A lithography-free pathway for chemical microstructuring of macromolecules from aqueous solution based on wrinkling

We report on a novel lithography-free method for obtaining chemical submicron patterns of macromolecules on flat substrates. The approach is an advancement of the well-known microcontact printing scheme: While for classical microcontact printing lithographically produced masters are needed, we show that controlled wrinkling can serve as an alternative pathway to producing such masters. These can even show submicron periodicities. We expect upscaling to larger areas to be considerably simpler than that for existing techniques, as wrinkling results in a macroscopic deformation process that is not limited in terms of substrate size. Using this approach, we demonstrate successful printing of aqueous solutions of polyelectrolytes and proteins. We study the effectiveness of the stamping process and its limits in terms of periodicities and heights of the stamps' topographical features. We find that critical wavelengths are well below 355 nm and critical amplitudes are below 40 nm and clarify the failure mechanism in this regime. This will permit further optimization of the approach in the future.     

Epifluorescence imaging of electrochemically switchable Langmuir-Blodgett films of Nafion

A combination of electrochemistry and luminescence methods was exploited to obtain information on the electrochemical activity and homogeneity of Nafion Langmuir-Blodgett films. The redox behavior of the Ru(bpy)3(2+) probe incorporated in the Nafion film was monitored by epifluorescence microscopy. The photoluminescent images, recorded by a charge-coupled device (CCD) camera, reflect the distribution of the probe in the film, which resulted as very uniform, particularly in comparison with spin-coated films. Apparent diffusion coefficients (Dapp) determined by cyclic voltammetry for films of less than 10 layers are in the range of 1 x 10(-12) to 8 x 10(-12) cm(2) s(-1), that is, 2 orders of magnitude lower than values reported in the literature for spin-coated Nafion films. The application to the electrode of a potential able to oxidize the luminescent Ru(bpy)3(2+) to the nonluminescent Ru(bpy)3(3+) switched off the photoluminescence with a response time that for the LB films was much shorter than that for the spin-coated ones. Experimental evidence and calculations indicate that lowering of the film thickness down to the nanometric level is very effective in shortening the switching time, notwithstanding the lowering of the Dapp value in LB films.     

Tuning nanoscopic water layers on hydrophobic and hydrophilic surfaces with laser light

The evolutional function of ordered interfacial water near solid surfaces was postulated by Szent-Gy?rgyi: "Life actually, may have started with building these water structures." Here we report their tunability with laser light on both hydrophobic and hydrophilic surfaces. On the former, the light caused their depletion--on the latter, an increase in fluidity--as measured by atomic force acoustic microscopy. Interfacial water layers play a key role in cellular recognition. Their tunability promises to revolutionize various fields in biomedical engineering and life sciences.     

Granulating titania powder by colloidal route using polyelectrolytes

A new, convenient, and inexpensive approach to process and granulate titania powders by a chemical route is proposed. It is based on the use of a formulation that includes a polyanion such as poly(sodium 4-styrenesulfonate) (PSS). Such a polyelectrolyte is most often considered to achieve dispersion of oxide powders in water. Basically, it adsorbs onto the surface of particles and induces electrical and/or steric interactions between particles in the suspension, which prevents agglomeration and rapid sedimentation. The advantages of polyelectrolytes in ceramic processing is well documented in the literature to produce low viscosity suspensions that are further used to form ceramic parts. In the case of TiO2 powders, such aqueous dispersions were obtained by adding small quantities of PSS. However, when exploring the behavior of mixtures containing lower contents of dispersant, we have discovered that, well below the optimum concentration required to get stable dispersions, the polyelectrolyte can act as a binder for titania particles. This can confer cohesion to the agglomerates, which can be processed to form large size (e.g., millimeter size) spheres. This phenomenon takes place when the oxide surface carries both positive and negative electrical charges and can be explained on a simple basis involving surface chemistry. For the optimum concentration of PSS that disperses titania, a polycation such as chitosan should be added to get spheres. This simple technique is expected to receive increasing attention due its potentialities and strong advantages with respect to other granulation techniques, such as spray-drying, which are energy consuming.     

Total synthesis of discodermolide: optimization of the effective synthetic route

An efficient and modulable total synthesis of discodermolide (DDM), a unique marine anticancer polyketide is described including related alternative synthetic approaches. Particularly notable is the repeated application of a crotyltitanation reaction to yield homoallylic (Z)-O-ene-carbamate alcohols with excellent selectivity. Advantage was taken of this reaction not only for the stereocontrolled building of the syn-anti methyl-hydroxy-methyl triads of DDM, but also for the direct construction of the terminal (Z)-diene. Of particular interest is also the installation of the C13=C14 (Z)-double bond through a highly selective dyotropic rearrangement. The preparation of the middle C8-C14 fragment in two sequential stages and its coupling to the C1-C7 moiety was a real challenge and required careful optimization. Several synthetic routes were explored to allow high and reliable yields. Due to the flexibility and robust character of this approach, it might enable a systematic structural variation of DDM and, therefore, the elaboration and exploration of novel discodermolide structural analogues.     

Gas-liquid selective oxidations with oxygen under explosive conditions in a micro-structured reactor

The gas-liquid oxidation of cyclohexane is performed at high temperature (>200 degrees C) and pressure (up to 25 bar) using pure oxygen in a Pyrex capped silicon etched microreactor which allows convenient screen reaction conditions well above the flammability limit.     

N-benzyl aspartate nitrones: unprecedented single-step synthesis and [3 + 2] cycloaddition reactions with alkenes

N-benzyl aspartate nitrones 2, prepared by addition of N-benzylhydroxylamine to dialkyl acetylenedicarboxylates 1, underwent [3 + 2] thermal cycloaddition with a wide range of alkenes to afford isoxazolidines 4 bearing a polyfunctionalized quaternary center. Under these uncatalyzed conditions, the trans stereocontrol observed with vinyl ethers is higher than that obtained with all acyclic activated nitrones reported to date. The first asymmetric access to a type-4 pure adduct was achieved starting from the chiral aspartate nitrone derived from (S)-alpha-methylbenzylhydroxylamine.     

Oligonucleotide nanostructured surfaces: effect on Escherichia coli curli expression

Oligonucleotide model surfaces allowing independent variation of topography and chemical composition were designed to study the adhesion and biofilm growth of E.coli. Surfaces were produced by covalent binding of oligonucleotides and immobilization of nucleotide-based vesicles. Their properties were confirmed through a combination of fluorescence microscopy, XPS, ellipsometry, AFM and wettability studies at each step of the process. These surfaces were then used to study the response of three different strains of E.coli quantified in a static biofilm growth mode. This study led to convincing evidence that oligonucleotide-modified surfaces, independent of the topographical feature used in this study, enhanced curli expression without an increase in the number of adherent bacteria.     

Identification of new<Emphasis Type="Italic"> O</Emphasis>-GlcNAc modified proteins using a click-chemistry-based tagging

The O-linked β-N-acetylglucosamine (O-GlcNAc) modification is an abundant post-translational modification in eukaryotic cells. This dynamic glycosylation plays a fundamental role in the activity of many nuclear and cytoplasmic proteins and is associated with pathologies like type II diabetes, Alzheimer’s disease or some cancers. However the exact link between O-GlcNAc-modified proteins and their function in cells is largely undefined for most cases. Here we report a strategy based on the 1,3-dipolar cycloaddition, called click chemistry, between unnatural N-acetylglucosamine (GlcNAc) analogues (substituted with an azido or alkyne group) and the corresponding biotinylated probe to specifically detect, enrich and identify O-GlcNAc-modified proteins. This bio-orthogonal conjugation confirms that only azido analogue of GlcNAc is metabolized by the cell. Thanks to the biotin probe, affinity purification on streptavidin beads allowed us to identify 32 O-GlcNAc-azido-tagged proteins by LC-MS/MS analysis in an MCF-7 cellular model, 14 of which were previously unreported. This work illustrates the use of the click-chemistry-based strategy combined with a proteomic approach to get further insight into the pattern of O-GlcNAc-modified proteins and the biological significance of this post-translational modification. Figure Detection of biotinylated O-GlcNAz proteins in MCF-7 cells Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. Caroline Gurcel and Anne-Sophie Vercoutter-Edouart contributed equally to this work.