Remediation of textile effluents for water reuse: Decolorization and desalination using Escherichia fergusonii followed by detoxification with activated charcoal.

Textile effluents contain high levels of pollutants of different categories like dyes, metal salts, acids, bases and microorganisms. Remediation of textile effluents is often challenging because of its composition, which also varies between dyeing units. In this study, we demonstrate the novel use of a waste-water bacterium, Escherichia fergusonii, in the effective remediation of textile effluents. The bacteria application efficiently caused a reduction of color (98.4%), total dissolved solids (75%), sulphates (87%), bicarbonates (83%), chlorides (64%), calcium (84%), and chemical oxygen demand (81%) of the textile effluents. The bacteria-treated effluents were further disinfected and detoxified by treating with rice husk activated charcoal. After the charcoal treatment, the chemical oxygen demand decreased further by 11.5% and biochemical oxygen demand decreased by 85%. The effluents remediated using the two-step process were subjected to toxicity assays using zebrafish (Danio rerio) model. The textile effluents treated using Escherichia fergusonii, followed by activated charcoal were found to be non-toxic and suitable for reuse for domestic applications. Thus, we present here, a simple, less energy-intensive, economic, two-step process as a complete solution for textile effluent treatment. The results of this investigation can be used to simplify the remediation process of textile effluents in common treatment plants as well as the individual dyeing units.

Electrochemical evidence for asialoglycoprotein receptor--mediated hepatocyte adhesion and proliferation in three dimensional tissue engineering scaffolds.

Asialoglycoprotein receptor (ASGPR) is one of the recognition motifs on the surface of hepatocytes, which promote their adhesion to extracellular matrix in liver tissue and appropriate artificial surfaces. ASGPR-mediated adhesion is expected to minimize trans-differentiation of hepatocytes in vitro that is generally observed in integrin-mediated adhesion. The aim of the present study is to verify the role of ASGPR in hepatocyte adhesion and proliferation in scaffolds for hepatic tissue engineering. Scanning Electrochemical Microscopy (SECM) is emerging as a suitable non-invasive analytical tool due to its high sensitivity and capability to correlate the morphology and activity of live cells. HepG2 cells and rat primary hepatocytes cultured in Polyvinyl alcohol (PVA)/Gelatin hydrogel scaffolds with and without galactose (a ligand for ASGPR) modification are studied using SECM. Systematic investigation of live cells cultured for different durations in scaffolds of different compositions (9:1 and 8:2 PVA:Gelatin with and without galactose) reveals significant improvement in cell-cell communication and proliferation on galactose incorporated scaffolds, thereby demonstrating the positive influence of ASGPR-mediated adhesion. In this work, we have also developed a methodology to quantify the respiratory activity and intracellular redox activity of live cells cultured in porous tissue engineering scaffolds. Using this methodology, SECM results are compared with routine cell culture assays viz., MTS ((1-Oxyl-2,2,5,5,-tetramethyl-Δ3-pyrroline-3-methyl) Methanethiosulfonate) and Albumin assays to demonstrate the better sensitivity of SECM. In addition, the present study demonstrates SECM as a reliable and sensitive tool to monitor the activity of live cells cultured in scaffolds for tissue engineering, which could be used on a routine basis.

Real-time mapping of salt glands on the leaf surface of Cynodon dactylon L. using scanning electrochemical microscopy.

Salt glands are specialized organelles present in the leaf tissues of halophytes, which impart salt-tolerance capability to the plant species. These glands are usually identified only by their morphology using conventional staining procedures coupled with optical microscopy. In this work, we have employed scanning electrochemical microscopy to identify the salt glands not only by their morphology but also by their salt excretion behavior. Bermuda grass (Cynodon dactylon L.) species was chosen for the study as they are known to be salt-tolerant and contain salt glands on leaf surfaces. Scanning electrochemical microscopy performed in sodium chloride medium in the presence and absence of potassium ferrocyanide as redox mediator, reveals the identity of salt glands. More insight into the ion expulsion behavior of these glands was obtained by mapping lateral and vertical variations in ion concentrations using surface impedance measurements which indicated five times higher resistance over the salt glands compared to the surrounding tissues and bulk solution. The protocol could be used to understand the developmental processes in plants grown in different soil/water conditions in order to improve salt tolerance of food crops by genetic engineering and hence improve their agricultural productivity.

Bio-inspired morphological evolution of zinc oxide nanostructures on a tunable enzyme platform.

Diamine oxidase is a copper-containing enzyme with interesting structural dynamics sensitive to environmental conditions. The present work explores the applicability of the system as a tunable platform for the shape and size selective synthesis of zinc oxide nanoparticles under ambient conditions. Significant changes in the nanoscale morphology of ZnO have been observed, using scanning electron microscopy, with respect to changes in pH and gas atmosphere of the medium. More specifically, hexagonal plates of nanoscale ZnO were formed at pH below the isoelectric point of the enzyme and spherical particles at alkaline pH. Interestingly the average particle size of ZnO nanostructures increases with increasing oxygen content at acidic pH while the opposite trend is noticed at alkaline pH. The observations are explained on the basis of changes in the enzyme's surface charge, conformation and redox potential using a combination of techniques like zeta potential measurements, FTIR spectroscopy, fluorescence spectroscopy, open circuit potential studies and cyclic voltammetry. Thus the present work demonstrates the applicability of an enzyme as a dynamic bio-template for the synthesis of a multitude of ZnO nanostructures which are expected to add newer insight into bottom-up fabrication of oxide nanostructures.

Nanotube-grafted polyacrylamide hydrogels for electrophoretic protein separation.

Multiwalled carbon nanotube-modified polyacrylamide gels have been employed for the electrophoretic separation of proteins. Two approaches are compared in this investigation, one using nanotubes only as fillers inside the gel matrix and the other using nanotubes as catalyst for polymerization of acrylamide. In both the cases, polymerization of acryl-amide/bisacrylamide has been carried out in situ in the presence of nanotubes dispersed in the gel buffer containing monomer and cross-linker. In the former case, initiator and catalyst have been added after ultrasonication of nanotubes in the gel buffer mixture where the nanotubes play the role of filler. On the other hand, the second approach precludes use of catalyst and involves addition of initiator alone during ultrasonication of nanotubes in the gel buffer containing monomer and cross-linker, which leads to the formation of nanotube-grafted gel after 25 min. When nanotubes are used as a catalyst instead of N,N,N',N'-tetramethylethylenediamine, pore size distribution of the gel matrix and linearity of molecular weight calibration plots are found to be improved. In addition, other issues associated with the use of an external catalyst like handling the moisture-sensitive and corrosive reagent and associated irreproducibility are addressed in this approach.

Carbon nanotube-modified sodium dodecyl sulfate-polyacrylamide gel electrophoresis for molecular weight determination of proteins.

The effect of incorporating carbon nanotubes (CNTs) in the gel matrix on the electrophoretic mobility of proteins based on their molecular weight differences was investigated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). More specifically, a reduction in standard deviation in the molecular weight calibration plots by 55% in the case of multiwalled carbon nanotubes (MWCNTs) and by 34% in the case of single-walled carbon nanotubes (SWCNTs) compared with that of pristine polyacrylamide gels was achieved after incorporating an insignificant amount of functionalized CNTs into the gel matrix. A mechanism based on a more uniform pore size distribution in CNT modified polyacrylamide gel matrix is proposed. Furthermore, the impact of SWCNTs and MWCNTs on the mobility of proteins in different molecular weight regimes at a given acrylamide concentration offers a tunable gel matrix in terms of the selection of molecular weight ranges of proteins. The robustness and excellent reproducibility of the CNT-PAGE protocol are expected to have a significant impact on the molecular weight determination of newly isolated proteins.

Domain size manipulation of perflouorinated polymer electrolytes by sulfonic acid-functionalized MWCNTs to enhance fuel cell performance.

The application of sulfonic acid-functionalized multiwalled (s-MWNT) carbon nanotubes to manipulate the hydrophilic domain size of Nafion membranes is explored here as an option for tuning the proton conductivity of polymer electrolyte membranes for hydrogen-oxygen fuel cells. The electrochemical impedance experiments provide preliminary evidence of increased proton conductivity, while small-angle X-ray scattering measurements line out enhanced ionic cluster domain size in these composite membranes as the central reason for higher conductivity (70 A for the optimum composite membrane vs 50 A for Nafion 115) values. Scanning electrochemical microscopy indicates synergistic interaction between the sulfonic acid functional groups present in the Nafion membrane and those on the nanotube surface. More interestingly, the nanotube-tailored Nafion membranes ameliorate the performance of fuel cells as confirmed by measurements at a single-cell level, which reveal a maximum power density of 380 mW cm(-2), higher than those of Nafion 115 (250 mW cm(-2)) and recast Nafion (230 mW cm(-2)) membranes. Thus, in addition to providing an elegant means of controlling the ionic cluster size, the strategic approach of using CNT both as an anchoring backbone for -SO(3)H groups to enrich proton conductivity and as a blending agent to improve the mechanical characteristics of the Nafion phase might be helpful in alleviating many critical problems associated with the use of commercial Nafion membranes.

Imaging the stomatal physiology of somatic embryo-derived peanut leaves by scanning electrochemical microscopy.

The stomatal physiology, chlorophyll distribution and photosynthetic activity of somatic embryo (SE)- and seedling-derived peanut plants grown in vitro (test tube-grown) and extra vitrum (soil-grown) are investigated using scanning electrochemical microscopy (SECM). This SECM imaging is performed in two different feedback modes, corresponding to oxygen evolution and chlorophyll distribution. More specifically, the oxygen evolution profiles of the in vitro leaves indicate important differences in leaf anatomy between the SE- and seedling-derived leaves. On the other hand, the chlorophyll distribution images show individual stomata of size ca. 27 +/- 5 microm. Further studies on senescing (aged) leaves reveal interesting voltammograms that vary widely over the stomatal complexes and the surrounding tissues, probably due to the release of electroactive metabolites during chlorophyll breakdown when the leaves turn yellow. Thus, the present investigation could open up new opportunities for characterizing botanical systems using electroanalytical techniques. In addition, it could provide further insights into various areas of current relevance, including signal transduction, cell fate/differentiation and developmental biology.

'All-solid-state' electrochemistry of a protein-confined polymer electrolyte film.

Interfacial redox behavior of a heme protein (hemoglobin) confined in a solid polymer electrolyte membrane, Nafion (a perfluoro sulfonic acid ionomer) is investigated using a unique 'all-solid-state' electrochemical methodology. The supple phase-separated structure of the polymer electrolyte membrane, with hydrophilic pools containing solvated protons and water molecules, is found to preserve the incorporated protein in its active form even in the solid-state, using UV-visible, Fluorescence (of Tryptophan and Tyrosine residues) and DRIFT (diffuse reflectance infrared Fourier transform) spectroscopy. More specifically, solid-state cyclic voltammetry and electrochemical impedance of the protein-incorporated polymer films reveal that the Fe2+-form of the entrapped protein is found to bind molecular oxygen more strongly than the native protein. In the 'all-solid-state' methodology, as there is no need to dip the protein-modified electrode in a liquid electrolyte (like the conventional electrochemical methods), it offers an easier means to study a number of proteins in a variety of polymer matrices (even biomimetic assemblies). In addition, the results of the present investigation could find interesting application in a variety of research disciplines, in addition to its fundamental scientific interest, including protein biotechnology, pharmaceutical and biomimetic chemistry.