Effect of sulfadiazine on biological model membranes

Summary In this paper the optimization of the kaolin calcination is studied, aiming at using the produced metakaolin as supplementary cementitious material. Representative samples of poor Greek kaolin (Milos island) and a high purity commercial kaolin were tested. Samples were heated at different temperatures during different times. The optimization of calcination conditions was studied by DTA-TG and XRD analysis of the raw and thermal treated kaolin samples, by pozzolanic activity analysis of metakaolins and finally by strength development analysis of cement-metakaolin mixtures. This approach showed that heating at 650°C for 3 h is efficient to convert poor kaolins with low alunite content to highly reactive metakaolins. However in the case of kaolin with a high alunite content, thermal treatment at 850°C for 3 h is required in order to remove undesirable SO₃. Evidence was found that poor kaolins can be efficiently used for the production of highly reactive metakaolins.     

Wetting properties of model biological membranes

Various aspects of native and model biological membrane wettability are discussed. Among others hydration of mono-, bi-, and multi-layers of lipids as well as wettability of macroscopic surfaces of solid supported lipid films was investigated via apparent contact angle measurements and calculation of the apparent surface free energy of the films. The effects of relative humidity on the layer hydration and contact angle changes are also discussed. Finally, the effect of liposomes and enzymes (due to the hydrolysis reactions) on the hydrophobic/hydrophilic character of the film surfaces is overviewed.     

Studies on non-thiazolidinedione antidiabetic agents. 3. Preparation and biological activity of the metabolites of TAK-559

Preparation and biological activity of the metabolites of the potent antihyperglycemic and antihyperlipidemic agent, (E)-4-(4-[(5-methyl-2-phenyl-1,3-oxazol-4-yl)methoxy]benzyloxyimino)-4-phenylbutyric acid (TAK-559) (1), were investigated. Metabolites M-I (2), M-II (3), M-III (4) and M-IV (5) were synthesized and their biological activities were evaluated by in vitro and in vivo experiments. Compounds 2-4 activate human peroxisome proliferator-activated receptor gamma one (hPPARgamma1) and hPPARalpha, but their activities are weaker than those of TAK-559 (1). Compound 5 only activates hPPARgamma1 weakly. TAK-559 (1) showed potent in vivo plasma glucose and triglyceride lowering activities in Wistar fatty rats after intraperitoneal administration, while its metabolites (2-5) showed comparatively weak activities.     

Fluorescence lifetimes study of alpha-tocopherol and biological prenylquinols in organic solvents and model membranes

We have found that for biological prenyllipids, such as plastoquinol-9, alpha-tocopherol quinol, and alpha-tocopherol, the shortest fluorescence lifetimes were found in aprotic solvents (hexane, ethyl acetate) whereas the longest lifetimes were those of ubiquinonol-10 in these solvents. For all the investigated prenyllipids, fluorescence lifetime in alcohols increased along with an increase in solvent viscosity. In a concentrated hexane solution, the lifetimes of prenylquinols considerably decreased. This contrasts with methanol solutions, which is probably due to the self-association of these compounds in aprotic solvents. We have also found a correlation of the Stokes shift of prenyllipids fluorescence with the orientation polarizability of the solvents. Based on data obtained in organic solvents, measurements of the fluorescence lifetimes of prenyllipids in liposomes allowed an estimation of the relative distance of their fluorescent rings from the liposome membrane surface, and was found to be the shortest for alpha-tocopherol quinol in egg yolk phosphatidylcholine liposomes, and increased in the following order: alpha-tocopherol in dipalmitoyl phosphatidylcholine liposomes < alpha-tocopherol < plastoquinol-9 < ubiquinol-10 in egg-yolk phosphatidylcholine liposomes.     

Studies on bio-antioxidants II. An ESR study on the antioxidant efficiency of ascorbyl palmitate in micelles

The kinetics for the reaction of ascorbyl palmitate and 4-hydroxy-TEMPO, 4-methoxy-TEMPO or 4-hexanoyloxy-TEMPO in CTAB and SDS micelles was studied by ESR spectroscopy. It was found that the behavior of ascorbyl palmitate is remarkably different from ascorbic acid in their antioxidant activity in micelles. The predominant factors governing the activity in micelles involve the lipophilicity of the antioxidant and the oxidant, the charge type of the substrates and the concentration of the surfactant. As high as 2 × 10⁴ fold rate variation was observed by changing the lipophilicity of the antioxidant and the microenvironment of the reaction medium.     

The interaction of helical polypeptides with biological model membranes

Proton-decoupled solid-state ¹⁵N NMR spectroscopy was used to investigate helical peptides reconstituted into oriented phospholipid bilayers. Hydrophobic channel peptides such as the N-terminal region of Vpu of human immunodeficiency virus (HIV-1) adopt transmembrane orientations, whereas amphipathic peptide antibiotics are oriented parallel to the bilayer surface. The alignment of helical peptides in lipid membranes was analysed in some detail using model peptides. In particular, peptides with pH-dependent topology and a series of peptides that allow one to study the contributions of specific interactions were designed. The energies of transfer of several amino acids from the in-plane to transmembrane localisation were determined. In addition, the alignment of peptides and phospholipids under conditions of hydrophobic mismatch have been investigated in considerable detail.     

Increased antioxidant reactivity of vitamin C at low pH in model membranes

The reactivity of the water-soluble antioxidant vitamin C (l-ascorbic acid) depends on pH. It is generally recognized that the ascorbate monoanion, which predominates at neutral physiological pH, acts as a stronger antioxidant than the protonated form, ascorbic acid. Fluorazophore-L, a long-lived fluorescent probe, was employed as a mimic for lipid peroxyl radicals. The experiments with micellar and bilayer membrane models demonstrate that vitamin C becomes, in fact, a more powerful antioxidant at low pH. This phenomenon may be general for the interception of reactive oxidizing species at the lipid/water interface.     

Spatial distribution of ion channel activity in biological membranes: the role of noise

A new approach is proposed to model a collective ion channel dynamics. We have assumed that ion channels create a two-component spatio-temporal interaction field. Every channel at its current spatial location in membrane contributes permanently to this field with its state (open or closed) and coupling strength to other channels. This field is described by a reaction-diffusion equation, the transition of ion channel from closed to open state (and vice versa) is described by a master equation, and migration of channels in membrane is described by a set of Langevin equations coupled by the interaction field. Within this model, we have investigated critical conditions for spatial distribution of ion channel activity.     

Synthesis of TAK-218 using (R)-2-methylglycidyl tosylate as a chiral building block

We performed an asymmetric synthesis of (S)-2,3-dihydro-2,4,6,7-tetramethyl-2-[(4-phenyl-1-piperidinyl)methyl]-5-benzofuranamine dihydrochloride (TAK-218, 1), a compound used for the treatment of traumatic and ischemic central nervous system injuries. Oxirane 6, which was synthesized from (R)-2-methylglycidyl tosylate, was treated with aqueous trifluoroacetic acid to afford benzofuranmethanol 7 with inversion of stereochemistry at the stereogenic center. Compound 7 was converted into 1 with high enantiomeric excess in four steps.     

Distribution of fullerene nanomaterials between water and model biological membranes

Biological membranes are one of the important interfaces between cells and pollutants. Many polar and hydrophobic chemicals can accumulate within these membranes. For this reason, artificial biological membranes are appealing surrogates to complex organisms for assessing the bioaccumulation potential of engineered nanomaterials (ENMs). To our knowledge, this work presents the first quantitative study on the distribution of fullerene ENMs between lipid bilayers, used as model biological membranes, and water. We evaluated the lipid bilayer-water association coefficients (K(lipw)) of aqueous fullerene aggregates (nC(60)) and fullerol (C(60)(ONa)(x)(OH)(y), x + y = 24). Kinetic studies indicated that fullerol reached apparent equilibrium more rapidly than nC(60) (2 h versus >9 h). Nonlinear isotherms can describe the distribution behavior of nC(60) and fullerol. The lipid bilayer-water distributions of both nC(60) and fullerol were pH-dependent with the accumulation in lipid bilayers increasing systematically as the pH decreased from 8.6 (natural water pH) to 3 (the low end of physiologically relevant pH). This pH dependency varies with the zeta potentials of the ENMs and leads to patterns similar to those previously observed for the lipid bilayer-water distribution behavior of ionizable organic pollutants. The K(lipw) value for nC(60) was larger than that of fullerol at a given pH, indicating a greater propensity for nC(60) to interact with lipid bilayers. For example, at pH 7.4 and an aqueous concentration of 10 mg/L, K(lipw) was 3.5 times greater for nC(60) (log K(lipw) = 2.99) relative to fullerol (log K(lipw) = 2.45). Comparisons with existing aquatic organism bioaccumulation studies suggested that the lipid bilayer-water distribution is a potential method for assessing the bioaccumulation potentials of ENMs.     

Using ESR spectroscopy to study radical intermediates in proton-exchange membranes exposed to oxygen radicals

Direct ESR and spin-trapping experiments were used to study the behavior of Nafion, a perfluorinated ionomer membrane used in fuel cells, when exposed in the laboratory to oxygen radicals produced by Fenton and photo-Fenton reactions. DMPO (5,5-dimethyl-1-pyroline) was used as the spin trap. The results suggest that the two ESR methods provide complementary information on Nafion fragmentation. The presence of membrane-derived fragments was suggested indirectly by the presence of a broad signal (line width ≈ 84 G) after prolonged exposure of the membrane to the Fenton reagent based on Ti(III), and by the DMPO adduct of a carbon-centered radical in the spin-trapping experiments. The most convincing proof for the presence of perfluorinated radicals was obtained in Nafion membranes partially neutralized by Cu(II), Fe(II) and Fe(III) upon exposure to UV-irradiation in the presence or absence of H₂O₂ (photo-Fenton treatment). Identification of the chain-end radical RCF₂CF ₂ ^• with magnetic parameters different to those determined for the chain-end detected in γ-irradiated Teflon, was taken as evidence for the attack of reactive oxygen radicals on the side-chain of the membrane. Additional support for this suggestion was the detection of the “quartet” ESR signal assigned to the CF₃C^•O radical, and of the “quintet” ESR signal assigned to the radical centered at the intersection of the main and side chains. The limitations and advantages of each approach are discussed.     

Elastic and flexoelectic aspects of out-of-plane fluctuations in biological and model membranes

Out-of-plane fluctuations of biological membranes representing an important manifestation of cell surface dynamics were discovered about one hundred years ago, but their purely physical (Brownian) character has been recognized only recently. This has been a demonstration of the successful application of the liquid crystal approach to the structure and functions of biomembranes. The present article concentrates on the liquid crystal (elastic and flexoelectric) aspects of these fluctuations of membrane curvature. The theoretical basis of curvature elasticity and curvature-induced (flexoelectric) polarization of a membrane is provided by liquid crystal physics. Special attention is paid to experimental and theoretical studies of curvature elasticity and flexoelectricity of a pure lipid bilayer in a liquid crystal state.

Optical and electrical (noise) investigations of out-of-plane fluctuations in model and living membrane systems are reviewed. The influence of out-of-plane fluctuations on surface interactions and cell-cell contact of flexoelectric membranes is considered. The article concludes by underlining the unique character of information about the mechano-electrical and other liquid crystal properties of membranes that may be gained from future extensive studies of membrane fluctuations.     

ESR study on the photosensitized decomposition of polyethylene

Several aromatic compounds were added to low-density polyethylene to determine their effects on the photodegradation of polyethylene. It was shown that some aromatic compounds indeed sensitize the hydrogen abstraction of the allylic hydrogen of unsaturated groups contained in polyethylene as an improper bond. The mechanism of photosensitization was investigated by an ESR method. It was found that the higher photoexcited triplet state of an aromatic molecule produced by the biphotonic ultraviolet light absorption transfers its excess energy to the unsaturated bond to excite it, and the excited unsaturated group releases its allylic hydrogen atom, giving an allylic radical.     

ESR and electrochemical study of 5-nitroindazole derivatives with antiprotozoal activity

The electrochemistry of 3-alkoxy- and 3-hydroxy-1-[omega-(dialkylamino)alkyl]-5-nitroindazole derivatives were characterized using cyclic voltammetry in DMSO. The nitro reduction process was studied and this was affected by the acid moieties present in these compounds. A nitro anion self-protonation process was observed. This phenomenon was studied by cyclic voltammetry in presence of increasing amount of NaOH. The reactivity of the nitro anion radical of these derivatives with glutathione was also studied by cyclic voltammetry. The oxidizing effect of glutathione is supported by the parallel decrease of the anodic peak current and increase of the cathodic peak in the cyclic voltammograms, corresponding to the wave of the nitro anion radical from uncharged species with the addition of glutathione. Nitro anion radicals obtained by electrolytic reduction of these derivatives were measured and analyzed in DMSO using electron spin resonance spectroscopy.     

Towards a minimal stochastic model for a large class of diffusion-reactions on biological membranes

Diffusion of biological molecules on 2D biological membranes can play an important role in the behavior of stochastic biochemical reaction systems. Yet, we still lack a fundamental understanding of circumstances where explicit accounting of the diffusion and spatial coordinates of molecules is necessary. In this work, we illustrate how time-dependent, non-exponential reaction probabilities naturally arise when explicitly accounting for the diffusion of molecules. We use the analytical expression of these probabilities to derive a novel algorithm which, while ignoring the exact position of the molecules, can still accurately capture diffusion effects. We investigate the regions of validity of the algorithm and show that for most parameter regimes, it constitutes an accurate framework for studying these systems. We also document scenarios where large spatial fluctuation effects mandate explicit consideration of all the molecules and their positions. Taken together, our results derive a fundamental understanding of the role of diffusion and spatial fluctuations in these systems. Simultaneously, they provide a general computational methodology for analyzing a broad class of biological networks whose behavior is influenced by diffusion on membranes.     

Multiquantum processes in the enzyme molecules immobilized on biological membranes

A physical model of an oxidation–reduction reaction for immobilized enzyme is considered. The influence of the electric-field intensity of a biological membrane on the enzyme reaction rate is analyzed. It is shown that the low-frequency dipole-active vibration of the oppostite charge groups of the substrate and enzyme relative to each other leads to multiquantum excitation of the system over the conformational degree of freedom. This excitation provides an abrupt increasing of the tunnel decay rate of the substrate–enzyme complex on a free product and a free enzyme. This way of the reaction is more probable in comparison with the usual overbarrier process. Some consequences for the immobilized cytochrome P-450 are discussed. © 1998 John Wiley & Sons, Inc. Int J Quant Chem 66 : 255–260, 1998     

Organization of phospholipids in biological membranes

Preferred conformations of phospholipids have been predicted through quantum-chemical techniques and classical potential functions. An essential condition for a conformation to exist in a biomembrane is that it should be possible for it to organize in the form of a bilayer. Taking into consideration the conformational flexibility of the polar head group, organization at the lipid–water interface has been considered. The biological implications of such an organization in terms of formation of “hydrophobic channels” is discussed. Quantum-mechanical investigations on the transport phenomenon have shown that the “selectivity” of biological membranes is connected with the “organization.” Calculations of the quantum-mechanical transmission coefficients for different model potential profiles indicate that minor differences in the height of potential barriers in certain regions can lead to significant changes in transmission coefficients. The “directional selectivity” of substrates (differences in transmission coefficients for flow in and out of the cell) can be explained on the basis of differences in membrane organization. These results have some important consequences in the evolutionary process in biological membranes.     

A theoretical study on the antioxidant activity of Uralenol and Neouralenol scavenging two radicals

Uralenol and neouralenol are two typical licorice root extracts that presents multiple reactive hydroxyl groups, which are considered as good free radical scavengers. A theoretical study on the primary antioxidant activity of uralenol and neouralenol toward hydroxyl and hydroperoxyl radicals has been carried out using the density functional theory (DFT). A total of 10 reaction pathways of uralenol and neouralenol scavenging two radicals in gas phase and in water phase have been tracked. Neouralenol was found to be a better hydroxyl and hydroperoxyl scavenger than uralenol. In vivo, the more reactive sites in uralenol are U5 and U’1, respectively, for scavenging ·OH and ·OOH; and the more reactive sites in neouralenol are N4 and N’5 for scavenging ·OH and ·OOH, respectively.