Dielectrophoresis-Driven Spreading of Immersed Liquid Droplets

In recent years electrowetting-on-dielectric (EWOD) has become an effective tool to control partial wetting. EWOD uses the liquid−solid interface as part of a capacitive structure that allows capacitive and interfacial energies to adjust by changes in wetting when the liquid−solid interface is charged due to an applied voltage. An important aspect of EWOD has been its applications in micro fluidics in chemistry and biology and in optical devices and displays in physics and engineering. Many of these rely on the use of a liquid droplet immersed in a second liquid due to the need either for neutral buoyancy to overcome gravity and shield against impact shocks or to encapsulate the droplet for other reasons, such as in microfluidic-based DNA analyses. Recently, it has been shown that nonwetting oleophobic surfaces can be forcibly wetted by nonconducting oils using nonuniform electric fields and an interface-localized form of liquid dielectrophoresis (dielectrowetting). Here we show that this effect can be used to create films of oil immersed in a second immiscible fluid of lower permittivity. We predict that the square of the thickness of the film should obey a simple law dependent on the square of the applied voltage and with strength dependent on the ratio of difference in permittivity to the liquid-fluid interfacial tension, Δε/γLF. This relationship is experimentally confirmed for 11 liquid−air and liquid−liquid combinations with Δε/γLF having a span of more than two orders of magnitude. We therefore provide fundamental understanding of dielectrowetting for liquid-in-liquid systems and also open up a new method to determine liquid−liquid interfacial tensions

CyBase: a database of cyclic protein sequences and structures, with applications in protein discovery and engineering

CyBase was originally developed as a database for backbone-cyclized proteins, providing search and display capabilities for sequence, structure and function data. Cyclic proteins are interesting because, compared to conventional proteins, they have increased stability and enhanced binding affinity and therefore can potentially be developed as protein drugs. The new CyBase release features a redesigned interface and internal architecture to improve user-interactivity, collates double the amount of data compared to the initial release, and hosts a novel suite of tools that are useful for the visualization, characterization and engineering of cyclic proteins. These tools comprise sequence/structure 2D representations, a summary of grafting and mutation studies of synthetic analogues, a study of N- to C-terminal distances in known protein structures and a structural modelling tool to predict the best linker length to cyclize a protein. These updates are useful because they have the potential to help accelerate the discovery of naturally occurring cyclic proteins and the engineering of cyclic protein drugs. The new release of CyBase is available at http://research1t.imb.uq.edu.au/cybas

X-ray Crystallographic Structure of an Artificial β-Sheet Dimer

This paper describes the X-ray crystallographic structure of a designed cyclic beta-sheet peptide that forms a well-defined hydrogen-bonded dimer that mimics beta-sheet dimers formed by proteins. The 54-membered ring macrocyclic peptide (1a) contains molecular template and turn units that induce beta-sheet structure in a heptapeptide strand that forms the dimerization interface. The X-ray crystallographic structure reveals the structures of the two "Hao" amino acids that help template the beta-sheet structure and the two delta-linked ornithine turn units that link the Hao-containing template to the heptapeptide beta-strand. The Hao amino acids adopt a conformation that resembles a tripeptide in a beta-strand conformation, with one edge of the Hao unit presenting an alternating array of hydrogen-bond donor and acceptor groups in the same pattern as that of a tripeptide beta-strand. The delta-linked ornithines adopt a conformation that resembles a hydrogen-bonded beta-turn, in which the ornithine takes the place of the i+1 and i+2 residues. The dimers formed by macrocyclic beta-sheet 1a resemble the dimers of many proteins, such as defensin HNP-3, the lambda-Cro repressor, interleukin 8, and the ribonuclease H domain of HIV-1 reverse transcriptase. The dimers of 1a self-assemble in the solid state into a barrel-shaped trimer of dimers in which the three dimers are arranged in a triangular fashion. Molecular modeling in which one of the three dimers is removed and the remaining two dimers are aligned face-to-face provides a model of the dimers of dimers of closely related macrocyclic beta-sheet peptides that were observed in solution

Three-dimensional structure of RTD-1, a cyclic antimicrobial defensin from rhesus macaque leukocytes

Most mammalian defensins are cationic peptides of 29-42 amino acids long, stabilized by three disulfide bonds. However, recently Tang et al. (1999, Science 286, 498-502) reported the isolation of a new defensin type found in the leukocytes of rhesus macaques. In contrast to all the other defensins found so far, rhesus theta defensin-1 (RTD-1) is composed of just 18 amino acids with the backbone cyclized through peptide bonds. Antibacterial activities of both the native cyclic peptide and a linear form were examined, showing that the cyclic form was 3-fold more active than the open chain analogue [Tang et al. (1999) Science 286, 498-502]. To elucidate the three-dimensional structure of RTD-1 and its open chain analogue, both peptides were synthesized using solid-phase peptide synthesis and tert-butyloxycarbonyl chemistry. The structures of both peptides in aqueous solution were determined from two-dimensional H-1 NMR data recorded at 500 and 750 MHz. Structural constraints consisting of interproton distances and dihedral angles were used as input for simulated-annealing calculations and water refinement with the program CNS. RTD-1 and its open chain analogue oRTD-1 adopt very similar structures in water. Both comprise an extended beta -hairpin structure with turns at one or both ends. The turns are well defined within themselves and seem to be flexible with respect to the extended regions of the molecules. Although the two strands of the beta -sheet are connected by three disulfide bonds, this region displays a degree of flexibility. The structural similarity of RTD-1 and its open chain analogue oRTD-1, as well as their comparable degree of flexibility, support the theory that the additional charges at the termini of the open chain analogue rather than overall differences in structure or flexibility are the cause for oRTD-1's lower antimicrobial activity. In contrast to numerous other antimicrobial peptides, RTD-1 does not display any amphiphilic character, even though surface models of RTD-1 exhibit a certain clustering of positive charges. Some amide protons of RTD-1 that should be solvent-exposed in monomeric beta -sheet structures show low-temperature coefficients, suggesting the possible presence of weak intermolecular hydrogen bonds

Interaction of RTD-1, a cyclic antimicrobial defensin from Rhesus macaque leukocytes, and its open chain analogue with membrane mimics

In contrast to other mammalian defensins, rhesus theta defensin-1 (RTD-1) is composed of just 18 amino acids with the backbone cyclized through peptide bonds. Antibacterial activities of both the native cyclic peptide and a linear form were examined, showing that the cyclic form was 3-fold more active than the open chain analogue, oRTD-1, although both peptides adopt very similar structures in water. It was suggested that the additional charges at the termini of oRTD-1 are the cause for its lower antimicrobial activity. Therefore, we studied the interaction of both peptides with membrane mimics composed of zwitterionic (PC) and negatively charged (PG) phospholipids, major lipid components of erythrocyte and bacterial cell membranes, respectively. Microcalorimetry showed that RTD-1 and oRTD-1 did not affect the phase behavior of PC liposomes, while in PG liposomes both peptides induced new phase transitions above the chain melting transition of the lipid. The shape and fraction differed between both peptides, depending also on their concentration, which will be discussed in terms of their antimicrobial activity

Diagnostic de fertilité du sol au Centre-Nord de la Côte d’Ivoire

La mise en valeur actuelle des sols, aussi bien par les petits paysans que par les grands fermiers, fait peu appel a l'utilisation d'engrais minéraux. La pratique de la jachère est très rare et les espèces et variétés ont parfois des places précises sur l'exploitation. Afin d'appréhender la logique de fertilité des sols chez le paysan, nous avons réalisé un diagnostic de sol en culture d'igname associée au mode de production. Les résultats montrent que Le niveau d'exploitation des sols de la zone a été modérément élevé (33 % < IR < 67 %). Dans ce système, la jachère est faiblement pratiquée dans les zones de Dabakala et Katiola, alors qu'à Bouaké et Béoumi, la pratique est chose courante. Le choix de la parcelle par le paysan est principalement guidé par la teneur en matière organique du sol. Les données chimiques obtenues confirment, en partie les observations des paysans, quant au statut organique et le niveau de fertilité du sol. La teneur en carbone du sol permet de distinguer deux groupes de sol bien que la différence ne soit pas significative (Duncan, p > F = 0,1236 ; α = 0,05). La partie sud de la zone (Sakassou et Béoumi) s'est distinguée des zones de Katiola, Bouaké et Dabakala.Today land use by both small and big farmers do not make use mineral fertilizer. Fallow practice is rare, with plant species often occupying a specific place in the field. In order to determine farmer perception for soil fertility, we conducted a diagnostic survey of soil under yam production. Results show that the level of soil exploitation in the area was moderately (33 % < IR < 67 %). In this system, fallow techniques are weakly practiced in Dabakala and Katiola zones, whereas in Bouaké and Béoumi, the practice was more frequent. Organic matter content was found to be the driving force in the choice of his yam plot by the farmer. The chemical data confirmed, in part, farmer observations, as for the organic status and the level of soil fertility. Two groups of soils, based on carbon contents were distinguished, although differences were not significant (P > F= 0.1236, α = 0.05 ; Duncan multiple range test). South of the area (Sakassou and Béoumi) were different from Katiola, Bouaké and Dabakala areas.Keywords: Diagnose, fertility, yam, organic matter, fallow, Côte d'Ivoir

Manipulated wettability of a superhydrophobic quartz crystal microbalance through electrowetting

The liquid phase response of quartz crystal microbalances (QCM) with a thin coating (~9 µm) of epoxy resin with and without a carbon nanoparticles top layer is reported. The nanoparticles convert the epoxy surface to a superhydrophobic one with a high static contact angle (~151º-155º) and low contact angle hysteresis (~1º-3.7º) where droplets of water are in the suspended Cassie-Baxter state. The frequency decrease of the fully immersed QCM with the superhydrophobic surface is less than with only epoxy layer, thus indicating a decoupling of the QCM response. A wettability transition to a liquid penetrating into the surface roughness state (for droplets a high contact angle hysteresis Wenzel state) was triggered using a molarity of ethanol droplet test (MED) and electrowetting; the MED approach caused some surface damage. The electrowetting induced transition caused a frequency decrease of 739 Hz at a critical voltage of ~100 V compared to the QCM in air. This critical voltage correlates to a contact angle decrease of 26º and a high contact angle hysteresis state in droplet experiments. These experiments provide a proof-of-concept that QCMs can be used to sense wetting state transitions and not only mass attachments or changes in viscosity-density products of liquids

The cyclic antimicrobial peptide RTD-1 induces stabilized lipid-peptide domains more efficiently than its open-chain analogue

The effects of a mammalian cyclic antimicrobial peptide, rhesus theta defensin 1 (RTD-1) and its open chain analogue (oRTD-1), on the phase behaviour and structure of model membrane systems (dipalmitoyl phosphatidylcholine, DPPC and dipalmitoyl phosphatidylglycerol, DPPG) were studied. The increased selectivity of RTD-1 for anionic DPPG over zwitterionic DPPC was shown by differential scanning calorimetry. RTD-1, at a molar peptide-lipid ratio of 1:100, induced considerable changes in the phase behaviour of DPPG, but not of DPPC. The main transition temperature, T-m, Was unchanged, but additional phase transitions appeared above T-m. oRTD-1 induced similar effects. However, the effects were not observable below a peptide:lipid molar ratio of 1:50, which correlates with the weaker biological activity of oRTD-1. Small-and wide-angle X-ray scattering revealed for DPPG the appearance of additional structural features induced by RTP-1 above T-m, which were interpreted as correlated lamellar structures, with increased order of the fatty acyl side chains of the lipid. It is proposed that after initial electrostatic interaction of the cationic rim of the peptide with the anionic DPPG headgroups, leading to stabilized lipid-peptide clusters, the hydrophobic face of the peptide assists in its interaction with the fatty acyl side chains eventually leading to membrane disruption. (C) 2004 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved