Tet-on, or tet-off, that is the question: advanced conditional gene expression in Aspergillus

In Aspergillus, controlled gene expression is often achieved using the reverse tetracycline-controlled transactivator (rtTA) dependent Tet-on system, whereby transcription is activated in a titratable manner by addition of the tetracycline derivative doxycycline. The complementary Tet-off system utilises the tetracycline-controlled transactivator (tTA) component to quantitatively reduce gene expression. In this study, we utilised a synthetic biological approach to engineer highly optimised Tet-off conditional expression systems in Aspergillus niger and Aspergillus fumigatus. Steps for delivery of these tools include utilising codon optimised cassette components, testing several promoters for improved genetic stability and validating two modified luciferase reporters for highly accurate measurements of gene expression. The Tet-off cassettes developed in this study enable facile and quantitative functional analysis, as validated by Tet-off analysis of genes involved in chitin synthesis and cell wall polarity in A. niger, and para-aminobenzoic acid synthesis in A. fumigatus. We also used a racAG18V dominant allele to demonstrate that Tet-off in A. niger enables gene over-expression and downregulation in a single isolate. Additionally, we used the improved luciferase reporters to show that the Tet-off cassette in A. niger enables quantification of gene oscillations. In order to demonstrate that synthetic biological approaches developed here are broadly applicable to engineering transcriptional circuits in filamentous fungi, we used our strategy for improving cassette stability by promoter replacement in the A. niger Tet-on system, which resulted in a modified Tet-on cassette with higher stability in recipient genomes

Fermiology and superconductivity studies on the non-tetrachalcogenafulvalene structured organic superconductor beta-(BDA-TTP)_2SbF_6

The quantum oscillatory effect and superconductivity in a non-tetrachalcogenafulvalene (TCF) structure based organic superconductor beta-(BDA-TTP)_2SbF_6 are studied. Here the Shubnikov-de Haas effect (SdH) and angular dependent magnetoresistance oscillations (AMRO) are observed. The oscillation frequency associated with a cylindrical Fermi surface is found to be about 4050 tesla, which is also verified by the tunnel diode oscillator (TDO) measurement. The upper critical field Hc2 measurement in a tilted magnetic field and the TDO measurement in the mixed state reveal a highly anisotropic superconducting nature in this material. We compared physical properties of beta-(BDA-TTP)_2SbF_6 with typical TCF structure based quasi two-dimensional organic conductors. A notable feature of beta-(BDA-TTP)_2SbF_6 superconductor is a large value of effective cyclotron mass m_c^*=12.4+/1.1 m_e, which is the largest yet found in an organic superconductor. A possible origin of the enhanced effective mass and its relation to the superconductivity are briefly discussed.Comment: 8 pages, 10 figure

Coherent vs incoherent interlayer transport in layered metals

The magnetic-field, temperature, and angular dependence of the interlayer magnetoresistance of two different quasi-two-dimensional (2D) organic superconductors is reported. For $\kappa$-(BEDT-TTF)$_2$I$_3$ we find a well-resolved peak in the angle-dependent magnetoresistance at $\Theta = 90^\circ$ (field parallel to the layers). This clear-cut proof for the coherent nature of the interlayer transport is absent for $\beta$''-(BEDT-TTF)$_2$SF$_5$CH$_2$CF$_2$SO$_3$. This and the non-metallic behavior of the magnetoresistance suggest an incoherent quasiparticle motion for the latter 2D metal.Comment: 4 pages, 4 figures. Phys. Rev. B, in pres

Interplane Transport and Superfluid Density in Layered Superconductors

We report on generic trends in the behavior of the interlayer penetration depth $\lambda_c$ of several different classes of quasi two-dimensional superconductors including cuprates, Sr$_2$RuO$_4$, transition metal dichalcogenides and organic materials of the $(BEDT-TTF)_2X$-series. Analysis of these trends reveals two distinct patterns in the scaling between the values of $\lambda_c$ and the magnitude of the DC conductivity: one realized in the systems with a Fermi liquid (FL) ground state and the other seen in systems with a marked deviation from the FL response. The latter pattern is found primarily in under-doped cuprates and indicates a dramatic enhancement (factor $\simeq 10^2$) of the energy scale $\Omega_C$ associated with the formation of the condensate compared to the data for the FL materials. We discuss implications of these results for the understanding of pairing in high-$T_c$ cuprates.Comment: 4 pages, 2 figure

Drop test: A new method to measure the particle adhesion force

Measurement of the adhesive force is of great interest in a large number of applications, such as powder coating and processing of cohesive powders. Established measurement methods such as Atomic Force Microscopy (AFM) and the centrifugal method are costly and time consuming. For engineering applications there is a need to develop a quick test method. The drop test method has been designed and developed for this purpose. In this test method particles that are adhered to a substrate are mounted on and are subjected to a tensile force by impacting the stub against a stopper ring by dropping it from a set height. From the balance of the detachment force and adhesive force for a critical particles size, above which particles are detached and below which they remain on the substrate, the interfacial specific energy is calculated. A model of adhesion is required to estimate the adhesive force between the particles and the surface, and in this work we use the JKR theory. The detachment force is estimated by Newton's second law of motion, using an estimated particle mass, based on its size and density and calculated particle acceleration. A number of materials such as silanised glass beads, Avicel, α-lactose monohydrate and starch have been tested and the adhesive force and energy between the particle and the substrate surface have been quantified. Consistent values of the interface energy with a narrow error band are obtained, independent of the impact velocity. As the latter is varied, different particle sizes detach; nevertheless similar values of the interface energy are obtained, an indication that the technique is robust, as it is in fact based on microscopic observations of many particles. The trends of the results obtained with the drop test method are similar to those shown in studies by other researchers using established methods like the AFM and the centrifuge method

Structural Characterization of Mesoporous Silica Nanofibers Synthesized Within Porous Alumina Membranes

Mesoporous silica nanofibers were synthesized within the pores of the anodic aluminum oxide template using a simple sol–gel method. Transmission electron microscopy investigation indicated that the concentration of the structure-directing agent (EO20PO70EO20) had a significant impact on the mesostructure of mesoporous silica nanofibers. Samples with alignment of nanochannels along the axis of mesoporous silica nanofibers could be formed under the P123 concentration of 0.15 mg/mL. When the P123 concentration increased to 0.3 mg/mL, samples with a circular lamellar mesostructure could be obtained. The mechanism for the effect of the P123 concentration on the mesostructure of mesoporous silica nanofibres was proposed and discussed

Self-Assembly in Monoelaidin Aqueous Dispersions: Direct Vesicles to Cubosomes Transition

Background: In the present study, synchrotron small-angle X-ray scattering (SAXS) and Cryo-TEM were used to characterize the temperature-induced structural transitions of monoelaidin (ME) aqueous dispersion in the presence of the polymeric stabilizer F127. We prove that the direct transition from vesicles to cubosomes by heating this dispersion is possible. The obtained results were compared with the fully hydrated bulk ME phase. Methodology/principal findings: Our results indicate the formation of ME dispersion, which is less stable than that based on the congener monoolein (MO). In addition, the temperature-dependence behavior significantly differs from the fully hydrated bulk phase. SAXS findings indicate a direct L(alpha)-V(2) internal transition in the dispersion. While the transition temperature is conserved in the dispersion, the formed cubosomes with internal Im3m symmetry clearly contain more water and this ordered interior is retained over a wider temperature range as compared to its fully hydrated bulk system. At 25 degrees C, Cryo-TEM observations reveal the formation of most likely closely packed onion-like vesicles. Above the lamellar to non-lamellar phase transition at 65 degrees C, flattened cubosomes with an internal nanostructure are observed. However, they have only arbitrary shapes and thus, their morphology is significantly different from that of the well-shaped analogous MO cubosome and hexosome particles. Conclusions/significance: Our study reveals a direct liposomes-cubosomes transition in ME dispersion. The obtained results suggest that the polymeric stabilizer F127 especially plays a significant role in the membrane fusion processes. F127 incorporates in considerable amount into the internal nanostructure and leads to the formation of a highly swollen Im3m phase

Paramagnetic limiting of the upper critical field of the layered organic superconductor κ−(BEDT−TTF)2Cu(SCN)2\kappa -(BEDT-TTF)_2Cu(SCN)_2

We report detailed measurements of the interlayer magnetoresistance of the layered organic superconductor $\kappa -(BEDT-TTF)_2Cu(SCN)_2$ for temperatures down to 0.5 K and fields up to 30 tesla. The upper critical field is determined from the resistive transition for a wide range of temperatures and field directions. For magnetic fields parallel to the layers, the upper critical field increases approximately linearly with decreasing temperature. The upper critical field at low temperatures is compared to the Pauli paramagnetic limit, at which singlet superconductivity should be destroyed by the Zeeman splitting of the electron spins. The measured value is comparable to a value for the paramagnetic limit calculated from thermodynamic quantities but exceeds the limit calculated from BCS theory. The angular dependence of the upper critical field shows a cusp-like feature for fields close to the layers, consistent with decoupled layers.Comment: 16 pages 5 figure