Membrane assembly and ion transport ability of a fluorinated nanopore

A novel 21-residue peptide incorporating six fluorinated amino acids was prepared. It was designed to fold into an amphiphilic alpha helical structure of nanoscale length with one hydrophobic face and one fluorinated face. The formation of a fluorous interface serves as the main vector for the formation of a superstructure in a bilayer membrane. Fluorescence assays showed this ion channel's ability to facilitate the translocation of alkali metal ions through a phospholipid membrane, with selectivity for sodium ions. Computational studies showed that a tetramer structure is the most probable and stable supramolecular assembly for the active ion channel structure. The results illustrate the possibility of exploiting multiple Fd-:M+ interactions for ion transport and using fluorous interfaces to create functional nanostructures

Measurement of the Lateral Diffusion of Dipalmitoylphosphatidylcholine Adsorbed on Silica Beads in the Absence and Presence of Melittin: A 31P Two-Dimensional Exchange Solid-State NMR Study

Abstract31P two-dimensional exchange solid-state NMR spectroscopy was used to measure the lateral diffusion, DL, in the fluid phase of dipalmitoylphosphatidylcholine (DPPC) in the presence and absence of melittin. The use of a spherical solid support with a radius of 320±20nm, on which lipids and peptides are adsorbed together, and a novel way of analyzing the two-dimensional exchange patterns afforded a narrow distribution of DL centered at a value of (8.8±0.5)×10−8cm2/s for the pure lipid system and a large distribution of DL spanning 1×10−8 to 10×10−8cm2/s for the lipids in the presence of melittin. In addition, the determination of DL for nonsupported DPPC multilamellar vesicles (MLVs) suggests that the support does not slow down the lipid diffusion and that the radii of the bilayers vary from 300 to 800nm. Finally, the DPPC-melittin complex is stabilized at the surface of the silica beads in the gel phase, opening the way to further study of the interaction between melittin and DPPC

Using infrared and raman microspectroscopies to compare ex vivo involved psoriatic skin with normal human skin

Psoriasis is a chronic dermatosis that affects around 3% of the world’s population. The etiology of this autoimmune pathology is not completely understood. The barrier function of psoriatic skin is known to be strongly altered, but the structural modifications at the origin of this dysfunction are not clear. To develop strategies to reduce symptoms of psoriasis or adequate substitutes for modeling, a deep understanding of the organization of psoriatic skin at a molecular level is required. Infrared and Raman microspectroscopies have been used to obtain direct molecular-level information on psoriatic and healthy human skin biopsies. From the intensities and positions of specific vibrational bands, the lipid and protein distribution and the lipid order have been mapped in the different layers of the skin. Results showed a similar distribution of lipids and collagen for normal and psoriatic human skin. However, psoriatic skin is characterized by heterogeneity in lipid/protein composition at the micrometer scale, a reduction in the definition of skin layer boundaries and a decrease in lipid chain order in the stratum corneum as compared to normal skin. A global decrease of the structural organization is exhibited in psoriatic skin that is compatible with an alteration of its barrier properties

Transdermal diffusion, spatial distribution and physical state of a potential anticancer drug in mouse skin as studied by diffusion and spectroscopic techniques

Background:Understanding the efficiency of a transdermal medical drug requires the characterization of its diffusion process, including its diffusion rate, pathways and physical state. Objective:The aim of this work is to develop a strategy to achieve this goal. Methods:FTIR spectroscopic imaging in conjunction with a Franz cell and HPLC measurements were used to examine the transdermal penetration of deuterated tert-butyl phenylchloroethylurea (tBCEU), a molecule with a potential anticancer action. tBCEU has been solubilized in an expedient solvent mixture and its diffusion in hairless mouse skin has been studied. Results:The results indicate that tBCEU diffuses across the skin for more than 10 hours with a rate comparable to selegiline, an officially-approved transdermal drug. IR image analyses reveal that after 10 hours, tBCEU penetrates skin and that its spatial distribution does not correlate with neither the distribution of lipids nor proteins. tBCEU accumulates in cluster domains but overall low concentrations are found in skin. FTIR spectroscopic imaging additionally reveals that tBCEU is in a crystalline form. Conclusions:The results suggest that tBCEU is conveyed through the skin without preferential pathway. FTIR spectroscopic imaging and transdermal diffusion measurements appear as complementary techniques to investigate drug diffusion in skin

Magnetic resonance imaging of human tissue-engineered adipose substitutes

Adipose tissue (AT) substitutes are being developed to answer the strong demand in reconstructive surgery. To facilitate the validation of their functional performance in vivo, and to avoid resorting to excessive number of animals, it is crucial at this stage to develop biomedical imaging methodologies, enabling the follow-up of reconstructed AT substitutes. Until now, biomedical imaging of AT substitutes has scarcely been reported in the literature. Therefore, the optimal parameters enabling good resolution, appropriate contrast, and graft delineation, as well as blood perfusion validation, must be studied and reported. In this study, human adipose substitutes produced from adipose-derived stem/stromal cells using the self-assembly approach of tissue engineering were implanted into athymic mice. The fate of the reconstructed AT substitutes implanted in vivo was successfully followed by magnetic resonance imaging (MRI), which is the imaging modality of choice for visualizing soft ATs. T1-weighted images allowed clear delineation of the grafts, followed by volume integration. The magnetic resonance (MR) signal of reconstructed AT was studied in vitro by proton nuclear magnetic resonance (1H-NMR). This confirmed the presence of a strong triglyceride peak of short longitudinal proton relaxation time (T1) values (200±53 ms) in reconstructed AT substitutes (total T1=813±76 ms), which establishes a clear signal difference between adjacent muscle, connective tissue, and native fat (total T1 ∼300 ms). Graft volume retention was followed up to 6 weeks after implantation, revealing a gradual resorption rate averaging at 44% of initial substitute's volume. In addition, vascular perfusion measured by dynamic contrast-enhanced-MRI confirmed the graft's vascularization postimplantation (14 and 21 days after grafting). Histological analysis of the grafted tissues revealed the persistence of numerous adipocytes without evidence of cysts or tissue necrosis. This study describes the in vivo grafting of human adipose substitutes devoid of exogenous matrix components, and for the first time, the optimal parameters necessary to achieve efficient MRI visualization of grafted tissue-engineered adipose substitutes

Crown ether modified peptide interactions with model membranes

A simple model of an uncharged antimicrobial peptide, carrying four crown ether side chains, is modified further by the selective incorporation of arginine side chains to control its secondary structure and its interaction with model membranes and living cells. Conformational studies show that shifting the position of a cationic residue in the peptide sequence allows to control its secondary structure and supramolecular self-assembly in solution. Results also demonstrate that the secondary structure influences the interaction with model membranes and cells. An α-helical peptide with greater amphiphilicity forms assemblies that interact with both prokaryotic and eukaryotic model membranes and cells. However, a β-stranded peptide with evenly distributed charges generates assemblies that interact more selectively with prokaryotic model membranes and cells. In addition, we observed differences in peptide orientation between uncharged and cationic α-helical peptides with different phospholipid bilayers. In general, the studied peptides have a higher affinity for thinner membranes, and cationic peptides interacted better with anionic membranes

Avant-propos

Auger Danièle, Rossellini Michèle, Saïd Suzanne. Avant-propos. In: Cahiers de Fontenay, n°17, 1979. Aristophane : Les Femmes et la Cité. p. 7

Major Ampullate Spider Silk with Indistinguishable Spidroin Dope Conformations Leads to Different Fiber Molecular Structures

To plentifully benefit from its properties (mechanical, optical, biological) and its potential to manufacture green materials, the structure of spider silk has to be known accurately. To this aim, the major ampullate (MA) silk of Araneus diadematus (AD) and Nephila clavipes (NC) has been compared quantitatively in the liquid and fiber states using Raman spectromicroscopy. The data show that the spidroin conformations of the two dopes are indistinguishable despite their specific amino acid composition. This result suggests that GlyGlyX and GlyProGlyXX amino acid motifs (X = Leu, Glu, Tyr, Ser, etc.) are conformationally equivalent due to the chain flexibility in the aqueous environment. Species-related sequence specificity is expressed more extensively in the fiber: the β-sheet content is lower and width of the orientation distribution of the carbonyl groups is broader for AD (29% and 58°, respectively) as compared to NC (37% and 51°, respectively). β-Sheet content values are close to the proportion of polyalanine segments, suggesting that β-sheet formation is mainly dictated by the spidroin sequence. The extent of molecular alignment seems to be related to the presence of proline (Pro) that may decrease conformational flexibility and inhibit chain extension and alignment upon drawing. It appears that besides the presence of Pro, secondary structure and molecular orientation contribute to the different mechanical properties of MA threads