphiphilic nature

Abstract Cyclame and hexacyclene derivatives, bearing four and six long-chain substituents respectively, were synthesized. They are discussed as monolayer-forming amphiphiles as well as liquid-crystalline-phase-forming thermotropic mesogens. The compounds investigated form ordered monolayers at the gas/water interface. In the monolayer the hydrophilic cyclic head group lies flat on the water surface, whereas the hydrophobic substituents are oriented perpendicularly with respect to the interface. Most derivatives fitted with aromatic substituents exhibit a solid condensed state exclusively. In contrast with this, solid condensed as well as expanded phases can be found when spreading the aliphatic-substituted compounds. In the latter case, the onset of the phase transition takes a bump-like shape, owing to kinetic reasons. A liquid-crystalline columnar order is only achieved with hexacyclenes bearing aromatic substituents, etherified with one alkyl chain. Besides this, the remaining derivatives melt from th...

Polymer therapeutics—polymers as drugs, drug and protein conjugates and gene delivery systems: Past, present and future opportunities

Welsh School of Pharmacy, Centre for Polymer Therapeutics, Cardiff University, Redwood Building, King Edward VII Avenue, Cardiff CF10 3XF, UK, University of Mainz, Institute of Organic Chemistry, Duesbergweg 10-14 D-55099, Mainz, Germany, and Vascular Biology Program, Department of Surgery, Children’s Hospital Boston and Harvard Medical School, 1 Blackfan Circle, Karp Family Research Laboratories, Floor 12, Boston, MA 02115, USA

Induction of a nematic columnar phase in a discotic hexagonal ordered phase forming system

Abstract The induction of a nematic columnar phase in a discotic hexagonal ordered phase forming system is achieved by mixing hexakispentyloxytriphenylene 1 with a long chain derivative of trinitrofluorenone 3. The difference in chain length has a strong influence on the packing behaviour due to steric effects. The long hydrocarbon chains of the acceptor introduce a strong asymmetry into the electron donor acceptor complex. It could be shown by differential scanning calorimetry, optical microscopy and X-ray measurements that a nematic columnar phase is formed. In this mesophase the triphenylenes form columns but no hexagonal or orthorhombic lattice is built up. Each column behaves like a rod-like nematic mesogen. To prove that the long hexadecane alkyl chains of the acceptor are responsible for this induction, the acceptor 3 was mixed with the non-liquid-crystalline triphenylene derivative 2 containing six hexadecyloxy side groups. The long alkyl chains of the acceptor dissolve perfectly in the side chain...

Interactions of liposomes and hydrophobically-modified poly-(N-isopropylacrylamides): an attempt to model the cytoskeleton

The interactions of small unilamellar vesicles (SUV) and water-soluble copolymers were studied by fluorescence spectroscopy, differential scanning calorimetry (DSC) and quasi-elastic light scattering (QELS). The anchoring onto liposomal bilayer membranes of copolymers of N-isopropylacrylamide, N-(2-(1-naphthyl)ethyl)-N-n-octadecylacrylamide and or N-[4-(1-pyrenyl)butyl]-N-n-octadecylacrylamide (0.5 mol% of the octadecylacrylamide comonomer) was monitored by non-radiative energy transfer between excited naphthalene and pyrene. The anchoring process occurred on zwitterionic lecithin liposomes and on negatively charged phosphatidic acid liposomes, whether the bilayer was in the crystalline or the liquid-crystalline phase. Insertion of the copolymer octadecyl groups within crystalline bilayers was attributed to the presence of packing defects. Aqueous solutions of poly-(N-isopropylacrylamide) and of its hydrophobically-modified copolymers exhibit a lower critical solution temperature (LCST). The coil to globule collapse of the polymer chains which is known to occur as the aqueous solution is heated through the LCST, also took place when the copolymers were anchored onto vesicular bilayers. The copolymers remained anchored during this collapse and the liposomes were not destroyed. The process was thermo-reversible. Detailed aspects of the reversibility of the phenomenon depended on the relative values of the phase transition temperatures of the liposomes and of the polymer LCST.