Nanostructurated complexes of poly(beta,L-malate) and cationic surfactants: synthesis, characterization and structural aspects

Ionic complexes of microbially produced poly(beta,L-malic acid) and alkyltrimethylammonium surfactants with linear alkyl chains containing even numbers of carbon atoms from 14 up to 22, were investigated. Complexes with a stoichiometric or nearly stoichiometric composition were prepared by precipitation from equimolar mixtures of aqueous solutions of the two components. All complexes were found to adopt supramolecular stratified structures made of alternating layers of poly(beta,L-malate) and surfactant with a periodicity on the length scale of 3-5 nm, which increased proportionally to the length of the polymethylene chain. In these complexes, alkyl side chains with more than 16 carbon atoms were partially crystallized showing reversible melting at temperatures between 40 and 70 degrees C. After melting, a smectic LC phase that isotropicized at approximately 100 degrees C was observed for all of the complexes. Conformational and dimensional changes taking place in the complexes by effect of heating were analyzed by (13)C CP-MAS NMR and powder X-ray diffraction.     

Comblike poly(alpha-alkyl gamma-glutamate)s: computer simulation studies of an intermediate thermal phase

Monte Carlo (MC) simulations have been used to study the structure of an intermediate thermal phase of poly(alpha-octadecyl gamma,D-glutamate). This is a comblike poly(gamma-peptide) able to adopt a biphasic structure that has been described as a layered arrangement of backbone helical rods immersed in a paraffinic pool of polymethylene side chains. Simulations were performed at two different temperatures (348 and 363 K), both of them above the melting point of the paraffinic phase, using the configurational bias MC algorithm. Results indicate that layers are constituted by a side-by-side packing of 17/5 helices. The organization of the interlayer paraffinic region is described in atomistic terms by examining the torsional angles and the end-to-end distances for the octadecyl side chains. Comparison with previously reported comblike poly(beta-peptide)s revealed significant differences in the organization of the alkyl side chains.     

On the effect of surface active agents and their structure on the temperature-induced changes of normal and waxy wheat starch in aqueous suspension. Part I. Pasting and calorimetric studies

Pasting and calorimetric studies of normal and waxy wheat starch were performed in the presence of a series of ionic (sulphates, trimethyl ammonium bromides) and non-ionic (monoglycerides, maltosides) short (12 carbon atoms) and long (16 carbon atoms) n-alkyl chain surfactants. With the exception of the alkyl ammonium bromides, all of the short chain surfactants lower the pasting temperature (PT) in normal wheat starch, while the long chain surfactants have the opposite effect. Contrary, regardless of their chain length, all ionic surfactants lower the PT in waxy wheat starch while the non-ionic surfactants induce small, sometimes almost negligible changes in the PT. Calorimetric studies revealed the absence of a direct connection between the effect of surfactants on the onset of the starch gelatinization transition and the PT. However, in the presence of all surfactants, except the alkyl ammonium bromides, the PT of normal wheat starch was found to lie within or very close the temperature range within which the dissociation of the amylose–surfactant complexes takes place. Waxy wheat starch, in contrast, pasted at temperatures that fell within the temperature range of the starch gelatinization transition. This is taken as evidence of the existence of a correlation between the PT and the dissociation of the amylose–surfactant complexes.     

Crystal structures and thermal analyses of alkyl 2-deoxy-α-d-arabino-hexopyranosides

The crystal structures of alkyl 2-deoxy-α-d-arabino-hexopyranosides, with the alkyl chain lengths from C₈ to C₁₈, are established by the single crystal X-ray structural determination. The even-alkyl chain length derivatives crystallized orthorhombic, with space group P2₁2₁2₁, whereas the odd-alkyl chain length derivatives crystallized monoclinic, with space group P2₁. The sugar moieties retained a ⁴C₁ chair conformation and the conformation of the alkyl chains was all-trans. The molecules formed a bilayer structure, in which alkyl chains were interdigitated. The hydrogen bonds, originating from the sugar moieties, were observed in adjacent layers and also within the same layer, resulting in the formation of infinite chains. The alkyl chains arranged parallel to each other and formed planar structures. The thermal properties of the alkyl 2-deoxy glucosides were analyzed further. It was observed that none of the derivatives exhibited mesomorphism. This study establishes that the absence of the hydroxyl group at C-2 of the sugar moiety results in a non-mesogenic nature of the alkyl 2-deoxy-α-d-glycosides, as opposed to the profound mesogenic nature of the normal alkyl glycosides.     

Paramagnetic liposomes containing amphiphilic bisamide derivatives of Gd-DTPA with aromatic side chain groups as possible contrast agents for magnetic resonance imaging

Three amphiphilic DTPA bisamide derivatives containing long-chain phenylalanine esters (with 14, 16 and 18 carbon atoms in the alkyl chain) were synthesized and their corresponding gadolinium(III) complexes were prepared. The attempts to form paramagnetic micelles carrying the gadolinium(III) complexes yielded unstable or polydisperse micelles implying that the presence of the bulky aromatic side groups in the amphiphilic Gd-DTPA bisamide complexes results in an inefficient packing of the paramagnetic complex into micelles. All complexes were efficiently incorporated into liposomes consisting of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), yielding stable and monodisperse paramagnetic liposomes. All liposomes had a comparable size, typically between 120 and 160 nm. As a result of the reduced mobility of the gadolinium(III) complexes, solutions of these supramolecular structures show a higher relaxivity than solutions of Gd-DTPA. However, the relaxivity gain is lower compared to compounds consisting of purely aliphatic chains of the same length, most likely due to the less efficient packing or increased local mobility of the gadolinium(III) complex. In the case of the Gd-DTPA bisamide complex with 18 carbon atoms, the immobilization inside the liposomal structure is less effective, probably because the aliphatic chains of the complex are longer than the alkyl chains of the DPPC host, resulting in a relatively high local mobility. The paramagnetic liposomes containing the Gd-DTPA bisamide complexes with 14 carbon atoms showed the highest relaxivity because the optimal length match between the hydrophobic chains of the DPPC and the ligand allowed very efficient packing of the paramagnetic complex into the liposome.     

Identification of 1-alkyl-2-acyl-3-(2',3'-diacylglycerol)glycerols, a new type of lipid class, in harderian gland tumors of mice

A new class of alkyl glycerolipids, 1-alkyl-2-acyl-3-(2',3'-diacylglycerol)glycerols, was identified in lipid extracts prepared from harderian gland tumors of mice. After saponification, this lipid class yielded 1-alkyl-3-(1'-glycerol)glycerols. Identification was based on mass spectrometry, proton nuclear magnetic resonance spectroscopy, infrared spectroscopy, and chromatography of various derivatives and appropriate standards that were synthesized. The alkyl moieties of this unique lipid class consisted of saturated aliphatic chains with chain lengths of 14 to 20 carbon atoms. The acyl moieties were mostly saturated and monounsaturated aliphatic chains ranging from 14 to 24 carbon atoms. The alkyl and acyl moieties of 1-alkyl-2-acyl-3-(2',3'-diacylglycerol)glycerols were similar to those of alkyldiacylglycerols present in the same tissue, except for the presence of monounsaturated alkyl moieties in the latter. 1-Alkyl-2-acyl-3-(2', 3'-diacylglycerol)glycerols were only found in trace amounts in the normal harderian glands of mice. The total quantity of the alkyl and acyl moieties with a chain length greater than 20 carbon atoms in the alkyldiacylglycerols from tumors were considerably lower than those found in normal harderian glands of mice. This is the first report of the presence of bisglyceryl ether lipids in mammalian tissue; its unique chemical structure is consistent with the type of ether-linked lipid products that could be synthesized in the reaction catalyzed by alkyldihydroxyacetone-P synthase.     

Molecular dynamics study of complexes of poly(glutamate) and dodecyltrimethylammonium

The supramolecular organization of self-assembled stoichiometric complexes formed by dodecyltrimethylammonium cations and ionized poly(gamma-glutamic) acid has been investigated in chloroform solution. Atomistic molecular dynamics simulations have been performed considering three different starting conformations for the polyelectrolyte chain, two sets of electrostatic parameters, and two methods for evaluating the electrostatic interactions. Results indicate that the polypeptide chain tends to adopt an alpha-like helix conformation similar to that obtained in aqueous solution for the un-ionized poly(gamma-glutamic) acid. On the other hand, both surfactant...amide and surfactant...carboxylate interactions were identified in the complex, this multiple pattern being previously observed in other surfactant...polypeptide complexes. Although the influence of the force-field in the results has been found to be negligible, the method used to evaluate the electrostatic interactions affects significantly the dynamics of the system. The more important differences between the results obtained using the spherical cutoff and Particle Mesh Ewald methods are discussed.     

The role of hydrogen bonding in the cuticular wax of Hordeum vulgare L.

The role of hydrogen bonding in the cuticular wax of Hordeum vulgare L. has been investigated by comparing differential scanning calorimetry and X-ray powder diffraction results of the wax with those of n-alkane mixtures with chain-length distributions resembling that of the wax. It is concluded that hydrogen bonding prevents separation of the short and long chain-length distributions and results in the formation of an amorphous component which is large compared with that of a typical paraffinic wax. It seems that the longer ester chains (39 ≥n≥ 50) bridge the amorphous zone containing chain-ends between two adjacent layers of shorter chains (20 ≥n≥ 33), where n is the number of carbon atoms per chain. In contrast to a paraffinic wax, which has a monolayered structure, this plant wax has a bilayered structure.     

Interaction of human serum albumin with fatty acids. Role of anionic group studied by affinity partition.

The interaction of human serum albumin with fatty acids has been determined using the method of affinity partitioning in aqueous biphasic systems containing dextran, poly(ethylene glycol) and esters of dicarboxylic acids with poly(ethylene glycol). The difference in the partition of albumin in phase systems with and without the poly(ethylene glycol)-bound fatty acid group provides a measure of the interaction of fatty acids with the protein. The relative contribution of the polar and non-polar interaction to the binding of fatty acids to albumin has been estimated by comparing the present data with that obtained earlier using poly(ethylene glycol)-bound straight chain aliphatic hydrocarbons. In both cases, the aliphatic chain should contain a minimum of 8 carbon atoms to affect the partition of albumin and that the maximum effect is obtained with chains containing 16 carbon atoms. The effect of the polymer-bound fatty acid group is higher than the corresponding hydrocarbon only when the number of carbon atoms in it exceeds 12. The relative effect of polymer-bound 16-carbon chains, with and without a carboxyl group in the terminal position is independent of pH in the range 5--9.     

A molecular dynamics study of the stoichiometric complex formed by poly (alpha, L-glutamate) and octyltrimethylammonium ions in chloroform solution.

We present a molecular dynamics simulation at 300 K in explicit solvent environment of chloroform of the stoichiometric complex formed by poly(alpha,L-glutamate) and octyltrimethylammonium ions. We observed that the alpha-helix conformation of the polypeptide chain remains stable during a 2-ns run. The surfactant ions predominantly adopted an extended conformation that is stabilized by favorable interactions with the organic solvent. Analysis of the organization of the surfactant with respect to the polypeptide chain indicated that each octyltrimethylammonium cation was preferentially bound to more than one carboxylate group. It was found that the most populated arrangement was that with the surfactant cations interacting with two carboxylate groups simultaneously.     

The relationship between the structure of plastoquinone derivatives and their biological activity in Photosystem II of spinach chloroplasts

The relationship between the structure of reconstituted plastoquinone derivatives and their ability to recover the Hill reaction was investigated by extraction and reconstitution of lyophilized chloroplasts from spinach, followed by monitoring DCIP photoreduction at 600 nm. The results show that: It is not essential that the plastoquinone side chain be an isoprenoid or a phytol; the activity increases with increasing length of the side chain up to 13–15 carbon atoms; for chains longer than 15 carbon atoms, the activity is practically constant. Lipophilic groups (such as -Br) in the side chain increased the activity, hydrophilic groups (such as -OH) decreased the activity. Conjugated double bonds in the side chain decreased the activity greatly, but non-conjugated double bonds had almost no effect on the activity, indicating a requirement of flexibility of the side chain. The activity is decreased in the order of PQ, UbiQ and MQ, showing a large effect of the ring structure.Abbreviations DCIP 2,6-dichlorophenolindophenol - DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - Q_A primary electron acceptor in PS II reaction centers - Q_B secondary electron acceptor in PS II reaction centers - PQ _n plastoquinones with an isoprenoid side chain (n, number of the isoprenoid units in the side chain) - PQ-n synthetic plastoquinones with alkyl side chain (n, number of the carbon atoms in the alkyl side chain) - PQ-n synthetic plastoquinones with a double bond in the alkyl side chain - UQ _n ubiquinones with an isoprenoid side chain (n, number of the isoprenoid units in the side chain) - UQ-n synthetic ubiquinones with alkyl side chain (n, number of the carbon atoms in the akyl side chain) - MQ-n 2-alkyl-1,4-naphthoquinone (n, number of the carbon atoms in the alkyl side chain)     

The influence of the polar head and the hydrophobic chain on the skin penetration enhancement effect of poly(ethylene glycol) derivatives

The effect of a homologue series of nonionic surfactants, namely poly(ethylene glycol) (PEG) fatty acid esters, differing in oxyethylene (PEG 8, PEG 12, and PEG 40) and fatty acid (stearate, mono and di-laurate, and mono and di-oleate) chain lengths, on in vitro skin permeability of ketoprofen (KTP) vehicled in plasters was investigated. The drug diffusion through hairless mouse skin as well as the effect of the surfactant type and strength was studied by Franz diffusion cells and ATR-FTIR spectroscopy. The use of PEG stearate series revealed that the surfactant with the largest polar head, namely PEG 40, was ineffective in enhancing the skin permeation of KTP, independently of the plaster concentrations. The effect of the hydrophobic chain was investigated only by using the shortest oxyethylene chains. The experimental results revealed that the oxyethylene chain length of surfactants appeared to be more influent than the alkyl chain. The prediction of the absorption enhancing capability of these PEG derivatives appeared related to the vehicle other than the proper combination of the number of ethylene oxide groups and alkyl groups.     

Antibacterial and antifungal efficacy of surface functionalized polymeric beads in repeated applications

A simple method was developed to prepare polymeric microbeads with antibacterial and antifungal properties. The microbeads of approximately spherical shape and narrow size distribution were prepared from a mixture of poly (4-vinyl pyridine) (P4VP) and poly (vinylidene fluoride) (PVDF) by a phase inversion technique and subsequently derivatized with alkyl bromides having 4-10 carbon atoms. The quaternization of the pyridine groups into pyridinium groups confers the surface with highly effective and long-lasting antibacterial and antifungal properties, as shown by the effect on Escherichia coli and Aspergillus niger. Upon contact with the N-alkylated beads, the bacteria and fungal spores are lysed and intracellular constituents leach out into the medium. The efficacy of the alkyl chains in disrupting the cell membrane was investigated. The stability of the functional group and microbiocidal effectiveness of the microbeads in repeated applications was also assessed.     

Straight and branched (ω-1)-hydroxylated very long chain fatty acids are components of Bradyrhizobium lipid A

Lipopolysaccharides of seven Bradyrhizobium strains and three whole-cell fatty acid preparations from bacteria isolated from nodules of Sarothamnus scoparius (common broom) were studied for the presence of very long chain (ω-1)-hydroxy fatty acids. Several such fatty acids were identified. Among them, straight-chain as well as mono- and dimethyl branched acids with chains in the range from 26 to 34 carbon atoms were found. Pyrrolidides and 4,4-dimethyloxazoline derivatives were used to determine the branching position. Carbons at the (ω-10) and/or (ω-11) positions in alkyl chains were points of attachment of methyl groups. These data complete the structure of bradyrhizobial lipid A with important details. The obtained results can be applied in the chemotaxonomy of Bradyrhizobium.     

Minor and trace sterols in marine invertebrates : VI. Occurrence and possible origins of sterols possessing unusually short hydrocarbon side chains

Sterols with biosynthetically unusually short side chains (fewer than eight carbon atoms expected for primary squalene cyclization products) have been identified in the extracts of numerous marine invertebrates. The structures of the short side chain and conventional side chain sterols have been determined for various species of Porifera and Coelenterata. Sterol structures were determined by comparison of their mass spectra and gas chromatographic retention times with those of authentic or synthetic samples. Evidence is presented supporting the natural occurrence of these compounds in the tissues of the marine invertebrates as opposed to formation by degradative processes during sample handling or laboratory work-up. The short side chain sterols were found to possess predominantly the androst-5-en-3β-ol nucleus with C-17 alkyl side chains ranging from zero to six carbon atoms. Concentrations of short side chain sterols range from trace levels to over 5% of the sterol mixture in various species. The possible origins of these short side chain sterols are evaluated in the light of current knowledge of sterol function, biosynthesis, dealkylation, microbial degradation, and autoxidation. Known sterol autoxidations are reviewed, and possible singlet oxygen and free radical mechanisms of sterol side chain autoxidation (at physiological temperatures) which may lead to sterols with shortened hydrocarbon side chain are suggested. The possible autoxidative generation of short side chain sterols from known marine sterols by the suggested mechanisms is evaluated through application of the REACT computer program. Predicted short side chains are tabulated for each parent marine sterol side chain and then compared with the compositions of the actual sterols found in the marine extracts examined. The possible natural environmental or in vivo autoxidative formation of the short side chain marine sterols is supported by these evaluations.     

Conformational differences between sn-3-phospholipids and sn-2-phospholipids. A neutron and x-ray diffraction investigation

Extensive work has been reported on the conformation in membranes of sn-3-phosphatidylcholines, -ethanolamines, -glycerols and -serines (sn-3-phospholipids), where the headgroup is linked to the third carbon atom in the glycerol backbone. One important feature common to all these naturally occurring phospholipids is that the glycerol moiety is oriented almost perpendicular to the membrane surface, with the sn-1 chain continuing in this direction, whereas the sn-2 chain starts first in a direction parallel to the layer and then bends sharply at the second carbon atom. We present here neutron diffraction results on 1,3-dipalmitoyl-glycero-2-phosphocholine (1,3-DPPC) in which the headgroup is attached to the second carbon atom in the middle of the glycerol part and the two other adjacent carbon atoms are linked to the paraffin chains. Two 1,3-DPPC samples. ²H-labelled at different positions, were studied. One sample was deuterated at the first methylene segment in each fatty acyl chain, and the other at the first segment in one chain and at the second segment in the other chain. The Patterson maps as well as the neutron density profiles show that both fatty acyl chains in 1,3-DPPC have the same conformation introduced by this symmetric chemical bond pattern. It is concluded from this that the C₍₁₎C₍₃₎ vector of the glycerol backbone part is oriented parallel to the bilayer surface and the ²H nuclear magnetic resonance signals indicate that both chains have a conformation similar to that observed for the bent chain in sn-3-phospholipids. These findings indirectly confirm the idea that an intramolecular energy minimum, together with the packing geometry of the lipids in the membrane, produce the characteristic conformation around the glycerol backbone as is found for all naturally occurring phospholipids that have been studied so far.     

Relaxivities of paramagnetic liposomes: on the importance of the chain type and the length of the amphiphilic complex

Nuclear magnetic relaxation dispersion (NMRD) profiles of unilamellar DPPC liposomes incorporating Gd-DTPA-bisamides with alkyl chains of 12 to 18 C atoms in their external and internal layers were recorded in order to study the influence that the chain length and structure of Gd-bisamides incorporated in the liposomal membrane have on their proton relaxivity. The NMRD profiles recorded at 310 K show that the relaxivity reaches a minimum value when the carbon chain lengths of the phospholipid and of the Gd complex match and is at a maximum in the presence of a carbon-carbon double bond. For these DPPC paramagnetic liposomes, the longer the aliphatic chains of the complex, the larger will be its immobilization in the membrane. In addition, the presence of an unsaturated carbon-carbon bond in the alkyl chain of the Gd complex induces an increase of its mobility and of its water exchange rate with, as a result, a much greater efficiency as an MRI contrast agent.     

N-acylphosphatidylethanolamines: effect of the N-acyl chain length on its orientation

N-Acylphosphatidylethanolamines, or NAPEs, are found in tissues involved in degenerating processes, such as dehydrated endosperm of seeds, erythrocyte membranes, or cell injury. To determine the conformation and orientation of the acyl chains of these phospholipids, NAPEs with deuterated N-acyl chains of 6 and 16 carbon atoms were synthesized and studied by transmission and attenuated total reflectance (ATR) infrared spectroscopy. For N-C16d-DPPE, the ATR measurements show that the N-acyl chain has the same orientation as the two acyl chains attached to the glycerol moiety, while the N-acyl chain of N-C6d-DPPE is randomly oriented. These results demonstrate that for N-C16d-DPPE, the N-acyl chain is embedded into the hydrophobic core of the bilayer, while for the short chain derivative the N-acyl chain remains in the lipid headgroup region. The analysis of the carbonyl stretching band and of the amide I band suggests that, for the long N-acyl chain lipid, the ester C=O and the N-H groups are linked by intermolecular hydrogen bonds.     

Trifluoromethylated carboline compounds targeting DNA: Synthesis,binding and anti-proliferative effects on human cancer cell lines

Three sets of carboline derived compounds were prepared by Pictet-Spengler cyclization. These tetrahydro β- and γ-carbolines have CF₃ group with an additional amino alkyl chains (α- or δ-position) and guanidine alkyl chains (α-position), of varying length. Structure–activity relationship of these molecules with calf thymus DNA was emphasized by fluorescence, ITC, FTIR and viscosity. Binding with DNA resulted in dramatic enhancement and quenching in the fluorescence emission. Gamma-carboline analogs showed maximum DNA binding followed by beta-carboline compounds with amino alkyl chain and least with guanidine alkyl chain compounds. It decreased with increasing chain length. The bindings were entropically driven being more with guanidine alkyl chain analogs. Site preference and mode of binding with partial intercalation and external binding was supported by FTIR and viscosity. Cytotoxic potencies of the compounds were tested on seven different cancer cell lines. The smallest alkyl chain analog attached to gamma position, Comp3, showed maximum cytotoxicity with GI₅₀ 6.2 µM, against HCT-116 causing apoptosis, followed by the guanidine alkyl chain compounds, but amino alkyl chain compounds to beta position showed poor cytotoxicity.These results may be of prospective use in a framework to design novel carboline derivatives as antitumor drugs for improved therapeutic applications in future.     

Statistical and energetic analysis of side-chain conformations in oligopeptides

The distributions of side-chain conformations in 258 crystal structures of oligopeptides have been analyzed. The sample contains 321 residues having side chains that extend beyond the C beta atom. Statistically observed preferences of side-chain dihedral angles are summarized and correlated with stereochemical and energetic constraints. The distributions are compared with observed distributions in proteins of known X-ray structures and with computed minimum-energy conformations of amino acid derivatives. The distributions are similar in all three sets of data, and they appear to be governed primarily by intraresidue interactions. In side chains with no beta-branching, the most important interactions that determine chi 1 are those between the C gamma H2 group and atoms of the neighboring peptide groups. As a result, the g- conformation (chi 1 congruent to -60 degrees) occurs most frequently for rotation around the C alpha-C beta bond in oligopeptides, followed by the t conformation (chi 1 congruent to 180 degrees), while the g+ conformation (chi 1 congruent to 60 degrees) is least favored. In residues with beta-branching, steric repulsions between the C gamma H2 or C gamma H3 groups and backbone atoms govern the distribution of chi 1. The extended (t) conformation is highly favored for rotation around the C beta-C gamma and C gamma-C delta bonds in unbranched side chains, because the t conformer has a lower energy than the g+ and g- conformers in hydrocarbon chains. This study of the observed side-chain conformations has led to a refinement of one of the energy parameters used in empirical conformational energy computations.