A modular spring-loaded actuator for mechanical activation of membrane proteins.

How cells respond to mechanical forces by converting them into biological signals underlie crucial cellular processes. Our understanding of mechanotransduction has been hindered by technical barriers, including limitations in our ability to effectively apply low range piconewton forces to specific mechanoreceptors on cell membranes without laborious and repetitive trials. To overcome these challenges we introduce the Nano-winch, a robust, easily assembled, programmable DNA origami-based molecular actuator. The Nano-winch is designed to manipulate multiple mechanoreceptors in parallel by exerting fine-tuned, low- piconewton forces in autonomous and remotely activated modes via adjustable single- and double-stranded DNA linkages, respectively. Nano-winches in autonomous mode can land and operate on the cell surface. Targeting the device to integrin stimulated detectable downstream phosphorylation of focal adhesion kinase, an indication that Nano-winches can be applied to study cellular mechanical processes. Remote activation mode allowed finer extension control and greater force exertion. We united remotely activated Nano-winches with single-channel bilayer experiments to directly observe the opening of a channel by mechanical force in the force responsive gated channel protein, BtuB. This customizable origami provides an instrument-free approach that can be applied to control and explore a diversity of mechanotransduction circuits on living cells.

In situ observation of the percolation threshold in multiphase magma analogues.

Magmas vesiculate during ascent, producing complex interconnected pore networks, which can act as outgassing pathways and then deflate or compact to volcanic plugs. Similarly, in-conduit fragmentation events during dome-forming eruptions create open systems transiently, before welding causes pore sealing. The percolation threshold is the first-order transition between closed- and open-system degassing dynamics. Here, we use time-resolved, synchrotron-source X-ray tomography to image synthetic magmas that go through cycles of opening and closing, to constrain the percolation threshold Φ C at a range of melt crystallinity, viscosity and overpressure pertinent to shallow magma ascent. During vesiculation, we observed different percolative regimes for the same initial bulk crystallinity depending on melt viscosity and gas overpressure. At high viscosity (> 106 Pa s) and high overpressure (~ 1-4 MPa), we found that a brittle-viscous regime dominates in which brittle rupture allows system-spanning coalescence at a low percolation threshold (Φ C ~0.17) via the formation of fracture-like bubble chains. Percolation was followed by outgassing and bubble collapse causing densification and isolation of the bubble network, resulting in a hysteresis in the evolution of connectivity with porosity. At low melt viscosity and overpressure, we observed a viscous regime with much higher percolation threshold (Φ C > 0.37) due to spherical bubble growth and lower degree of crystal connection. Finally, our results also show that sintering of crystal-free and crystal-bearing magma analogues is characterised by low percolation thresholds (Φ C = 0.04 - 0.10). We conclude that the presence of crystals lowers the percolation threshold during vesiculation and may promote outgassing in shallow, crystal-rich magma at initial stages of Vulcanian and Strombolian eruptions.

Development of specific dengue virus 2'-O- and N7-methyltransferase assays for antiviral drug screening.

Dengue virus (DENV) protein NS5 carries two mRNA cap methyltransferase (MTase) activities involved in the synthesis of a cap structure, (7Me)GpppA(2'OMe)-RNA, at the 5'-end of the viral mRNA. The methylation of the cap guanine at its N7-position (N7-MTase, (7Me)GpppA-RNA) is essential for viral replication. The development of high throughput methods to identify specific inhibitors of N7-MTase is hampered by technical limitations in the large scale synthesis of long capped RNAs. In this work, we describe an efficient method to generate such capped RNA, GpppA(2'OMe)-RNA74, by ligation of two RNA fragments. Then, we use GpppA(2'OMe)-RNA74 as a substrate to assess DENV N7-MTase activity and to develop a robust and specific activity assay. We applied the same ligation procedure to generate (7Me)GpppA-RNA74 in order to characterize the DENV 2'-O-MTase activity specifically on long capped RNA. We next compared the N7- and 2'-O-MTase inhibition effect of 18 molecules, previously proposed to affect MTase activities. These experiments allow the validation of a rapid and sensitive method easily adaptable for high-throughput inhibitor screening in anti-flaviviral drug development.

Phase-controlling phononic crystals: realization of acoustic Boolean logic gates.

A phononic crystal (PC) consisting of a square array of cylindrical polyvinylchloride inclusions in air is used to construct a variety of acoustic logic gates. In a certain range of operating frequencies, the PC band structure shows square-like equi-frequency contours centered off the gamma point. This attribute allows for the realization of non-collinear wave and group velocity vectors in the PC wave vector space. This feature can be utilized to control with great precision, the relative phase between propagating acoustic waves in the PC. By altering the incidence angle of the impinging acoustic beams or varying the PC thickness, interferences occur between acoustic wave pairs. It is recognized that information can be encoded with this mechanism (e.g., wave amplitudes/interference patterns) and accordingly to construct a series of logic gates emulating Boolean functions. The NAND, XOR, and NOT gates are demonstrated with finite-difference time-domain simulations of acoustic waves impinging upon the PC.

Simultaneous existence of phononic and photonic band gaps in periodic crystal slabs.

We discuss the simultaneous existence of phononic and photonic band gaps in a periodic array of holes drilled in a Si membrane. We investigate in detail both the centered square lattice and the boron nitride (BN) lattice with two atoms per unit cell which include the simple square, triangular and honeycomb lattices as particular cases. We show that complete phononic and photonic band gaps can be obtained from the honeycomb lattice as well as BN lattices close to honeycomb. Otherwise, all investigated structures present the possibility of a complete phononic gap together with a photonic band gap of a given symmetry, odd or even, depending on the geometrical parameters.

Experimental and theoretical evidence for subwavelength imaging in phononic crystals.

We show experimentally and theoretically that super resolution can be achieved while imaging with a flat lens consisting of a phononic crystal exhibiting negative refraction. This phenomenon is related to the coupling between the incident evanescent waves and a bound slab mode of the phononic crystal lens, leading to amplification of evanescent waves by the slab mode. Super resolution is only observed when the source is located very near to the lens, and is very sensitive to the location of the source parallel to the lens surface as well as to site disorder in the phononic crystal lattice.

Glycosylation reaction via a mild and efficient one-pot reaction using doped natural phosphate with iodine as catalyst.

Several alpha/beta-D-ribonucleosides were synthesized in good yields under mild conditions by N-glycosylations of acetyl 2,3,5-tri-O-benzoyl-beta-D-ribofuranose with silylated nucleobases in acetonitrile using NP doped with iodine as catalyst.

A glutaric acid ester as carrier system for sustained delivery of lamuvidine (3TC) dimers.

We report the synthesis of homo and heterodimers of 3TC conjugates. All new dimers were screened for their ability to inhibit HIV-1 in MT4 cell line and were compared to AZT alone and showed marked antiviral activity.

Multicoaxial cylindrical inclusions in locally resonant phononic crystals.

It is known that the transmission spectrum of the so-called locally resonant phononic crystal can exhibit absolute sharp dips in the sonic frequency range due to the resonance scattering of elastic waves. In this paper, we study theoretically, using a finite difference time domain method, the propagation of acoustic waves through a two-dimensional locally resonant crystal in which the matrix is a fluid (such as water) instead of being a solid as in most of the previous papers. The transmission is shown to be dependent upon the fluid or solid nature of the matrix as well as upon the nature of the coating material in contact with the matrix. The other main purpose of this paper is to consider inclusions constituted by coaxial cylindrical multilayers consisting of several alternate shells of a soft material (such as a soft rubber) and a hard material (such as steel). With respect to the usual case of a hard core coated with a soft rubber, the transmission spectrum can exhibit in the same frequency range several peaks instead of one. If two or more phononic crystals are associated together, we find that the structure displays all the zeros of transmission resulting from each individual crystal. Moreover, we show that it is possible to overlap the dips by an appropriate combination of phononic crystals and create a larger acoustic stop band.

High-yield production of short GpppA- and 7MeGpppA-capped RNAs and HPLC-monitoring of methyltransfer reactions at the guanine-N7 and adenosine-2'O positions.

Many eukaryotic and viral mRNAs, in which the first transcribed nucleotide is an adenosine, are decorated with a cap-1 structure, (7Me)G5'-ppp5'-A(2'OMe). The positive-sense RNA genomes of flaviviruses (Dengue, West Nile virus) for example show strict conservation of the adenosine. We set out to produce GpppA- and (7Me)GpppA-capped RNA oligonucleotides for non-radioactive mRNA cap methyltransferase assays and, in perspective, for studies of enzyme specificity in relation to substrate length as well as for co-crystallization studies. This study reports the use of a bacteriophage T7 DNA primase fragment to synthesize GpppAC(n) and (7Me)GpppAC(n) (1 < or = n < or = 9) in a one-step enzymatic reaction, followed by direct on-line cleaning HPLC purification. Optimization studies show that yields could be modulated by DNA template, enzyme and substrate concentration adjustments and longer reaction times. Large-scale synthesis rendered pure (in average 99%) products (1 < or = n < or = 7) in quantities of up to 100 nmol starting from 200 nmol cap analog. The capped RNA oligonucleotides were efficient substrates of Dengue virus (nucleoside-2'-O-)-methyltransferase, and human (guanine-N7)-methyltransferase. Methyltransfer reactions were monitored by a non-radioactive, quantitative HPLC assay. Additionally, the produced capped RNAs may serve in biochemical, inhibition and structural studies involving a variety of eukaryotic and viral methyltransferases and guanylyltransferases.

Propagation and localization of electromagnetic waves in quasiperiodic serial loop structures.

We study the propagation of electromagnetic waves in one-dimensional quasiperiodic photonic band gap structures made of serial loop structures separated by segments. Different quasiperiodic structures such as Fibonacci, Thue-Morse, Rudin-Shapiro, and double period are investigated with special focus on the Fibonacci structure. Depending on the lengths of the two arms constituting the loops, one can distinguish two particular cases. (i) There are symmetric loop structures, which are shown to be equivalent to impedance-modulated mediums. In this case, it is found that besides the existence of extended and forbidden modes, some narrow frequency bands appear as defect modes in the transmission spectrum inside the gaps. These modes are shown to be localized within only one of the two types of blocks constituting the structure. An analysis of the transmission phase time enables us to derive the group velocity as well as the density of states in these structures. In particular, the stop bands (localized modes) may give rise to unusual (strong normal) dispersion in the gaps, yielding fast (slow) group velocities above (below) the velocity of light. (ii) There are also asymmetric loop structures, where the loops play the role of resonators that may introduce transmission zeros and hence additional gaps unnoticed in the case of simple impedance-modulated mediums. A comparison of the transmission amplitude and phase time of Fibonacci systems with those of other quasiperiodic systems is also outlined. In particular, it was shown that these structures present similar behaviors in the transmission spectra inside the regions of extended modes, whereas they present different localized modes inside the gaps. Experiments and numerical calculations are in very good agreement.

Homo and heterodimers of ddI, d4T and AZT: influence of (5'-5') thiolcabonate-carbamate linkage on anti-HIV activity.

New homo and heterodimers of ddI, d4T and AZT with (5-5) thiolcarbonate-carbamate linkages have been prepared with the aim of testing them against wild type and NNRTI resistant HIV mutants. The prepared dimers showed a low activity in comparison to the parent drug.

Natural phosphate doped with KI in HMDS: a mild and efficient reagent for alkylation and glycosylation of nucleobases.

Several D-ribonucleosides are prepared from 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranoside and trimethylsilylated nucleobases under mild conditions by using natural phosphate doped with KI as catalyst.

Chemoenzymatic syntheses of homo- and heterodimers of AZT and d4T, and evaluation of their anti-HIV activity.

Homo- and heterodimers of AZT and d4T, possessing carbonate and carbamate linkers, have been synthesized with the aim to enhance the antiviral activity of their components. Homo- and heterodimer carbamates showed weak anti-HIV activity. On the other hand, dinucleoside carbonates showed marked antiviral activity.

Tunable filtering and demultiplexing in phononic crystals with hollow cylinders.

Acoustic band gap (ABG) materials constituted of steel hollow cylinders immersed in water can exhibit a tunable narrow pass band (NPB) located inside their gap. We theoretically investigate, using the finite difference time domain (FDTD) method, the properties of waveguides composed of a row of hollow cylinders in a two-dimensional (2D) phononic crystal made of filled steel cylinders. These waveguides exhibit NPB's at frequencies slightly higher than their infinite periodic ABG counterpart. The frequency of the waveguide's NPB can be selected by adjusting the inner radius of the hollow cylinders or by changing the nature of the fluid that fills them. We show that a waveguide constituted of a row of hollow cylinders with different inner radii can transport waves at two different frequencies. By selectively filling the cylinders with water or mercury we have created an active device that permits the transmission of waves at one, both, or neither of these frequencies. Finally, we examine the multiplexing and demultiplexing capabilities of Y shaped waveguides constituted of hollow cylinders.

Simple nanometric plasmon multiplexer.

We present a simple multiplexing structure made of two discrete plasmon wires coupled by two metal nanoclusters. We show that this simple nanosystem can transfer one plasmon wavelength from one wire to the other. Closed-form relations between the transmission coefficients and the nanocluster distances are given to optimize the desired directional plasmon ejection.

[Acoustic hearing aid with an integrated noise generator in hearing-impaired subjects with tinnitus]. / Prothèse acoustique et bruiteur conjoint pour des sujets hypoacousiques et acouphéniques.

INTRODUCTION: Sound therapy has been studied in a group of 30 patients experiencing hearing loss and tinnitus. MATERIAL AND METHODS: Patients wore a BTE hearing-aid (TCI-COMBI by Siemens), which, in addition to acoustic amplification, can generate a broadband noise. Comparison of self-questionnaires completed by the patients during a 12 week period with amplification alone to a 12 week period with amplification and noise. RESULTS AND CONCLUSIONS: His study demonstrates that the level of suffering is significantly reduced during the period with added noise.

Synthesis of new homo and heterodimers of 2',3'-dideoxyinosine (ddI) using ester linkages.

A series of new homo and heterodimers of ddI has been synthesized. A glutarate diester spacer was used to covalently couple ddI onto ddI, AZT or d4T.

Synthesis and biological evaluation of some acyclic 4,6-disubstituted 1H-pyrazolo[3,4-d]pyrimidine nucleosides.

The chemical synthesis and biological evaluation of some acyclic alpha-[6-(1'-carbamoylalkylthio)-1H-pyrazolo[3,4-d]pyrimidin-4-yl]thioalkylamide nucleosides are described.

Imidazolethyl-phosphoramidate alpha-oligonucleotides.

Alpha-ODNs conjugated to imidazole groups via phosphoramidate internucleosidic linkages were synthesized. The presence of the imidazolethyl-phosphoramidate linkage improved the affinity of alpha-ODNs for their nucleic acid targets.