Structure-function relationships in Anabaena ferredoxin: correlations between X-ray crystal structures, reduction potentials, and rate constants of electron transfer to ferredoxin:NADP+ reductase for site-specific ferredoxin mutants

A combination of structural, thermodynamic, and transient kinetic data on wild-type and mutant Anabaena vegetative cell ferredoxins has been used to investigate the nature of the protein-protein interactions leading to electron transfer from reduced ferredoxin to oxidized ferredoxin:NADP+ reductase (FNR). We have determined the reduction potentials of wild-type vegetative ferredoxin, heterocyst ferredoxin, and 12 site-specific mutants at seven surface residues of vegetative ferredoxin, as well as the one- and two-electron reduction potentials of FNR, both alone and in complexes with wild-type and three mutant ferredoxins. X-ray crystallographic structure determinations have been carried out for six of the ferredoxin mutants. None of the mutants showed significant structural changes in the immediate vicinity of the [2Fe-2S] cluster, despite large decreases in electron-transfer reactivity (for E94K and S47A) and sizable increases in reduction potential (80 mV for E94K and 47 mV for S47A). Furthermore, the relatively small changes in Calpha backbone atom positions which were observed in these mutants do not correlate with the kinetic and thermodynamic properties. In sharp contrast to the S47A mutant, S47T retains electron-transfer activity, and its reduction potential is 100 mV more negative than that of the S47A mutant, implicating the importance of the hydrogen bond which exists between the side chain hydroxyl group of S47 and the side chain carboxyl oxygen of E94. Other ferredoxin mutations that alter both reduction potential and electron-transfer reactivity are E94Q, F65A, and F65I, whereas D62K, D68K, Q70K, E94D, and F65Y have reduction potentials and electron-transfer reactivity that are similar to those of wild-type ferredoxin. In electrostatic complexes with recombinant FNR, three of the kinetically impaired ferredoxin mutants, as did wild-type ferredoxin, induced large (approximately 40 mV) positive shifts in the reduction potential of the flavoprotein, thereby making electron transfer thermodynamically feasible. On the basis of these observations, we conclude that nonconservative mutations of three critical residues (S47, F65, and E94) on the surface of ferredoxin have large parallel effects on both the reduction potential and the electron-transfer reactivity of the [2Fe-2S] cluster and that the reduction potential changes are not the principal factor governing electron-transfer reactivity. Rather, the kinetic properties are most likely controlled by the specific orientations of the proteins within the transient electron-transfer complex.     

Stability and thermal conductivity of nanofluids of tin dioxide synthesized via microwave-induced combustion route

SnO₂ nanofluids were prepared by dispersing tin dioxide nanoparticles in deionized (DI) water as a base fluid. 4–5 nm tin dioxide crystals were synthesized via chloride solution combustion synthesis (CSCS) using SnCl₄ and sorbitol as a novel precursor and the fuel, respectively. Ammonium nitrate was also used as the combustion aid. The molar ratio of sorbitol plus ammonium nitrate to SnCl₄ was set at unity; whereas, the molar ratio of sorbitol-to-ammonium nitrate divided by that of stoichiometric value (Φ) was varied in the range of 0.5–1.4 in order to find the optimum values of specific surface area for the CSCS technique. Transition electron microscopy (TEM), scanning electron microscopy (SEM), powder X-ray diffraction (XRD), and Brunauer–Emmet–Teller (BET) techniques were employed for the characterization of the nanoparticles. Since SnO₂ nanoparticles form clusters within fluids, the fluids were ultrasonicated to improve the dispersion and stability of the nanoparticles. The colloidal stability of the SnO₂ nanofluids was quantitatively characterized by UV–vis spectrophotometric measurements. The results of the UV–vis experiments indicate higher dispersion together with enhanced stability for the nanofluid prepared by SnO₂ nanoparticles synthesized at Φ = 1.0. After 500 h sedimentation time, the relative concentration of the nanofluid with the highest stability is remained at around 77% of the initial concentration of the fluid.A transient hot-wire apparatus was used to measure the thermal conductivities of the nanofluids. In addition, the effects of pH and temperature on the thermal conductivity were also investigated. At 353 K, for the nanofluid prepared by SnO₂ nanoparticles synthesized at Φ = 1.0 at a weight fraction of 0.024%, thermal conductivity is enhanced up to about 8.7%, with an optimal pH = 8.     

Constant flow-driven microfluidic oscillator for different duty cycles

This paper presents microfluidic devices that autonomously convert two constant flow inputs into an alternating oscillatory flow output. We accomplish this hardware embedded self-control programming using normally closed membrane valves that have an inlet, an outlet, and a membrane-pressurization chamber connected to a third terminal. Adjustment of threshold opening pressures in these 3-terminal flow switching valves enabled adjustment of oscillation periods to between 57 and 360 s with duty cycles of 0.2-0.5. These values are in relatively good agreement with theoretical values, providing the way for rational design of an even wider range of different waveform oscillations. We also demonstrate the ability to use these oscillators to perform temporally patterned delivery of chemicals to living cells. The device only needs a syringe pump, thus removing the use of complex, expensive external actuators. These tunable waveform microfluidic oscillators are envisioned to facilitate cell-based studies that require temporal stimulation.     

Development of an interactive computer-based learning strategy to assist in teaching water quality modelling

The following paper presents a computer-based learning strategy to assist in introducing and teaching water quality modelling to undergraduate civil engineering students. As part of the learning strategy, an interactive computer-based instructional (CBI) aid was specifically developed to assist students to set up, run and analyse the output from a commercially available water quality model (WQMAP) for a hypothetical raw sewage spill. The CBI aid comprised a Hypertext Markup Language (HTML) module with helpful text and still graphics, and links to on-screen recorded motion movie clips showing on-screen actions and voice narrations during the instructors absence. An anonymous class questionnaire to assess the students’ perceptions of the developed approach found that most students felt they were able to complete the project with minimum supervision, and have acquired a basic understanding of water quality modelling as a result of the activity. Additionally, almost all of the students found the CBI aid helpful and easy to follow.     

An interfacial instability in a transient wetting layer leads to lateral phase separation in thin spin-cast polymer-blend films

Spin-coating is a very widely used technique for making uniform thin polymer films. For example, the active layers in most experimental semiconducting polymer-based devices, such as light-emitting diodes and photovoltaics, are made this way. The efficiency of such devices can be improved by using blends of polymers; these phase separate during the spin-coating process, creating the complex morphology that leads to performance improvements. We have used time-resolved small-angle light scattering and light reflectivity during the spin-coating process to study the development of structure directly. Our results provide evidence that a blend of two polymers first undergoes vertical stratification; the interface between the stratified layers then becomes unstable, leading to the final phase-separated thin film. This has given us the basis for establishing a full mechanistic understanding of the development of morphology in thin mixed polymer films, allowing a route to the rational design of processing conditions so as to achieve desirable morphologies by self-assembly.     

Graphene-silica composite thin films as transparent conductors

Transparent and electrically conductive composite silica films were fabricated on glass and hydrophilic SiOx/silicon substrates by incorporation of individual graphene oxide sheets into silica sols followed by spin-coating, chemical reduction, and thermal curing. The resulting films were characterized by SEM, AFM, TEM, low-angle X-ray reflectivity, XPS, UV-vis spectroscopy, and electrical conductivity measurements. The electrical conductivity of the films compared favorably to those of composite thin films of carbon nanotubes in silica.     

The utilization of recycled concrete aggregate to produce controlled low-strength materials without using Portland cement

This paper reports the results of an experimental study that investigated the feasibility of using fine and coarse recycled concrete aggregate (RCA) with slag or fly ash to produce Controlled Low-Strength Materials (CLSM). The main objective was to produce CLSM using only recycled and by-product materials without the need to add Portland cement. In addition to the hydraulic activity of slag and high-calcium fly ash (HCFA), their pozzolanic reaction was activated by the alkalis and calcium hydroxide present in the residual paste of the RCA. Preliminary tests showed mixtures with slag to have 7-day compressive strengths 70% higher than mixtures with fly ash.Two types of CLSM with slag were investigated in further detail: one with fine and the other with fine/coarse RCA. The results showed that the developed CLSMs are suitable for a wide range of applications particularly those requiring structural support and fast hardening.     

Plant-Growth-Promoting Rhizobacteria Emerging as an Effective Bioinoculant to Improve the Growth,Production, and Stress Tolerance of Vegetable Crops

Vegetable cultivation is a promising economic activity, and vegetable consumption is important for human health due to the high nutritional content of vegetables. Vegetables are rich in vitamins, minerals, dietary fiber, and several phytochemical compounds. However, the production of vegetables is insufficient to meet the demand of the ever-increasing population. Plant-growth-promoting rhizobacteria (PGPR) facilitate the growth and production of vegetable crops by acquiring nutrients, producing phytohormones, and protecting them from various detrimental effects. In this review, we highlight well-developed and cutting-edge findings focusing on the role of a PGPR-based bioinoculant formulation in enhancing vegetable crop production. We also discuss the role of PGPR in promoting vegetable crop growth and resisting the adverse effects arising from various abiotic (drought, salinity, heat, heavy metals) and biotic (fungi, bacteria, nematodes, and insect pests) stresses.     

Effect of processing parameters on copper powder produced by novel hybrid atomisation technique

ABSTRACT

In the present work, a novel design of centrifugal atomiser for producing pure copper powder was studied. The novel complementary hybrid system provides an external stream of gas to increase the cooling rate of the atomised particles. Effects of the operating parameters, such as disc rotating speed and gas flowrate on the morphology, particle size distribution, cooling rate and microstructure, were analysed. It was evidenced from the experimental results that the median particle size in the novel atomisation process is mainly controlled by centrifugal disintegration. The microstructure of the produced powders was equiaxed and the grain size decreased with increasing gas flowrate. The cooling rate experienced by centrifugal atomised Cu powders was studied via numerical formulation estimated to be 10⁴–10^6?K?s^?1. The results show that the cooling rate is a strong function of particle size and increasing the rotating disc speed also increased the cooling rates.     

Nucleic-Acid Structures as Intracellular Probes for Live Cells

The chemical composition of cells at the molecular level determines their growth, differentiation, structure, and function. Probing this composition is powerful because it provides invaluable insight into chemical processes inside cells and in certain cases allows disease diagnosis based on molecular profiles. However, many techniques analyze fixed cells or lysates of bulk populations, in which information about dynamics and cellular heterogeneity is lost. Recently, nucleic-acid-based probes have emerged as a promising platform for the detection of a wide variety of intracellular analytes in live cells with single-cell resolution. Recent advances in this field are described and common strategies for probe design, types of targets that can be identified, current limitations, and future directions are discussed.