Role of outer membrane exopolymers of Acidithiobacillus ferrooxidans in adsorption of cells onto pyrite and chalcopyrite

Bacterial surface polymers play a major role in the adhesion of bacterial cells to solid surfaces. Lipopolysaccharides (LPS) are essential constituents of the cell walls of almost all Gram-negative bacteria. This paper reports the results of the investigations on the role of outer membrane exopolymers (LPS) of the chemolithotroph, Acidithiobacillus ferrooxidans, in adsorption of the cells onto pyrite and chalcopyrite. Optimization of EDTA treatment for removal of LPS from cell surface and the surface characterization of EDTA-treated cells are outlined. There was no change in cell morphology or loss in cell motility upon treatment with upto 0.04 mM EDTA for 1 h. Partial removal of LPS by EDTA treatment resulted in reduced adsorption of the cells on both pyrite and chalcopyrite. The protein profile of the EDTA-extractable fraction showed presence of certain outer membrane proteins indicating that EDTA treatment results in temporary gaps in the outer membrane. Also, specificity towards pyrite compared to chalcopyrite that was exhibited by untreated cells was lost when their exopolymer layers were stripped off, which could be attributed to the role of outer membrane proteins in the mineral-specificity exhibited by the bacteria. (C) 2013 Elsevier B.V. All rights reserved.

Direct visualization of the dynamics of membrane-anchor proteins in living cells

Dynamic properties of proteins have crucial roles in understanding protein function and molecular mechanism within cells. In this paper, we combined total internal reflection fluorescence microscopy with oblique illumination fluorescence microscopy to observe directly the movement and localization of membrane-anchored green fluorescence proteins in living cells. Total internal reflect illumination allowed the observation of proteins in the cell membrane of living cells since the penetrate depth could be adjusted to about 80 nm, and oblique illumination allowed the observation of proteins both in the cytoplasm and apical membrane, which made this combination a promising tool to investigate the dynamics of proteins through the whole cell. Not only individual protein molecule tracks have been analyzed quantitatively but also cumulative probability distribution function analysis of ensemble trajectories has been done to reveal the mobility of proteins. Finally, single particle tracking has acted as a compensation for single molecule tracking. All the results exhibited green fluorescence protein dynamics within cytoplasm, on the membrane and from cytoplasm to plasma membrane.

Part I. Influence of membrane morphology on ion-exchange and charge propagation in composite polyelectrolyte electrode coatings. Part II. Dynamic surface tension measurements of polarization relaxation at mercury pool electrodes by the method of Wilhelmy.

Part I. Novel composite polyelectrolyte materials were developed that exhibit desirable charge propagation and ion-retention properties. The morphology of electrode coatings cast from these materials was shown to be more important for its electrochemical behavior than its chemical composition.

Part II. The Wilhelmy plate technique for measuring dynamic surface tension was extended to electrified liquid-liquid interphases. The dynamical response of the aqueous NaF-mercury electrified interphase was examined by concomitant measurement of surface tension, current, and applied electrostatic potential. Observations of the surface tension response to linear sweep voltammetry and to step function perturbations in the applied electrostatic potential (e.g., chronotensiometry) provided strong evidence that relaxation processes proceed for time-periods that are at least an order of magnitude longer than the time periods necessary to establish diffusion equilibrium. The dynamical response of the surface tension is analyzed within the context of non-equilibrium thermodynamics and a kinetic model that requires three simultaneous first order processes.

On a method of determination of chlorophyll in cells of algae collected on a membrane filter. [Translation from: Trudy Instituta Biologii Vnutrennykh Vodnany 11(14), 198-202, 1966.]

The original method, proposed by Yentsch (1957), of determination of chlorophyll directly in the cells, attracts attention by its simplicity. In order to measure the content of chlorophyll by this method, a determined volume of suspension of algae is filtered through a membrane filter. The latter is dried a little, clarified by immersion oil, clamped between two glasses, and spectrophotometrized. Extinction is read off at , wavelengths equal to 670 millimicrons (around the maximum absorption of chlorophyll a in the cell) and 750 millimicrons (correction for non- specific absorption and dispersion of light by particles of the preparation). The method of Yentsch was employed by the authors for determination of chlorophyll-a in samples of phytoplankton. They conclude that in spite of the simplicity and convenience of determination the method must be applied sufficiently carefully. It is more suitable for analysis of cultures of algae, where, non-specific absorption of light is insignificant.

eta photoproduction on the proton in a chiral constituent quark approach via a one-gluon-exchange model

A formalism based on a chiral quark model (chi QM) approach complemented with a one-gluon-exchange model, to take into account the breakdown of the SU(6)circle times O(3) symmetry, is presented. The configuration mixing of wave functions for nucleon and resonances are derived. With few adjustable parameters, differential cross-section and polarized-beam asymmetry for the gamma p -> eta p process are calculated and successfully compared with the data in the center-of-mass energy range from threshold to 2 GeV. The known resonances S-11(1535), S-11(1650), P-13(1720), D-13(1520), and F-15(1680), as well as two new S-11 and D-15 resonances, are found to be dominant in the reaction mechanism. Moreover, connections among the scattering amplitudes of the chi QM approach and the helicity amplitudes, as well as decay widths of resonances, are established. Possible contributions from the so-called missing resonances are investigated and found to be negligible.

Fabrication of a miniature twin-fuel-cell on silicon wafer

The fabrication and performance evaluation of a miniature twin-fuel-cell on silicon wafers are presented in this paper. The miniature twin-fuel-cell was fabricated in series using two membrane-electrode-assemblies sandwiched between two silicon substrates in which electric current, reactant, and product flow. The novel structure of the miniature twin-fuel-cell is that the electricity interconnect from the cathode of one cell to the anode of another cell is made on the same plane. The interconnect was fabricated by sputtering a layer of copper over a layer of gold on the top of the silicon wafer. Silicon dioxide was deposited on the silicon wafer adjacent to the copper layer to prevent short-circuiting between the twin cells. The feed holes and channels in the silicon wafers were prepared by anisotropic silicon etching from the back and front of the wafer with silicon dioxide acting as intrinsic etch-stop layer. Operating on dry H-2/O-2 at 25 degreesC and atmospheric pressure, the measured peak power density was 190.4 mW/cm(2) at 270 mA/cm(2) for the miniature twin-fuel-cell using a Nafion 112 membrane. Based on the polarization curves of the twin-fuel-cell and the two single cells, the interconnect resistance between the twin cells was calculated to be in the range from 0.0113 Omega (at 10 mA/cm(2)) to 0.0150 Omega (at 300 mA/cm(2)), which is relatively low. (C) 2003 Elsevier Science Ltd. All rights reserved.

Synthesis and properties of sulfonated poly[bis(benzimidazobenzisoquinolinones)] as hydrolytically and thermooxidatively stable proton conducting ionomers

Novel sulfonated poly [bis(benzimidazobenzisoquinolinones)] as hydrolytically and thermooxidatively stable electrolyte for high -temperature fuel cell applications are reported. A series of sulfonated polymers (SPBIBI-x, x refers to molar percentage of sulfonated dianhydride monomer) were synthesized from 6,6'-disulfonic-4,4'-binaphthyl-1,1',8,8'-tetracarboxylic dianhydride (SBTDA), 4,4-binaphthyl-1,1,8,8-tetracarboxylic dianhydride (BTDA), and 3,3'-diaminobenzidine. The chemical structures of those polymers as well as model compounds synthesized from SBTDA and o-phenylenediamine were confirmed by nuclear magnetic resonance (NMR) and Fourier transform infrared (FTIR).

Proton transfer across conducting polypyrrole membrane separated by two electrolyte solutions

Electrochemical behavior of the transfer of H+ across polypyrrole membrane (PPM) was studied. The transfer process was quasi-reversible and mainly diffusion-controlled. PPM electropolymerized in water solution has better reversibility than that in CH3CN solution for the transfer of H+. The transfer process of H+ across the two kinds of PPM indicated that the PPM electrochemically polymerized was of asymmetry.

Improvement of direct methanol fuel cell performance by modifying catalyst coated membrane structure

A five-layer catalyst coated membrane (CCM) based upon Nation 115 membrane for direct methanol fuel cell (DMFC) was designed and fabricated by introducing a modified Nafion layer between the membrane and the catalyst layer. The properties of the CCM were determined by SEM, cyclic voltammetry, impedance spectroscopy, ruinous test and I-V curves. The characterizations show that the modified Nation layers provide increased interface contact area and enhanced interaction between the membrane and the catalyst layer. As a result, higher Pt utilization, lower contact resistance and superior durability of membrane electrode assembly was achieved. A 75% Pt utilization efficiency was obtained by using the novel CCM structure, whereas the conventional structure gave 60% efficiency. All these features greatly contribute to the increase in DMFC performance. The DMFC with new CCM structure presented a maximum power density of 260 MW cm(-2), but the DMFC with conventional structure gave only 200 mW cm(-2) under the same operation condition. (c) 2005 Elsevier B.V. All rights reserved.

Pendrin: the thyrocyte apical membrane iodide transporter?

In the thyroid, the transport of iodide from the extracellular space to the follicular lumen requires two steps: the transport in the cell at the basal side and in the lumen at the apical side. The first step is mediated by the Na(+)/I(-) symporter (NIS). In most reviews and textbooks, the second step is presented as mediated by pendrin. In this review, we analyze this assumption. There are several arguments supporting the concept that indeed pendrin plays an important role in thyroid physiology. However, biochemical, clinical and histological data on the thyroid of a patient with Pendred syndrome do not suggest an essential role in iodide transport, which is corroborated by the lack of a thyroid phenotype in pendrin knockout mice. Experiments in vivo and in vitro on polarized and unpolarized cells show that iodide is transported transport of iodide at the apex of the thyroid cell. Moreover, ectopic expression of pendrin in transfected non-thyroid cells is capable of mediating iodide efflux. It is concluded that pendrin may participate in the iodide efflux into thyroid lumen but not as the unique transporter. Moreover, another role of pendrin in mediating Cl(-)/HCO(3)(-) exchange and controlling luminal pH is suggested.

Membrane binding of plasmid DNA and endocytic pathways are involved in electrotransfection of mammalian cells.

Electric field mediated gene delivery or electrotransfection is a widely used method in various studies ranging from basic cell biology research to clinical gene therapy. Yet, mechanisms of electrotransfection are still controversial. To this end, we investigated the dependence of electrotransfection efficiency (eTE) on binding of plasmid DNA (pDNA) to plasma membrane and how treatment of cells with three endocytic inhibitors (chlorpromazine, genistein, dynasore) or silencing of dynamin expression with specific, small interfering RNA (siRNA) would affect the eTE. Our data demonstrated that the presence of divalent cations (Ca(2+) and Mg(2+)) in electrotransfection buffer enhanced pDNA adsorption to cell membrane and consequently, this enhanced adsorption led to an increase in eTE, up to a certain threshold concentration for each cation. Trypsin treatment of cells at 10 min post electrotransfection stripped off membrane-bound pDNA and resulted in a significant reduction in eTE, indicating that the time period for complete cellular uptake of pDNA (between 10 and 40 min) far exceeded the lifetime of electric field-induced transient pores (∼10 msec) in the cell membrane. Furthermore, treatment of cells with the siRNA and all three pharmacological inhibitors yielded substantial and statistically significant reductions in the eTE. These findings suggest that electrotransfection depends on two mechanisms: (i) binding of pDNA to cell membrane and (ii) endocytosis of membrane-bound pDNA.