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We report the novel fabrication of a highly sensitive, selective, fast responding, and affordable amperometric glucose biosensor using exfoliated graphite nanoplatelets (xGnPs) decorated with Pt and Pd nanoparticles. Nafion was used to solubilize metal-decorated graphite nanoplatelets, and a simple cast method with high content organic solvent (85 wt %) was used to prepare the biosensors. The addition of precious metal nanoparticles such as platinum (Pt) and palladium (Pd) to xGnP increased the electroactive area of the electrode and substantially decreased the overpotential in the detection of hydrogen peroxide. The Pt-xGnP glucose biosensor had a sensitivity of 61.5+/-0.6 microA/(mM x cm(2)) and gave a linear response up to 20 mM. The response time and detection limit (S/N=3) were determined to be 2 s and 1 microM, respectively. Therefore, this novel glucose biosensor based on the Pt nanoparticle coated xGnP is among the best reported to date in both sensing performance and production cost. In addition, the effects of metal nanoparticle loading and the particle size on the biosensor performance were systematically investigated.
ACS Nano 2008 Sep 23
PMID:Nanometal-decorated exfoliated graphite nanoplatelet based glucose biosensors with high sensitivity and fast response. 1920 21

A series of nanoporous membranes prepared from polyethylene-block-polystyrene were applied for size-selective diffusion of glucose and albumin molecules. Millimeter-sized test cells for characterization of such molecular diffusions were designed assuming an implantable glucose sensor. The prepared nanoporous membrane exhibits excellent flexibility and toughness compared to conventional nanoporous membranes of brittle alumina. Pore size of the membranes could be controlled from 5 to 30 nm by varying preparation conditions. All of these nanoporous membranes prepared in this study let glucose pass through, indicating a continuous pore connection through the entire thickness of the membrane in a few tens of micrometers. In contrast, membranes prepared under optimum conditions could perfectly block albumin permeation. This means that these vital molecules having different sizes can be selectively diffused through the nanoporous membranes. Such a successful combination of size selectivity of molecular diffusion in nanoscale and superior mechanical properties in macroscale is also beneficial for other devices requesting down-sized manufacture.
ACS Nano 2009 Apr 28
PMID:Size-selective diffusion in nanoporous but flexible membranes for glucose sensors. 1932 85

Many receptors undergo ligand-induced conformational changes to initiate signal transduction. Periplasmic binding proteins (PBPs) are bacterial receptors that exhibit dramatic conformational changes upon ligand binding. These proteins mediate a wide variety of fundamental processes including transport, chemotaxis, and quorum sensing. Despite the importance of these receptors, no PBP antagonists have been identified and characterized. In this study, we identify 3-O-methyl-d-glucose as an antagonist of glucose/galactose-binding protein and demonstrate that it inhibits glucose chemotaxis in E. coli. Using small-angle X-ray scattering and X-ray crystallography, we show that this antagonist acts as a wedge. It prevents the large-scale domain closure that gives rise to the active signaling state. Guided by these results and the structures of open and closed glucose/galactose-binding protein, we designed and synthesized an antagonist composed of two linked glucose residues. These findings provide a blueprint for the design of new bacterial PBP inhibitors. Given the key role of PBPs in microbial physiology, we anticipate that PBP antagonists will have widespread uses as probes and antimicrobial agents.
ACS Chem Biol 2009 Jun 19
PMID:Structure-based design of a periplasmic binding protein antagonist that prevents domain closure. 1934 66

We assessed the mechanisms by which specialized hypothalamic ventromedial nucleus (VMN) neurons utilize both glucose and long-chain fatty acids as signaling molecules to alter their activity as a potential means of regulating energy homeostasis. Fura-2 calcium (Ca(2+)) and membrane potential dye imaging, together with pharmacological agents, were used to assess the mechanisms by which oleic acid (OA) alters the activity of dissociated VMN neurons from 3- to 4-wk-old rats. OA excited up to 43% and inhibited up to 29% of all VMN neurons independently of glucose concentrations. In those neurons excited by both 2.5 mM glucose and OA, OA had a concentration-dependent effective excitatory concentration (EC(50)) of 13.1 nM. Neurons inhibited by both 2.5 mM glucose and OA had an effective inhibitory concentration (IC(50)) of 93 nM. At 0.5 mM glucose, OA had markedly different effects on these same neurons. Inhibition of carnitine palmitoyltransferase, reactive oxygen species formation, long-chain acetyl-CoA synthetase and ATP-sensitive K(+) channel activity or activation of uncoupling protein 2 (UCP2) accounted for only approximately 20% of OA's excitatory effects and approximately 40% of its inhibitory effects. Inhibition of CD36, a fatty acid transporter that can alter cell function independently of intracellular fatty acid metabolism, reduced the effects of OA by up to 45%. Thus OA affects VMN neuronal activity through multiple pathways. In glucosensing neurons, its effects are glucose dependent. This glucose-OA interaction provides a potential mechanism whereby such "metabolic sensing" neurons can respond to differences in the metabolic states associated with fasting and feeding.
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PMID:Characteristics and mechanisms of hypothalamic neuronal fatty acid sensing. 1953 76

A boron-doped diamond nanorod forest (BDDNF) electrode has been fabricated by hot filament chemical vapor deposition (HFCVD) method. This BDDNF electrode exhibits very attractive electrochemical performance compared to conventional planar boron-doped diamond (BDD) electrodes, notably improved sensitivity and selectivity for biomolecule detection. The BDDNF electrode, with the possibility of fabricating a sensitive biosensor for glucose without any catalyst or mediators, shows good activity toward direct detection of glucose by simply putting the bare BDDNF electrode into the glucose solution. Furthermore, the marked selectivity of the BDDNF electrode is very favorable for the determination of glucose in the presence of ascorbic acid (AA) and uric acid (UA). The robust sensitive and selective responses of this nanostructure indicate the promise of this kind of diamond electrode for real applications.
ACS Nano 2009 Aug 25
PMID:Fabrication of boron-doped diamond nanorod forest electrodes and their application in nonenzymatic amperometric glucose biosensing. 1962 36

Acetyl-CoA produced in mitochondria from carbohydrate or amino acid catabolism needs to reach the cytosol to initiate de novo synthesis of fatty acids. All eukaryotes analyzed so far use a citrate/malate shuttle to transfer acetyl group equivalents from the mitochondrial matrix to the cytosol. Here we investigate how this acetyl group transfer occurs in the procyclic life cycle stage of Trypanosoma brucei, a protozoan parasite responsible of human sleeping sickness and economically important livestock diseases. Deletion of the potential citrate lyase gene, a critical cytosolic enzyme of the citrate/malate shuttle, has no effect on de novo biosynthesis of fatty acids from (14)C-labeled glucose, indicating that another route is used for acetyl group transfer. Because acetate is produced from acetyl-CoA in the mitochondrion of this parasite, we considered genes encoding cytosolic enzymes producing acetyl-CoA from acetate. We identified an acetyl-CoA synthetase gene encoding a cytosolic enzyme (AceCS), which is essential for cell viability. Repression of AceCS by inducible RNAi results in a 20-fold reduction of (14)C-incorporation from radiolabeled glucose or acetate into de novo synthesized fatty acids. Thus, we demonstrate that the essential cytosolic enzyme AceCS of T. brucei is responsible for activation of acetate into acetyl-CoA to feed de novo biosynthesis of lipids. To date, Trypanosoma is the only known eukaryotic organism that uses acetate instead of citrate to transfer acetyl groups over the mitochondrial membrane for cytosolic lipid synthesis.
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PMID:Acetate produced in the mitochondrion is the essential precursor for lipid biosynthesis in procyclic trypanosomes. 1962 28

We demonstrate the use of hydrogel swelling as a mechanism to reversibly induce solvatochromic shifting in single-walled carbon nanotube (SWNT) near-infrared emission within a biocompatible hydrogel. The optical sensor reports the degree of the swelled state and glucose concentration when apo-glucose oxidase is used to cross-link the hydrogel. Photoluminescence emission maxima from dispersed nanotubes in a poly(vinyl alcohol) hydrogel shift as cross-linking is increased, with a maximum of -48 meV for the (6,5) nanotube. The Raman tangential mode also red shifts up to 17 cm(-1), indicative of nanotube lattice strain equivalent to an effective hydrostatic pressure of 3 GPa. While the electronic band gaps of SWNTs are known to either increase or decrease with uniaxial strain or lattice deformation depending on chiral vector, we show that the mechanism of detection is counterintuitively non-strain-dependent. Instead, the data are well-described by a model that accounts for changes in dielectric screening of the 1-D exciton, as the osmotic pressure forces conformational distortions in the PVA by rotating more polar groups to the nanotube surface. The model describes observed changes with hydration state and cross-linking density variation from 0 to 14%. Cross-linking with apo-glucose oxidase renders the hydrogel glucose responsive, and we demonstrate rapid and reversible detection of glucose from these systems after repeated cycling of 10 mM glucose. We also demonstrate detection and imaging in the near-infrared of implanted hydrogel sensors in a mouse tissue model, showing excellent signal-to-noise of 8.6 and contrast with integration times of 60 s.
ACS Nano 2009 Dec 22
PMID:Modulation of single-walled carbon nanotube photoluminescence by hydrogel swelling. 1992 95

Layered LiNi(1/3)Co(1/3)Mn(1/3)O2, which is isostructural to LiCoO2, is considered as a potential cathode material. A layer of carbon coated on the particles improves the electrode performance, which is attributed to an increase of the grain connectivity and also to protection of metal oxide from chemical reaction. The present work involves in situ synthesis of carbon-coated submicrometer-sized particles of LiNi(1/3)Co(1/3)Mn(1/3)O2 in an inverse microemulsion medium in the presence of glucose. The precursor obtained from the reaction is heated in air at 900 degrees C for 6 h to get crystalline LiNi(1/3)Co(1/3)Mn(1/3)O2. The carbon coating is found to impart porosity as well as higher surface area in relation to bare samples of the compound. The electrochemical characterization studies provide that carbon-coated LiNi(1/3)Co(1/3)Mn(1/3)O2 samples exhibit improved rate capability and cycling performance. The carbon coatings are shown to suppress the capacity fade, which is normally observed for the bare compound. Impedance spectroscopy data provide additional evidence for the beneficial effect of a carbon coating on LiNi(1/3)Co(1/3)Mn(1/3)O2 particles.
ACS Appl Mater Interfaces 2009 Jun
PMID:Synthesis and characterization of carbon-coated LiNi(1/3)Co(1/3)Mn(1/3)O2 in a single step by an inverse microemulsion route. 2035 19

Ag(2)O nanowalls consisting of densely packed nanoplates based on a Cu substrate were synthesized through a facile one-pot hydrothermal method. A new enzymeless glucose sensor of Cu-Ag(2)O nanowalls was fabricated. The Cu-Ag(2)O nanowalls showed higher catalysis on glucose oxidation than traditional Ag(2)O nanoflowers and Cu-Ag(2)O nanospindles. At an applied potential of 0.4 V, the sensor produced an ultrahigh sensitivity to glucose (GO) of 298.2 microA mM(-1). Linear response was obtained over a concentration range from 0.2 mM to 3.2 mM with a detection limit of 0.01 mM (S/N = 3). Satisfyingly, the Cu-Ag(2)O nanowalls modified electrode was not only successfully employed to eliminate the interferences from uric acid (UA) acid ascorbic (AA) and also fructose (FO) during the catalytic oxidation of glucose. The Cu-Ag(2)O nanowalls modified electrode allows highly sensitive, excellently selective, stable, and fast amperometric sensing of glucose and thus is promising for the future development of nonenzymatic glucose sensors.
ACS Appl Mater Interfaces 2009 Dec
PMID:Silver oxide nanowalls grown on Cu substrate as an enzymeless glucose sensor. 2035 63

This paper describes a biomaterial microfabrication approach for interfacing functional biomolecules (enzymes) with electrode arrays. Poly (ethylene glycol) (PEG) hydrogel photopatterning was employed to integrate gold electrode arrays with the enzymes glucose oxidase (GOX) and lactate oxidase (LOX). In this process, PEG diacrylate (DA)-based prepolymer containing enzyme molecules as well as redox species (vinylferrocene) was spin-coated, registered, and UV cross-linked on top of an array of gold electrodes. As a result, enzyme-carrying circular hydrogel structures (600 microm diameter) were fabricated on top of 300 microm diameter gold electrodes. Importantly, when used with multiple masks, hydrogel photolithography allowed us to immobilize GOX and LOX molecules on adjacent electrodes within the same electrode array. Cyclic voltammetry and amperometry were used to characterize biosensor electrode arrays. The response of the biosensor array was linear for up to 20 mM glucose with sensitivity of 0.9 microA cm(-2) mM(-1) and 10 mM lactate with sensitivity of 1.1 microA cm(-2) mM(-1). Importantly, simultaneous detection of glucose and lactate from the same electrode array was demonstrated. A novel strategy for integrating biological and electrical components of a biosensor described in this paper provides the flexibility to spatially resolve and register different biorecognition elements with individual members of a miniature electrode array. Of particular interest to us are future applications of these miniature electrodes for real-time monitoring of metabolite fluxes in the vicinity of living cells.
ACS Appl Mater Interfaces 2010 Mar
PMID:Immobilizing enzymes onto electrode arrays by hydrogel photolithography to fabricate multi-analyte electrochemical biosensors. 2035 76


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