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PubMed2004

Electrochemical study of the XNA on Gold microarray.

A novel electrode array was developed based on the XNA on Gold trade mark microarray platform. The platform combines self-assembling monolayers, thick film patterning and streptavidin based immobilization to provide a robust, versatile platform capable of analysing virtually any biomolecule including nucleic acids, proteins, carbohydrates and lipids. Electrochemical analysis of the self-assembling monolayer/streptavidin (SAMS) XNA on Gold coating revealed that the ferrocene redox current for the SAMS modified electrode was greater than that with a bare Gold electrode. The electrochemical reaction of K4Fe(CN)6 was inhibited by the SAMS coating, but was reactivated upon addition of ferrocene. These results indicate that ferrocene is involved as a mediator in the electron transfer of K4Fe(CN)6 to the SAMS modified electrode. Addition of DNA to the SAMS resulted in only a minor change in the electrochemical signal, indicating that XNA on Gold can be used for electrochemical based bioanalysis. After cycling a SAMS electrode 50 times, no signs of deterioration were detected showing that coating has excellent stability. In addition to the biosensing applications, the scheme provides a non-invasive method for accessing the quality of the SAMS coatings which is of industrial interest. These studies show that the XNA on Gold microarray platform can be used for electrochemical studies, thus providing an additional alternative for developing multianalyte biosensors as well as expanding the range of detection methods available for microarray analysis.

DNA↗
PubMed2005

Surface modification of silicon and gold-patterned silicon surfaces for improved biocompatibility and cell patterning selectivity.

Clean silicon and gold-patterned silicon platforms were modified with methoxy-polyethylene glycol (M-PEG silane) via a self-assembly technique, which significantly improved their plasma protein resistance capability and cell patterning selectivity. Fibrinogen and IgG were used as model plasma proteins to study the efficacy of PEG layers in resisting protein adsorption. Selective cell patterning on the gold regions of a gold-patterned silicon substrate and tissue compatibility were studied with macrophage and fibroblast cells. The research also revealed how the presence of gold electrodes on a silicon substrate would influence the cell patterning selectivity. Our experimental results showed that the PEG-modified silicon surfaces had a high resistivity to protein and cell attachment and that the PEG-modified gold-patterned silicon surfaces nearly completely eliminated the protein adsorption and cell attachment on silicon. This study provides a new approach to developing biocompatible surfaces for silicon-based BioMEMS devices, particularly for biosensors where a metal-insulator format must be enforced.

Adsorption↗
PubMed2004

Direct electron transfer between cytochrome P450scc and gold nanoparticles on screen-printed rhodium-graphite electrodes.

This paper is concerned with an investigation of electron transfer between cytochrome P450scc (CYP11A1) and gold nanoparticles immobilised on rhodium-graphite electrodes. Thin films of gold nanoparticles were deposited onto the rhodium-graphite electrodes by drop casting. Cytochrome P450scc was deposited onto both gold nanoparticle modified and bare rhodium-graphite electrodes. Cyclic voltammetry indicated enhanced activity of the enzyme at the gold nanoparticle modified surface. The role of the nanoparticles in mediating electron transfer to the cytochrome P450scc was verified using ac impedance spectroscopy. Equivalent circuit analysis of the impedance spectra was performed and the values of the individual components estimated. On addition of aliquots of cholesterol to the electrolyte bioelectrocatalytic reduction currents were obtained. The sensitivity of the nanoparticle modified biosensor to cholesterol was 0.13 microA microM-1 in a detection range between 10 and 70 microM of cholesterol. This confirms that gold nanoparticles enhance electron transfer to the P450scc when present on the rhodium-graphite electrodes.

Biosensing Techniques↗
PubMed2006

Electrochemical genosensors for biomedical applications based on gold nanoparticles.

Two gold nanoparticles-based genomagnetic sensors designs for detection of DNA hybridization are described. Both assays are based on a magnetically induced direct electrochemical detection of gold tags on magnetic graphite-epoxy composite electrodes. The first design is a two strands assay format that consists of the hybridization between a capture DNA strand which is linked with paramagnetic beads and another DNA strand related to BRCA1 breast cancer gene used as a target which is coupled with streptavidin-gold nanoparticles. The second genomagnetic sensor design is a sandwich assay format with more application possibilities. A cystic fibrosis related DNA strand is used as a target and sandwiched between two complementary DNA probes: the first one linked with paramagnetic beads and a second one modified with gold nanoparticles via biotin-streptavidin complexation reactions. The electrochemical detection of gold nanoparticles by differential pulse voltammetry was performed in both cases. The developed genomagnetic sensors provide a reliable discrimination against noncomplementary DNA as well against one and three-base mismatches. Optimization parameters affecting the hybridization and analytical performance of the developed genosensors are shown for genomagnetic assays of DNA sequences related with the breast cancer and cystic fibrosis genes.

Biosensing Techniques↗
PubMed2005

Gold nanoparticles as a colorimetric sensor for protein conformational changes.

Spherical gold nanoparticles and flat gold films are prepared in which yeast iso-1-cytochrome c (Cyt c) is covalently bound to the gold surface by a thiol group in the cystein 102 residue. Upon exposure to solutions of different pH, bound Cyt c unfolds at low pH and refolds at high pH. This conformational change causes measurable shifts in the color of the coated nanoparticle solutions detected by UV-VIS absorption spectroscopy and in the refractive index (RI) of the flat gold films detected by surface plasmon resonance (SPR) spectroscopy. Both experiments demonstrate the same trend with pH, suggesting the use of protein-covered gold nanoparticles as a simple colorimetric sensor for conformational change.

Colorimetry↗
PubMed2005

Simultaneous determination of palladium, platinum, rhodium and gold by on-line solid phase extraction and high performance liquid chromatography with 5-(2-hydroxy-5-nitrophenylazo)thiorhodanine as pre-column derivatization regents.

In this paper, 5-(2-hydroxy-5-nitrophenylazo)thiorhodanine (HNATR) was synthesized. A new method for the simultaneous determination of palladium, platinum, rhodium and gold ions as metal-HNATR chelates was developed using a rapid analysis column high performance liquid chromatography equipped with on-line solid phase extraction technique. The samples (Water, human urine, geological samples and soil) were digested by microwave acid-digestion. The palladium, platinum, rhodium and gold ions in the digested samples were pre-column derivatized with HNATR to form colored chelates. The Pd-HNATR, Pt-HNATR, Rh-HNATR and Au-HNATR chelates can be absorbed onto the front of the enrichment column when they were injected into the injector and sent to the enrichment column [Zorbax Stable Bound, 10 mm x 4.6 mm, 1.8 microm] with a buffer solution of 0.05 mol L(-1) phosphoric acid as mobile phase. After the enrichment had finished, by switching the six ports switching valve, the retained chelates were back-flushed by mobile phase and travelling towards the analytical column. These chelates separation on the analytical column [Zorbax Stable Bound, 10 mm x 4.6 mm, 1.8 microm] was satisfactory with 72% acetonitrile (containing 0.05 mol L(-1) of phosphoric acid and 0.1% of Triton X-100) as mobile phase. The palladium, platinum, rhodium and gold chelates were separated completely within 2.5 min. Compared to the routine chromatographic method, more then 80% of separation time was shortened. By on-line solid phase extraction system, a large volume of sample (10 mL) can be injected, and the sensitivity of the method was greatly improved. The detection limits (S/N=3, the sample injection volume is 10 mL) of palladium, platinum, rhodium and gold in the original samples reaches 1.4, 1.8, 2.0 and 1.2 ng L(-1), respectively. The relative standard deviations for five replicate samples were 2.4-3.6%. The standard recoveries were 88-95%. This method was applied to the determination of palladium, platinum, rhodium and gold in human urine, water and geological samples with good results.

Chromatography, High Pressure Liquid↗
PubMed2005

Preparation of gold colloid monolayer by immunological identification.

The design and initial characterization of the self-assembled gold colloid monolayer by a sandwich structure via the immunological identification are reported. The 13 nm gold colloid nanoparticles and the silicon or quartz substrates have been modified with the mouse polyclonal antibody against hepatitis B virus surface antigen (PAb) and the mouse monoclonal antibody against hepatitis B virus surface antigen (MAb), respectively. They can be linked by a special reaction with their corresponding hepatitis B virus surface antigen (Antigen) as a sandwich structure. Thus, the density of gold nanoparticles self-assembled on the substrate can be readily controlled by the amount of the antigen added. The resulting substrates have been characterized by atomic force microscopy (AFM) and surface-enhanced Raman scattering (SERS) spectroscopy when the gold nanoparticles were modified with SERS-active probe molecules of 4-mercaptobenzoic acid (MBA) after silver enhancement. These data show that the gold nanoparticles are separately fixed onto the substrate and form a uniform monolayer, which possess a set of features that make them very attractive for both basic and applied uses, including roughness, high stability, and biocompatibility.

Animals↗
PubMed2004

Preparation of multilayered gold-silica-polystyrene core-shell particles by seeded polymerization.

A preparation method for multilayered gold-silica-polystyrene core-shell composite particles is proposed. The gold-silica core-shell particles of 192-nm-sized, synthesized by coating the 18-nm-sized gold particles with silica by a seeded growth technique, were used as cores for succeeding polystyrene coating. After surface modification of gold-silica composite particles by methacryloxypropyltrimethoxysilane (MPTMS), polymerizations of styrene (0.16-0.4 M) were conducted with 8 x 10(-3) M of potassium persulfate initiator in the presence of 1 x 10(-3) M of sodium p-styrenesulfonate anionic monomer. Multilayered core-shell gold-silica-polystyrene particles that contained a single core could be obtained. The coefficient of variation of size distribution (CV) of the composite particles was less than 7%, and polystyrene shell thickness was in a range of 193 to 281 nm.

Gold↗
PubMed2006

Synthesis of the gold nanodumbbells by electrochemical method.

The gold nanoparticles with novel dumbbells-like structures have been successfully fabricated by an electrochemical method using micelle template formed by two surfactants, the primary surfactant being cetyltrimethylammonium bromide (CTABr) and the co-surfactant being tetradecyltrimethylammonium bromide (TTABr). The shape of gold nanoparticles can be modified to form dumbbell structure by addition of acetone solvent during the electrolysis. The gold nanodumbbells have been determined to be single-crystalline with a face-centered cubic (FCC) structure by X-ray diffraction (XRD) analysis. The lattice constant calculated from this selected-area electron diffraction (SAED) pattern is 4.068 A. Transmission electronic microscopy (TEM) suggests that shape of gold nanodumbbell is fatter in two ends and thinner in the middle section to compare with the normal nanorod shape. These gold nanodumbbells exhibit aspect ratio (gamma) of around 3.

Acetone↗
PubMed2004

Alpha bungarotoxin-1.4 nm gold: a novel conjugate for visualising the precise subcellular distribution of alpha 7* nicotinic acetylcholine receptors.

Alpha 7 subunit-containing nicotinic acetylcholine receptors (alpha7* nAChR) are involved in a variety of functions in the mammalian brain, including modulating neurotransmitter release and synaptic plasticity. Identifying the precise cellular distribution of alpha7* nAChRs with respect to the local neurochemical environment is crucial to understanding these biological roles. Current strategies for localising alpha7* nAChRs at the subcellular level have limitations. Anti-alpha7 subunit antibodies detect both assembled and unassembled subunits whereas biotinylated alphabungarotoxin (alphaBgt) only binds to assembled alpha7* nAChRs, but interpretation of labelling is marred by co-detection of endogenous tissue biotin. To overcome these problems, we have characterised a novel 1.4 nm gold alphaBgt conjugate used to directly localise alpha7* nAChR. Gold conjugation does not significantly decrease binding affinity, and gold alphaBgt specifically labels alpha7* nAChR in both unfixed and aldehyde-fixed tissue at the light and electron microscope levels, labelling being abolished in the presence of excess competing toxin. At the ultrastructural level, gold alphaBgt is associated with neuronal membranes and located at axon-dendritic synapses in the rat hippocampus CA1 stratum radiatum. These results reveal gold alphaBgt to be a valuable new tool in elucidating the functional neuroanatomy of alpha7* nAChR in the central nervous system.

Animals↗
PubMed2006

Influence of saline and pH on collagen type I fibrillogenesis in vitro: fibril polymorphism and colloidal gold labelling.

We have produced different collagen type I fibrils by in vitro fibrillogenesis of acetic acid-soluble collagen within the pH range 2.5-9.0, in the presence and absence of 150 mM NaCl. The varying relatively stable molecular assemblies and polymorphic fibrillar end-products produced after 24 h incubation have been assessed and compared by the TEM study of specimens negatively stained with uranyl acetate. In the presence of 150 mM NaCl, the assembly of collagen at low pH (2.5) leads to the formation of initial molecular aggregates that progressively link together at slightly higher pH (5.0) to form sub-fibrils and spindle-shaped D-banded bundles of sub-fibrils. At pH 6.0 these D-banded bundles aggregate into larger spindle-shaped fibrils with lateral misalignment of the D-banding across the bundle. However, at pH 7.0 and 8.0, in the presence of 150 mM NaCl, the characteristic parallel-sided mature D-banded collagen type I fibres are formed. At pH 9.0 more loosely formed parallel-sided D-banded collagen fibrils are present, within which the spindle-shaped sub-fibrils can be defined by negative staining more convincingly than at pH 7-8. In the presence of 50 mM buffer at pH 2.5, but absence of 150 mM NaCl, collagen type I forms disorganized periodic initial molecular aggregates, which have a tendency to link together to form sub-fibrils. Flexuous collagen type I sub-fibrils predominate at pH 5.0, alongside large spindle-shaped fibrils that possess a regular transverse approximately 10 nm periodicity, with an oblique approximately 67 nm periodicity, significantly different to the D-banding periodicity. At pH 7.0 and pH 8 in the absence of saline loosely-formed flexuous and spindle-shaped fibres co-exist, with underlying sub-fibrils visible, but at pH 9.0 only disorganized flexuous fibrillar aggregates are present. Colloidal gold labelling of the characteristic D-banded collagen type I fibrils with 5 nm and 2 nm chemically reactive gold particles reveals a periodic labelling pattern, which is not apparent with 10 nm and 15 nm gold particles, due to steric hindrance. The flexuous and spindle-shaped collagen fibrils also bind 2 nm gold particles in a specific manner. In all cases, the specific chemisorption of gold onto the collagen fibrils is probably determined by the availability of repeating amino acid side chains of the collagen molecules along the fibril surface. The controlled production of varying stable collagen type I fibrillogenesis products is likely to be of value within numerous areas of biotechnology, biology and medicine, including experimental biomineralization.

Animals↗
PubMed2003

Hyper-Rayleigh scattering of protein-modified gold nanoparticles.

The nonlinear optical properties of protein-modified gold nanoparticles has been studied by the hyper-Rayleigh scattering (HRS) technique. HRS signals from the nanoparticles coated with goat-anti-human IgG have been obtained when pumped with a laser pulse with a wavelength of 1064 nm. The HRS signals of gold nanoparticles with IgG were larger than those of bare gold nanoparticles. This can be explained by a noncentrosymmetric effect. It was also found that the HRS signals from the IgG-coated gold nanoparticles could be greatly increased when the antigen was added due to gold nanoparticle aggregation. Our experiment found that the HRS method could produce a measurable signal with 10 microg/ml antigen added, while the colorimetric method using UV spectrum detection required 100 microg/ml of added antigen. The results show that the HRS measurement of immunogold nanoparticles could become a potential immunoassay in determining small levels of antigen in aqueous samples.

Animals↗
PubMed1984

A comparison of the strength of base metal and gold solder joints.

The purpose of this study was to evaluate the procedures involved in forming high- and low-fusing solder joints between two different base metal alloys and to compare the strength of these joints with the strength of those formed between specimens made from a high-gold content alloy. The conclusions of this investigation can be summarized as follows. All but 11 of the 120 specimens failed through the solder joint, which confirms that the solder joint is usually the weakest part of a fixed partial denture. The high-fusing joints formed between the base metal alloys showed higher relative bending strength values than the high-fusing joints formed between the gold alloy specimens. The low-fusing joints formed between the base metal alloys showed higher relative bending strength values than the low-fusing joints formed between the gold alloy specimens, and the low-fusing base metal joints exhibited the highest strength values in this study. The low-fusing joints formed between gold alloy specimens consistently bent before fracturing. Solder joints fail both adhesively (separation of the solder from the parent metal) and cohesively (fracture through the solder, parent metal, or a combination of both). A correctly formed joint between gold components should show cohesive fracture, whereas adhesive fractures are apparently acceptable with base metal alloys since the low-fusing base metal joints failed in this manner but still exhibited high strength values. Voids of different sizes and locations were observed in most of the solder joints, and within each group tested there was usually less strength exhibited by the joints with larger areas of porosity.

Dental Alloys↗
PubMed1997

Tensile strength of soldered gold alloy joints.

STATEMENT OF PROBLEM: Little information is available about the mechanical properties of soldered gold alloys after they have undergone various heat treatments. PURPOSE: This study investigated the influence of heat treating on the strength properties of soldered joints of two gold alloys (NC Type IV and Sofard), which can be age-hardened at intraoral temperature. MATERIAL AND METHODS: Dumbbell-shaped specimens were cast with each gold alloy and were cut at the center of the connecting bar for soldering. The two halves of the casting were then soldered with two different gold-based solders (Maingold and Degulor). Three different heat treatments were performed on the soldered gold alloy assemblies: solution heat treatment (ST) at 700 degrees C for 5 minutes (treatment A); aging at 37 degrees C for 7 days after ST (treatment B); and aging at 250 degrees C (Sofard) or 400 degrees C (NC type IV) for 15 minutes after ST (treatment C). The tensile strength, elongation and microhardness were evaluated after each heat treatment. RESULTS: The hardness values of Sofard significantly (p < 0.05) increased during aging at 37 degrees C (treatment B) and produced adequate strengths of the soldered joints, especially with the harder solder (645.7 MPa: Degulor [222 VHN] vs. 493.3 MPa: Maingold [165 VHN]). CONCLUSIONS: The results of this study indicated the possibility of strengthening soldered joints in the oral environment, thus eliminating the necessity for any additional hardening heat treatment.

Analysis of Variance↗
PubMed2002

Sensitization potential of gold sodium thiosulfate in mice and guinea pigs.

Since gold sodium thiosulfate (GST) has been included in a standard patch test series for diagnosis of allergic contact dermatitis from gold, the incidence of patients showing positive reactions to gold is increasing. However, there were little reports on induction of gold sensitization in animals. In this study, we have examined the sensitization potential of GST using mice and guinea pigs. In the guinea pig maximization test, 2 or 6 out of 10 animals showed positive skin responses, mainly edema, by challenge with 2% or 5% GST in 50% ethanol solution, respectively. In the mouse ear swelling test, positive ear swelling (20% greater increase in ear thickness) after challenge with GST was shown in 2 out of 6 mice those previously treated with GST. Topical exposure of mice to GST in 70% dimethylsulfoxide solution induced small increases in the lymph node weight and the lymph node cell (LNC) number in the murine local lymph node assay (LLNA). A greater degree of LNC responses were observed in the sensitive mouse lymph node assay (SLNA) compared with the LLNA, but the stimulation index of total lymph node response by GST was not so high. From these results, GST was identified as a contact allergen, but the sensitization potential was not so strong. In the mouse IgE test, treatment of mice with GST resulted in a statistically significant increase in the serum IgE antibody concentration that associated with immediate-type hypersensitivity reaction. It may suggest that the sensitization responses from gold would appear not only at the contact site but also systematically.

Animals↗
PubMed2000

Synthesis, crystal structure and antimicrobial activities of two isomeric gold(I) complexes with nitrogen-containing heterocycle and triphenylphosphine ligands, [Au(L)(PPh3)] (HL = pyrazole and imidazole).

Two isomeric gold(I)-triphenylphosphine complexes with nitrogen-containing heterocycles, [Au(L)(PPh3) (HL = pyrazole (1), imidazole (2)) were isolated as colorless cubic crystals for 1 and colorless plate crystals for 2, respectively. The crystal structures of 1 and 2 were determined by single-crystal X-ray diffraction. These complexes were also fully characterized by complete elemental analyses, thermogravimetric/differential thermal analyses (TG/DTA) and FT-IR in the solid state and by solution NMR (31P, 1H and 13C) spectroscopy and molecular weight measurements in acetone solution. These complexes consisted of a monomeric 2-coordinate AuNP core both in the solid state and in solution. The molecular structures of 1 and 2 were compared with those of related gold(I) complexes, [Au(1,2,3-triz)(PPh3)] (3, Htriz = triazole), [Au(1,2,4-triz)(PPh3)]2 (4) as a dimer through a gold(I)-gold(I) bond in the solid state, and [Au(tetz)(PPh3)] (5, Htetz = tetrazole). Selective and effective antimicrobial activities against two gram-positive bacteria (B. subtilis, S. aureus) and modest activities against one yeast (C. albicans) found in these gold(I) complexes 1-4 are noteworthy, in contrast to poor activities observed in the corresponding silver(I) complexes.

Anti-Bacterial Agents↗
PubMed1986

A controlled study of a 'new' ventilating tube. The gold standard?

A prospective controlled study of a gold ventilation tube was undertaken in which a gold tube was inserted in one ear of 88 children and a Paparella silicone tube was inserted in the contralateral ear as a control. The tubes were evaluated with respect to length of time of intubation, episodes of otorrhea and early occlusion. Sixty-three patients were available for a follow-up of at least 6 months. The gold tube remained in place for a shorter period of time when compared to the control tube. Episodes of otorrhea were infrequent and there was not any significant difference between the two types of tubes. The gold tube became occluded more frequently than the control tube. Since the gold tube is approximately 3 times more expensive and has no proven advantages over our conventional silicone tube, we have no reason to recommend its use in preference to any other available tube.

Acute Disease↗
PubMed2004

Effect of heat treatments on machinability of gold alloy with age-hardenability at intraoral temperature.

OBJECTIVE: This study investigated the effect of heat treatment on the machinability of heat-treated cast gold alloy with age-hardenability at intraoral temperature using a handpiece engine with SiC wheels and an air-turbine handpiece with carbide burs and diamond points. METHODS: Cast gold alloy specimens underwent various heat treatments [As-cast (AC); Solution treatment (ST); High-temperature aging (HA), Intraoral aging (IA)] before machinability testing. The machinability test was conducted at a constant machining force of 0.784N. The three circumferential speeds used for the handpiece engine were 500, 1,000 and 1,500 m/min. The machinability index (M-index) was determined as the amount of metal removed by machining (volume loss, mm(3)). The results were analyzed by ANOVA and Scheffé's test. RESULTS: When an air-turbine handpiece was used, there was no difference in the M-index of the gold alloy among the heat treatments. The air-turbine carbide burs showed significantly (p<0.05) higher M-indexes than the diamond points after any heat treatments. With the SiC wheels, increasing the circumferential speed increased the M-index values for each heat treatment. The specimens heat-treated with AC, HA and IA had similar M-indexes at the lower speeds (500 and 1,000 m/min). The ST specimens exhibited the lowest M-index at the lower speeds. However, at the highest speed (1,500 m/min), there were no significant differences in the M-indexes among the heat treatments except for HA, which showed the highest M-index. CONCLUSION: There was no effect of heat treatment on the machinability of the gold alloy using the air-turbine handpiece. The heat treatments had a small effect on the M-index of the gold alloy machined with a SiC wheel for a handpiece engine.

Body Temperature↗