RESUMEN
We report on neutron diffraction experiments performed on organogels prepared from triarylamine tris-amide (TATA), as well as on their ternary thermoreversible gels made up with poly[vinyl chloride] (PVC). Three different solvents together with their deuterated counterparts have been used; tetrachloroethane, wherein TATA fibrils display ohmic conductivity, bromobenzene and o-dichlorobenzene. The TATA crystal structure differs in the three solvents. Most importantly, the difference in the diffraction patterns whether hydrogenous solvents or deuterated solvents are used demonstrate the occurrence of molecular compounds. Tentative unit cells are presented. These results are also discussed in the light of the current views on the solvent role in the gelation process.
RESUMEN
Combined neutron scattering and diffusion nuclear magnetic resonance experiments have been used to reveal significant interregional asymmetries (lateralization) in bovine brain hemispheres in terms of myelin arrangement and water dynamics at micron to atomic scales. Thicker myelin sheaths were found in the left hemisphere using neutron diffraction. 4.7 T dMRI and quasi-elastic neutron experiments highlighted significant differences in the properties of water dynamics in the two hemispheres. The results were interpreted in terms of hemisphere-dependent cellular composition (number of neurons, cell distribution, etc.) as well as specificity of neurological functions (such as preferential networking).
Asunto(s)
Cerebelo/diagnóstico por imagen , Cerebro/diagnóstico por imagen , Imagen de Difusión por Resonancia Magnética/métodos , Dominancia Cerebral , Difusión Facilitada/fisiología , Difracción de Neutrones/métodos , Dispersión del Ángulo Pequeño , Agua/química , Animales , Bovinos , Vaina de Mielina/ultraestructuraRESUMEN
Water diffusion is an optimal tool for investigating the architecture of brain tissue on which modern medical diagnostic imaging techniques rely. However, intrinsic tissue heterogeneity causes systematic deviations from pure free-water diffusion behaviour. To date, numerous theoretical and empirical approaches have been proposed to explain the non-Gaussian profile of this process. The aim of this work is to shed light on the physics piloting water diffusion in brain tissue at the micrometre-to-atomic scale. Combined diffusion magnetic resonance imaging and first pioneering neutron scattering experiments on bovine brain tissue have been performed in order to probe diffusion distances up to macromolecular separation. The coexistence of free-like and confined water populations in brain tissue extracted from a bovine right hemisphere has been revealed at the micrometre and atomic scale. The results are relevant for improving the modelling of the physics driving intra- and extracellular water diffusion in brain, with evident benefit for the diffusion magnetic resonance imaging technique, nowadays widely used to diagnose, at the micrometre scale, brain diseases such as ischemia and tumours.
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Encéfalo/fisiología , Bovinos/fisiología , Imagen por Resonancia Magnética , Difracción de Neutrones , Agua/metabolismo , AnimalesRESUMEN
The extracellular lipid matrix in the skin's outermost layer, the stratum corneum, is crucial for the skin barrier. The matrix is composed of ceramides (CERs), cholesterol (CHOL) and free fatty acids (FFAs) and involves two lamellar phases: the short periodicity phase (SPP) and the long periodicity phase (LPP). To understand the skin barrier thoroughly, information about the molecular arrangement in the unit cell of these lamellar phases is paramount. Previously we examined the molecular arrangement in the unit cell of the SPP. Furthermore X-ray and neutron diffraction revealed a trilayer arrangement of lipids within the unit cell of the LPP [D. Groen et al., Biophysical Journal, 97, 2242-2249, 2009]. In the present study, we used neutron diffraction to obtain more details about the location of lipid (sub)classes in the unit cell of the LPP. The diffraction pattern revealed at least 8 diffraction orders of the LPP with a repeating unit of 129.6±0.5Å. To determine the location of lipid sub(classes) in the unit cell, samples were examined with either only protiated lipids or selectively deuterated lipids. The diffraction data obtained by means of D2O/H2O contrast variation together with a gradual replacement of one particular CER, the acyl CER, by its partly deuterated counterpart, were used to construct the scattering length density profiles. The acyl chain of the acyl CER subclass is located at a position of ~21.4±0.2Å from the unit cell centre of the LPP. The position and orientation of CHOL in the LPP unit cell were determined using tail and head-group deuterated forms of the sterol. CHOL is located with its head-group positioned ~26±0.2Å from the unit cell centre. This allows the formation of a hydrogen bond with the ester group of the acyl CER located in close proximity. Based on the positions of the deuterated moieties of the acyl CER, CHOL and the previously determined location of two other lipid subclasses [E.H. Mojumdar et al., Biophysical Journal, 108, 2670-2679, 2015], a molecular model is proposed for the unit cell of the LPP. In this model CHOL is located in the two outer layers of the LPP, while CER EOS is linking the two outer layers with the central lipid layers. Finally the two other lipid subclasses are predominantly located in the central layer of the LPP.
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Ceramidas/análisis , Colesterol/análisis , Epidermis/química , Agua Corporal , Óxido de Deuterio/análisis , Epidermis/ultraestructura , Ácidos Grasos no Esterificados/análisis , Ácidos Grasos no Esterificados/química , Ácido Linoleico/análisis , Lípidos/análisis , Lípidos/química , Estructura Molecular , Difracción de Neutrones , Absorción CutáneaRESUMEN
The intercellular lipid matrix of the skin's stratum corneum serves to protect the body against desiccation and simultaneously limits the passage of drugs and other xenobiotics into the body. The matrix is made up of ceramides, free fatty acids, and cholesterol, which are organized as two coexisting crystalline lamellar phases. In studies reported here, we sought to use the technique of neutron diffraction, together with the device of isotopic (H/D) substitution, to determine the molecular architecture of the lamellar phase having a repeat distance of 53.9 ± 0.3 Å. Using hydrogenous samples as well as samples incorporating perdeuterated (C24:0) fatty acids and selectively deuterated cholesterol, the diffraction data obtained were used to construct neutron scattering length density profiles. By this means, the locations within the unit cell were determined for the cholesterol and fatty acids. The cholesterol headgroup was found to lie slightly inward from the unit cell boundary and the tail of the molecule located 6.2 ± 0.2 Å from the unit cell center. The fatty acid headgroups were located at the unit cell boundary with their acyl chains straddling the unit cell center. Based on these results, a molecular model is proposed for the arrangement of the lipids within the unit cell.
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Membrana Celular/química , Membrana Celular/metabolismo , Colesterol/química , Colesterol/metabolismo , Ácidos Grasos/química , Ácidos Grasos/metabolismo , Difracción de Neutrones , Transporte Biológico , Ceramidas/química , Ceramidas/metabolismo , Células Epidérmicas , HumanosRESUMEN
In order to elucidate the mode of action of the lipophilic penetration enhancer isopropyl myristate (IPM) on a molecular scale, we investigated oriented quaternary stratum corneum (SC) lipid model membranes based on ceramide AP, cholesterol, palmitic acid and cholesterol sulfate containing 10 wt% IPM by means of neutron diffraction. Our results indicate that IPM affects the lamellar lipid assembly in terms of bilayer perturbation and disordering. Phase segregation occurred, indicating that IPM is not likely to mix properly with the other SC lipids due to its branched structure. We used selective deuterium labelling to localize the penetration enhancer, and could successfully prove the presence of IPM in the two coexisting lamellar phases. We conclude that IPM's mode of action as penetration promoter is presumably based on incorporation into the SC lipid matrix, extraction of certain SC lipids into a separate phase and perturbation of the multilamellar lipid assembly.
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Epidermis/efectos de los fármacos , Lípidos de la Membrana/metabolismo , Miristatos/farmacología , Absorción Cutánea/efectos de los fármacos , Deuterio , Epidermis/metabolismo , Membranas Artificiales , Difracción de Neutrones , PermeabilidadRESUMEN
The lipid matrix present in the uppermost layer of the skin, the stratum corneum, plays a crucial role in the skin barrier function. The lipids are organized into two lamellar phases. To gain more insight into the molecular organization of one of these lamellar phases, we performed neutron diffraction studies. In the diffraction pattern, five diffraction orders were observed attributed to a lamellar phase with a repeat distance of 5.4 nm. Using contrast variation, the scattering length density profile could be calculated showing a typical bilayer arrangement. To obtain information on the arrangement of ceramides in the unit cell, a mixture that included a partly deuterated ceramide was also examined. The scattering length density profile of the 5.4-nm phase containing this deuterated ceramide demonstrated a symmetric arrangement of the ceramides with interdigitating acyl chains in the center of the unit cell.
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Membrana Celular/química , Ceramidas/química , Difracción de Neutrones , Supervivencia Celular , Colesterol/química , Células Epidérmicas , Humanos , Agua/químicaRESUMEN
The conformation of a linear polymer chain is studied as a function of the concentration of a macromolecular crowding agent by neutron scattering. Excluded volume to random coil due to macromolecular crowding in cells is predicted to exert a compressive force that will tend to reduce its size. It is shown that when reducing free volume due to macromolecular crowding, we observe a compression of the polymer chain with a reduction in its radius of gyration of up to approximately 30% and that the effective chain-chain interactions are strongly modified.
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Conformación Molecular , Polímeros/química , Difracción de Neutrones , Rotación , Dispersión del Ángulo PequeñoRESUMEN
Spherical polyelectrolyte block copolymer micelles were investigated as a function of added NaCl salt concentration using Small-Angle Neutron Scattering (SANS) and Light Scattering (LS). The micelles are formed by the self-association of charged-neutral copolymers made of a long deuterated polyelectrolyte moiety (NaPSS(d))251 and a short hydrophobic moiety (PEP)52. In presence of salt, the core shape and the aggregation number of the micelles are not affected. The hydrodynamic radius of the micelle is found to be identical to the radius of the whole micelle deduced from neutron scattering and thus the hydrodynamic radius is a valid measure of the corona thickness. At the lowest salt concentrations investigated the thickness of the corona, R(s), remains essentially constant and a contraction is observed above an added-salt concentration c(s) of 2 x 10(-2) M where this crossover concentration corresponds to the average ionic strength of the free counterions in the corona. The contraction takes place while maintaining a rod-like behavior of the chains at short scale and obeys to: R(s) approximately c(s)(-0.18). The exponent 0.18 suggests an electrostatic persistence length proportional to the Debye screening length.
RESUMEN
Self-assembled structures having a regular hollow icosahedral form (such as those observed for proteins of virus capsids) can occur as a result of biomineralization processes, but are extremely rare in mineral crystallites. Compact icosahedra made from a boron oxide have been reported, but equivalent structures made of synthetic organic components such as surfactants have not hitherto been observed. It is, however, well known that lipids, as well as mixtures of anionic and cationic single chain surfactants, can readily form bilayers that can adopt a variety of distinct geometric forms: they can fold into soft vesicles or random bilayers (the so-called sponge phase) or form ordered stacks of flat or undulating membranes. Here we show that in salt-free mixtures of anionic and cationic surfactants, such bilayers can self-assemble into hollow aggregates with a regular icosahedral shape. These aggregates are stabilized by the presence of pores located at the vertices of the icosahedra. The resulting structures have a size of about one micrometre and mass of about 1010 daltons, making them larger than any known icosahedral protein assembly or virus capsid. We expect the combination of wall rigidity and holes at vertices of these icosahedral aggregates to be of practical value for controlled drug or DNA release.
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Soluciones , Tensoactivos/química , Aniones , Cationes , Cristalización , Glicerol/química , Estructura Molecular , Ácido Mirístico/química , Dispersión de RadiaciónRESUMEN
Discs of finite size are a very rare form of stable surfactant self-assembly. It is shown that mixing of two oppositely charged single-chain surfactants can produce rigid nanodiscs as well as swollen lamellar liquid crystals with frozen bilayers. The crucial requirement for obtaining nanodisc self-assembly is the use of H+ and OH- as counterions. These counterions then form water and lower the conductivity to 10 microsiemens per centimeter. In the case of cationic component excess, a dilute solution of nanodiscs is in thermodynamic equilibrium with a lamellar phase. The diameter of the cationic nanodiscs is continuously adjustable from a few micrometers to 30 nanometers, with the positive charge located mainly around the edges.