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Aqueous-organic phase-transfer of highly stable gold, silver, and platinum nanoparticles and new route for fabrication of gold nanofilms at the oil/water interface and on solid supports.

A simple but effective aqueous-organic phase-transfer method for gold, silver, and platinum nanoparticles was developed on the basis of the decrease of the PVP's solubility in water with the temperature increase. The present method is superior in the transfer efficiency of highly stable nanoparticles to the common phase-transfer methods. The gold, silver, and platinum nanoparticles transferred to the 1-butanol phase dispersed well, especially silver and platinum particles almost kept the previous particle size. Electrochemical synthesis of gold nanoparticles in an oil-water system was achieved by controlling the reaction temperature at 80 degrees C, which provides great conveniences for collecting metal particles at the oil/water interface and especially for fabricating dense metal nanoparticle films. A technique to fabricate gold nanofilms on solid supports was also established. The shapes and sizes of gold nanoparticles as the building blocks may be controllable through changing reaction conditions.

Journal Article↗

Redox and ligand exchange reactions of potential gold(I) and gold(III)-cyanide metabolites under biomimetic conditions.

Biomimetic pathways for the oxidation of [Au(CN)(2)](-), a gold metabolite, and further cyanation of the gold(III) products to form Au(CN)(4)(-) were investigated using 13C NMR and UV-Visible spectroscopic methods. Hypochlorite ion, an oxidant released during the oxidative burst of immune cells, was employed. The reaction generates mixed dicyanoaurate(III) complexes, trans-[Au(CN)(2)X(2)](-), where X(-) represents equilibrating hydroxide and chloride ligands, and establishes the chemical feasibility of dicyanoaurate oxidation by OCl(-) to gold(III) species. This oxidation reaction suggests a new procedure for synthesis of H[Au(CN)(2)Cl(2)]. Reaction of trans-[Au(CN)(2)X(2)](-) (X(-)=Cl(-) and Br(-)) or [AuCl(4)](-) with HCN in aqueous solution at pH 7.4 leads directly to [Au(CN)(4)](-) without detection of the anticipated [Au(CN)(x)X(4-x)](-)intermediates, which is attributed to the cis- and trans-accelerating effects of the cyanides. The reduction of [Au(CN)(4)](-) by glutathione and other thiols is a complex, pH-dependent process that proceeds through two intermediates and ultimately generates [Au(CN)(2)](-). These studies provide further insight into the possible mechanisms of an immunogenically generated gold(I)/gold(III) redox cycle in vivo.

Cyanates↗

Use of the protein A-gold immunocytochemical and enzyme-gold cytochemical techniques in studies of vitellogenesis.

Vitellogenesis in the frog hepatocyte was investigated by applying the protein A-gold immunocytochemical and RNase-gold cytochemical techniques in conjunction with morphometric and biochemical analyses. The morphometric studies demonstrated that the surface density of rough endoplasmic reticulum (RER) and nucleolar size increased more than fourfold and 1.25-fold, respectively, while the nuclear size and the mitochondrial compartment size remained constant following estrogen treatment. Concurrently, liver RNA concentration increased 2.5-fold while protein and DNA concentrations did not change. In addition, total plasma protein more than doubled, with vitellogenin accounting for 40% of the final volume. The secretory proteins vitellogenin and protein-RcX (a nonvitellogenin, estrogen-induced plasma protein of unknown function, found in the plasma of Rana catesbeiana) were detected immunocytochemically in the RER, Golgi apparatus, and secretory granules in hepatocytes only of estrogen-treated frogs. Lysosomes also were labeled. These observations established that protein-RcX was synthesized and secreted by the hepatocyte in parallel with vitellogenin and that both of these export proteins were confined to the secretory pathway and lysosomes. Quantitation of labeling density indicated that the concentration of vitellogenin increased as it progressed along the secretory vector. Albumin was detected immunocytochemically also within these same hepatocyte entities from both untreated and treated animals. In the untreated animals, albumin concentration also increased progressively along the secretory vector. A marked alteration of albumin processing was observed following estrogen treatment. While albumin concentration in the RER was unchanged, its concentrations within the Golgi apparatus and secretory granules were lower than those observed in the RER or in counterpart compartments under control conditions. RNase-gold cytochemistry for total RNA demonstrated a 1.5-fold increase in labeling density over the nucleolus but no change in RER labeling following estrogen treatment. These labeling data, in combination with the morphometric data, suggest an increase of approximately 80% in the total amount of RNA in the nucleolus and 430% in the RER in response to estrogen. This review thus illustrates the significant contributions which can be made by gold-probe techniques, alone or in combination with morphometric and biochemical techniques, to investigations of the intracellular processing of secretory proteins.

Animals↗

"Thiocyanate gold": small (2-3 nm) colloidal gold for affinity cytochemical labeling in electron microscopy.

Reduction of HAuCl4 by NaSCN or KSCN produces colloidal gold particles of 2.6 nm in diameter and homogeneous in size (coefficient of variation approximately 15%). The AuSCN sol forms protein-gold complexes. The amount of protein required to form an AuSCN-protein complex is best determined in the electron microscope, where serial dilutions of protein with gold sol are inspected for the presence of aggregates. By immuno-electron microscopy SCN-gold complexed to protein A is active and visible as is shown by revealing alpha-amylase in rat pancreatic acinar cells.

Animals↗

Chitinase-gold complex used to localize chitin ultrastructurally in tomato root cells infected by Fusarium oxysporum f. sp. radicis-lycopersici, compared with a chitin specific gold-conjugated lectin.

A cytochemical technique for the ultrastructural localization of chitin in tomato root cells infected by Fusarium oxysporum f. sp. radicis-lycopersici is reported. Chitinase was complexed to colloidal gold and thin sections were incubated with the enzyme-gold complex. This technique yielded a more uniform distribution of gold particles over the fungus wall, compared to that obtained with the lectin-gold technique. Both techniques revealed no labelling of the fungus cytoplasm, except for organelles resembling Woronin bodies. No significant labelling of either healthy or infected root cells was seen except for the secondary walls of vessels and, occasionally, that of adjoining parenchyma cells. The importance of this technique in studying the development of the pathogen within host cells is discussed.

Chitinases↗

The preparation of protein A-gold complexes with 3 nm and 15nm gold particles and their use in labelling multiple antigens on ultra-thin sections.

The preparation of a protein A-gold complex (pAg3) using 3 nm gold particles and its application for labelling of intracellular antigens on thin sections is reported. The 3 nm gold particle is the smallest metal particle currently available for cytochemistry and permits a higher resolution of the pAg technique. Furthermore, it can be used in double labelling experiments in conjunction with a pAg complex prepared from 15nm gold particles. For double labelling, the pAg3 complex must be used for staining of the first antigen since otherwise a non-specific co-labelling of the two pAg complexes results.

Animals↗

Silver enhancement of lectin-gold and enzyme-gold cytochemical labelling of eggs of the nematode Onchocerca gibsoni.

Wheat germ agglutinin-gold and chitinase-gold complexes were used to demonstrate the presence of chitin on the surfaces of eggs of the animal parasitic nematode Onchocerca gibsoni. The gold complexes were enhanced by silver intensification and examined by light microscopy (LM), transmission electron microscopy (TEM) and scanning electron microscopy (SEM). Distinctive labelling of the egg surfaces was obtained with both probes in all three microscope modes. The results indicate that the small colloidal gold markers (3-10 nm) commonly used for high resolution TEM studies may be silver enhanced and also used for sensitive LM and SEM studies.

Animals↗

Subarachnoidal macrophages share a common epitope with resident non-cerebral macrophages and show receptor-mediated endocytosis of albumin-gold and IgG-gold complexes.

Cerebral macrophages are supposed to exploit a pivotal role in scavenger functions of the central nervous system. We have examined the in vivo uptake of serum albumin and IgG conjugated with colloidal gold by subarachnoidal macrophages. These serum-borne proteins are endocytosed by receptor-mediated endocytosis by varying kinetics. Albumin-gold conjugates were found to be associated to a significant amount with coated pits and coated vesicles 1 min after superfusion of the cerebral surface. Within 25 min the major fraction of albumin-gold was transferred to the lysosomal compartment. IgG-conjugates revealed a less pronounced uptake. The uptake of albumin-gold could be competed by saturating of the 'receptor sites' with free albumin. It is suggested that the described receptor-mediated uptake of serum-borne proteins by subarachnoidal macrophages serves a cleansing function following blood-brain barrier disruption during acute or subacute inflammatory reactions.

Animals↗

The fabrication of a colloidal gold-carbon nanotubes composite film on a gold electrode and its application for the determination of cytochrome c.

Colloid Au (Au(nano)) with a diameter of about 20 nm was prepared and used in combination with the multi-wall carbon nanotubes (MWNTs) to modify a gold electrode. Dihexadecylphosphate (DHP) dispersed in Au(nano) aqueous solution was used to solubilize MWNTs. Deposition of Au(nano) on MWNTs was realized as illustrated by TEM micrographs. The DHP formed a network that connected Au(nano) and MWNTs to the gold electrode surface. The Au(nano)-MWNTs-DHP composite film on the gold electrode surface was characterized by electrochemical impedance spectroscopy (EIS) and cyclic voltammmetry (CV). The composite film modified gold electrode was used to detect cytochrome c and a pair of well-defined redox waves was obtained. It was found that the composite film promoted the redox of horse heart cytochrome c and its effect was developed for the determination of cytochrome c.

Biosensing Techniques↗

Two-dimensional arrangement of a functional protein by cysteine-gold interaction: enzyme activity and characterization of a protein monolayer on a gold substrate.

We have characterized the functional protein, myosin subfragment 1 (S1), attached to a gold substrate by the sulfhydryl groups of cysteine in proteins. The amino groups of the regulatory light chain (RLC) isolated from myosin were labeled with a radioisotope (125I), and the labeled RLC was incorporated into S1 from which the RLC had been removed. The radiation from 125I showed that S1 molecules had attached to the gold and, through the interference effect of the monochromatic radiation from 125I, provided information about the position of labeled RLC sites in the S1 monolayer. The interference fringes showed that the RLC was located close to the gold surface and that all of the adsorbed S1 molecules had the same orientation. We confirmed that the motor function of S1 on the gold surface is maintained by observing sliding movement at low ionic strength and by observing the detachment at high ionic strength of fluorescent actin filaments in the presence of ATP. We also found that the adsorbed S1 molecules were not removed from the Au surface by a reducing agent. Thus the Au-S bond is more stable than the S-S bond.

Actins↗

Gold finder: a computer method for fast automatic double gold labeling detection, counting, and color overlay in electron microscopic images.

This work presents a computerized method to identify, detect, evaluate, and, by colored overlay, present gold particle pairs in electron microscopy (EM), even in wide-field views. Double gold immunolabeled specimens were analyzed in a LEO 912 electron microscope equipped with a 2k x 2k-pixel slow-scan cooled CCD camera connected to a computer with analySIS 3.1 PRO image processing software. The acquisition of a high-resolution and high-dynamic-range image by the camera allowed correct segmentation of the gold particles, separating them from other cell structures and from the substrate. Particle identification was performed by a classification module designed by us. Based on shape and size, the computer recognized the group of small particles and classified them as either singular or clustered and differentiated these from the single bigger type. The final image shows the particle types separated and colored, and indicates the total number of objects encountered in the specific region of interest. Moreover, a montage tool allowed us to obtain final representative images of large microscopic fields, which on analysis by the Gold Finder module provided information on the distribution and localization of antigens comparable to that provided by the wide-field light microscope images.

Animals↗

Ligand density effect on biorecognition by PEGylated gold nanoparticles: regulated interaction of RCA120 lectin with lactose installed to the distal end of tethered PEG strands on gold surface.

PEGylated gold nanoparticles (diameter: 20 nm) possessing various functionalities of lactose ligand on the distal end of tethered PEG ranging from 0 to 65% were prepared to explore the effect of ligand density of the nanoparticles on their lectin binding property. UV-visible spectra of the aqueous solution of the nanoparticles revealed that the strong steric stabilization property of the PEG layer lends the nanoparticles high dispersion stability even under the physiological salt concentration (ionic strength, I = 0.15 M). The number of PEG strands on a single particle was determined to be 520 from thermogravimetric analysis (TGA). Scanning electron microscopy (SEM) observation under controlled acceleration voltage revealed the thickness of the PEG layer on the nanoparticle to be approximately 7 nm. The area occupied by a single lactose molecule on the surface of PEGylated gold nanoparticles was then calculated based on TGA and SEM results and was varied in the range of 10-34 nm2 depending on the lactose functionality (65 approximately 20%). PEGylated gold nanoparticles with 40% and 65% lactose functionality showed a selective and time-dependent aggregation in phosphate buffer with the addition of Ricinus communis agglutinin (RCA120) lectin, a bivalent galactose-specific protein. The aggregates can be completely redispersed by adding an excess amount of galactose. Time-lapse monitoring of UV-visible spectra at 600-750 nm revealed that the aggregation of PEGylated gold nanoparticles was accelerated with an increase in both RCA120 concentration in the solution and the lactose density of the nanoparticles. Furthermore, the sensitivity of lectin detection could be controlled by the regulation of lactose density on the particle surface. Interestingly, there was a critical lactose density (>20%) observed to induce detectable particle aggregation, indicating that the interaction between the particles is triggered by the multimolecular bridging via lectin molecules.

Dimerization↗

Mercury poisoning: prevalence, knowledge and frequency of gold panning and doing retort among alluvial gold panners in Chiweshe and Tafuna communal lands in Zimbabwe.

OBJECTIVES: To estimate the prevalence of mercury poisoning, to estimate the knowledge level that mercury can be a poison, and to establish the frequency of gold panning and doing retorts. DESIGN: Cross sectional study. SETTING: Chiweshe and Tafuna communal lands. SUBJECTS: Gold panners. MAIN OUTCOME MEASURE: Mercury levels in blood and urine. RESULTS: Totals of 23 respondents from Chiweshe and 43 respondents from Tafuna were recruited. Four out of 43 respondents in Tafuna and seven out of 23 respondents in Chiweshe had levels of mercury greater than 0.05 mg/L in blood (p = 0.040). Out of 43 respondents in Tafuna, four (9.3%) had levels of mercury of more than 0.01 mg/L in urine. Totals of 18 out of 37 and seven out of 22 respondents from Tafuna and Chiweshe, respectively, did not know that mercury could be a poison. Altogether, 35 (56.5%) out of 62 respondents were full time gold panners. Significantly more respondents in Chiweshe (14/19) than in Tafuna (8/29) did less than four retorts per month (p = 0.005). Respondents who did four or more retorts per month were 3.21 (95% CI 1.06 to 9.72) times more likely to have had raised levels of mercury in their blood compared with persons who did less than four retorts per month. CONCLUSION: Mercury poisoning among gold panners in Chiweshe and Tafuna communal lands is of public health importance. Panners should be educated on the possibilities of mercury being a poison. A low cost and safe technology to separating mercury from the amalgam should be introduced to the panners.

Adult↗

Measurement of gold treatment effect in clinical practice: evidence for effectiveness of intramuscular gold therapy.

OBJECTIVE: To determine the extent of efficacy of intramuscular (im) gold in clinical practice. METHODS: Ninety-eight patients who received im gold treatment for at least 1 year were assessed during ordinary clinic visits over the 12 month period following gold initiation. Assessments included joint counts, erythrocyte sedimentation rate (ESR), morning stiffness, visual analog scale (VAS) patient pain, VAS patient global, grip strength, prednisone dose, hemoglobin, and Stanford Health Assessment Questionnaire (HAQ) disability index. RESULTS: All outcome measures showed significant improvements at one year by standardized pretest posttest differences (effect size) and by measurement of the area under the curve (AUC). Important improvement (at least 50% improvement) was common: joint count (63.3%), global severity (41.1%), ESR (48.2%), grip strength (42.7%) HAQ disability (45.3%), and morning stiffness (72.2%). Effect sizes were substantial, even after correcting for placebo effect and withdrawals. Overall improvement was clinically significant, amounting to the equivalent of 2 months without pain or disability. CONCLUSION: Patients receiving gold for 12 months improved significantly and importantly in all measures studied. Improvement was clinically significant, amounting to the equivalent of 2 months without pain or disability. The degree of improvement was similar to, but somewhat greater, than that seen in controlled clinical trials, and could not be explained by regression to the mean.

Arthritis, Rheumatoid↗

Syntheses of mixed-ligand tetranuclear gold(I)-nitrogen clusters by ligand exchange reactions with the dinuclear gold(I) formamidinate complex Au(2)(2,6-Me2Ph-form)2.

The reaction of the sterically crowded dinuclear gold(I) amidinate complex Au2(2,6-Me2Ph-form)2, 1, with the less bulky bidentate nitrogen ligands results in the formation of tetranuclear gold(I) complexes. When the less bulky amidinate, K(4-MePh-form), A, was reacted with 1 in a 1:1 stoichiometric ratio, crystals containing equal amounts of the tetranuclear and dinuclear gold(I) aryl formamidinates, Au4(4-MePh-form)4 and Au2(2,6-Me2Ph-form)2, where 2,6-Me2Ph-form = B, were found in the same unit cell, 2 x 2THF: space group P, a = 10.794(11) A, b = 14.392(15) A, c = 25.75(3) A, alpha = 82.564(17) degrees, beta = 85.443(18) degrees, gamma = 82.614(19) degrees. The reaction of K(4-MePh-form), A, and 1 in a 1:2 ratio (excess) produced the tetranuclear complex only, 3. The potassium salt of the exchanged bulky ligand, K(2,6-Me2Ph-form), formed as a byproduct. The reaction of the dinuclear gold(I) complex Au2(2,6-Me2Ph-form)2 with the 3,5-diphenylpyrazolate salt, K(3,5-Ph2pz), resulted in the formation of two tetranuclear mixed-ligand complexes, Au4(3,5-Ph2pz)2(2,6-Me2Ph-form)2 x 2THF, 4 x 2THF (space group P21/c, a = 11.5747(19) A, b = 25.497(4) A, c = 21.221(3) A, beta = 96.979(3) degrees) and Au4(3,5-Ph2pz)3(2,6-Me2Ph-form) x THF, 5 x THF (space group P21/c, a = 23.058(5) A, b = 14.314(3) A, c = 18.528(4) A, beta = 90.94(3) degrees. The block crystals from the tetranuclear complex, 4 x 2THF, contain mixed ligands with each pyrazolate ring facing an amidinate ring. The tetranuclear mixed ligand complex, 5 x THF, was isolated as needles with ligands alternating above and below the Au4 plane. The two tetranuclear mixed-ligand complexes emit at 490 and 530 nm, respectively, under UV excitation.

Journal Article↗

Gold(I)-Purine Interactions: Synthesis and Characterization of Cyclic and Open Chain Polynuclear Gold(I) Complexes Containing Xanthine Derivatives and Bis(phosphine) as Bridging Ligands. Crystal Structures of [Au(2)(&mgr;-HX)(&mgr;-dmpe)].3H(2)O and [Au(2)(&mgr;-TT)(&mgr;-dmpe)].H(2)O (H(3)X = Xanthine; H(2)TT = 8-Mercaptotheophylline).

The reaction of [(AuBr)(2)(&mgr;-PR(2)P(CH(2))(n)()PR(2))] (where R = CH(3) for n = 1; R = Ph for n = 1, 3, 4) with N-alkylxanthine and thioxanthine derivatives, containing two ionizable protons in close positions, yields, under basic conditions, neutral heterobridged dinuclear gold(I) ring complexes [Au(&mgr;-L)(2)(&mgr;-PR(2)P(CH(2))(n)()PR(2))], which have been investigated by means of (1)H- and (31)P-NMR and FAB spectroscopies. Crystal structures of two of these complexes are reported. [Au(2)(&mgr;-HX)(&mgr;-dmpe)].3H(2)O (1) (H(3)X = xanthine; dmpe = 1,2-bis(dimethylphosphine)ethane) crystallizes in the monoclinic space group P2(1)/n with a = 9.348(2) Å, b = 8.656(2) Å, c = 24.585(5) Å, beta = 98.24(2) degrees, Z = 4, and R = 0.040. [Au(2)(&mgr;-TT)(&mgr;-dmpe)].H(2)O (2) crystallizes in the monoclinic space group P2(1)/n with a = 10.853(4) Å, b = 14.031(6) Å, c = 13.574(5) Å, beta = 100.80(4) degrees, Z = 4, and R = 0.063. The structures of 1 and 2 are similar and consist of dinuclear nine-membered ring molecules, in which the two linear two-coordinate gold atoms are bridged on one side by a dmpe ligand and on the other side by a bidentate xanthinato dianion, with intramolecular Au.Au distances of 3.053(1) and 2.952 (2) Å, respectively. In the former, the coordination of the xanthinato ligand to the gold atoms takes place through the N3 and N9 nitrogen atoms whereas, in the latter, N7,S8-chelate coordination of the 8-thiotheophyllinato dianion occurs. The magnitude of the Au.Au separation is analyzed in terms of the twisting of the xanthine derivative ligand from the plane containing the gold(I) and phosphorus atoms. For n = 6 the steric requirements of the Au(&mgr;-dpph)Au group prevents the formation of dinuclear ring complexes and open chain complexes are obtained. Finally, when the xanthine derivatives do not contain two close active coordination sites dinuclear open chain complexes are formed.

Journal Article↗

Aqueous-organic phase transfer of gold nanoparticles and gold nanorods using an ionic liquid.

The water-immiscible ionic liquid, [C4MIM][PF6], is a solvent medium that allows complete transfer of gold nanoparticles from an aqueous phase into an organic phase. Both spherical and rod-shaped gold nanoparticles are efficiently transferred from an aqueous solution into the organic phase without requiring the use of thiols. The sizes and shapes of the gold nanoparticles were preserved during the phase-transfer process when a surfactant was added to the ionic liquid. This process offers a simple approach for obtaining solutions of differently sized and shaped gold nanoparticles in ionic liquids.

Journal Article↗

Divide and protect: capping gold nanoclusters with molecular gold-thiolate rings.

Density functional theory calculations are used to explore phosphine- and thiolate-protected gold nanoclusters, namely, Au(39)(PH(3))(14)Cl(6) and Au(38)(SCH(3))(24). For Au(38)(SCH(3))(24), a novel structural motif is predicted, consisting of ringlike (AuSCH(3))(4) units protecting a central Au(14) core. The calculated optical spectrum of this species features a large optical gap (about 1.5 eV) and a prominently peaked structure, correlating with experimental findings of "molecular-like spectra" of thiolate-protected 1.1 nm gold nanoparticles. Ligand-ligand interactions and steric effects in the ligand shell are suggested as possible driving forces toward an ordered gold core structure. A novel mechanism for ligand-exchange reactions on gold clusters is proposed.

Journal Article↗