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PubMed1989

Application of a silver-binding assay to the determination of protein in cerebrospinal fluid.

We evaluated a silver-binding assay for use in measuring total protein in cerebrospinal fluid. The advantage of this procedure over other methods is that, because of its sensitivity, it requires only a 0.5-microL sample. The procedure, which takes approximately 40 min to complete, involves dilution of 0.5-microL samples to 1 mL with distilled water containing sodium dodecyl sulfate, followed by addition of glutaraldehyde and an ammoniacal silver solution. After color development for 30 min, the reaction is terminated with sodium thiosulfate and the absorbance is measured at 420 nm. This assay displayed within-run and day-to-day precision (CV) of 3.1% to 13% over the range of 210 to 1370 mg/L. It showed substantially less protein-to-protein variation than the Coomassie Blue dye-binding procedure when tested with albumin, globulin, and transferrin. It also yielded an accurate estimation of hemoglobin. Moreover, preliminary studies suggested that it was capable of quantifying immunoglobulin light chains and glycoproteins. In a study of 54 human cerebrospinal fluid samples, results of the silver-binding assay corresponded more closely with those obtained with a rate biuret assay (intraclass correlation coefficient = 0.91) than did either the dye-binding or classical Lowry methods.

Albumins↗
PubMed2004

Detoxification mechanism of heavy metals in marine mammals and seabirds: interaction of selenium with mercury, silver, copper, zinc, and cadmium in liver.

Subcellular distribution of mercury, selenium, silver, copper, zinc, and cadmium was determined in the liver of northern fur seals (Callorhinus ursinus), black-footed albatrosses (Diomedea nigripes), and Dall's porpoises (Phocoenoides dalli). Mercury, selenium, and silver were preferentially accumulated in nuclear, lysosomal, and mitochondrial fraction with an increase in their hepatic concentrations, whereas copper, zinc, and cadmium were accumulated mainly in cytosol with an increase in the hepatic concentrations for all three species. To gain insight into the existing state of the metals, they were extracted with four extractants--sodium dodecylsulfate (SDS); 2-mercaptoethanol; 2-mercaptoethanol + guanidinium thiocyanate; and copper sulfate (CuSO4)--at several concentrations from nuclear, lysosomal, and mitochondrial fraction in liver from a specimen of northern fur seal. Extraction efficiencies of the metals for 2-mercaptoethanol + guanidinium thiocyanate and CuSO4 were much higher than those for SDS and 2-mercaptoethanol. Also, for all individuals of the three species, metals were extracted by the three extractants--2% SDS; 0.25 mol/L 2-mercaptoethanol + 5 mol/L guanidinium thiocyanate; and 0.1 mol/L CuSO4--from nuclear, lysosomal, and mitochondrial fraction of liver. In the northern fur seals with higher concentration of mercury, the molar ratio of selenium to mercury approached unity in the nonextractable fraction of 0.25 mol/L 2-mercaptoethanol + 5 mol/L guanidinium thiocyanate, suggesting the possible formation of mercuric selenide (HgSe) with increasing hepatic concentration. Because the nonextractable content of mercury and its distribution were larger for black-footed albatross than those for the other two species, it was suggested that the black-footed albatross has a stronger ability to form a stable compound(s) of mercury in the liver. It is notable that the existing state of silver was similar to that of mercury as judged by their subcellular distribution and the extraction tests, suggesting that silver also interacted with selenium in the liver of marine animals used in this study.

Animals↗
PubMed1991

Nucleolar organizer regions in malignant melanoma and melanocytic nevi. Comparison of two counting methods.

Using a silver staining technique, we studied nucleolar organizer regions (AgNOR) in paraffin sections of junctional nevi, compound nevi, intradermal nevi, blue nevi, dysplastic nevi, Spitz nevi, lentigo maligna, malignant melanomas in nevus, superficial spreading melanomas, and nodular melanomas. Two methods of counting black dots within nuclei were employed. One method was to count the discrete black dots within the nuclei, including the tiny black dots seen within the nucleolus; the second method did not take into account the subsidiary cluster of tiny black dots seen within the nucleolus, instead treating these dots as a single structure. Whichever method we used, a significant difference was found between the pooled mean AgNOR numbers for benign and malignant lesions. We found an overlap, however, between benign, in particular Spitz and dysplastic nevi, and malignant lesions when considering individual counts of AgNOR using both methods. We conclude that studying AgNOR does not seem to be a useful technique to differentiate Spitz and dysplastic nevi from malignant melanomas.

Diagnosis, Differential↗
PubMed1999

Initiation and elongation of lateral roots in Lactuca sativa.

Lactuca sativa cv. Baijianye seedlings do not normally produce lateral roots, but removal of the root tip or application of auxin, especially indole-butyric acid, triggered the formation of lateral roots. Primordia initiated within 9 h and were fully developed after 24 h by activating the pericycle cells opposite the xylem pole. The pericycle cells divided asymmetrically into short and long cells. The short cells divided further to form primordia. The effect of root tip removal and auxin application was reversed by 6-benzylaminopurine at concentrations >10(-8) M. The cytokinin oxidase inhibitor N1-(2chloro4pyridyl)-N2-phenylurea also suppressed auxin-induced lateral rooting. The elongation of primary roots was promoted by L-alpha-(2-aminoethoxyvinyl) glycine and silver ions, but only the latter enhanced elongation of lateral roots. The data indicate that the induction of lateral roots is controlled by basipetally moving cytokinin and acropetally moving auxin. Lateral roots appear to not produce ethylene.

Adenine↗
PubMed2005

Structure determination of homoleptic AuI, AgI, and CuI aryl/alkylethynyl coordination polymers by X-ray powder diffraction.

This article describes the structure determination of five homoleptic d(10) metal-aryl/alkylacetylides [RC triple bond CM] (M=Cu, R=tBu 1, nPr 2, Ph 3; R=Ph, M=Ag 4; Au 5) by using X-ray single-crystal and powder diffraction. Complex 1.C6H6 reveals an unusual Cu20 catenane cluster structure that has various types of tBuC triple bond C-->Cu coordination modes. By using this single-crystal structure as a starting model for subsequent Rietveld refinement of X-ray powder diffraction data, the structure of the powder synthesized from CuI and tBuC triple bond CH was found to have the same structure as 1. Complex 2 has an extended sheet structure consisting of discrete zig-zag Cu4 subunits connected through bridging nPrC triple bond C groups. Complex 3 forms an infinite chain structure with extended Cu-Cu ladders (Cu-Cu=2.49(4)-2.83(2) A). The silver(I) congener 4 is iso-structural to 3 (average Ag-Ag distance 3.11 A), whereas the gold(I) analogue 5 forms a Au...Au honeycomb network with PhC triple bond C pillars (Au-Au=2.98(1)-3.26(1) A). Solid-state properties including photoluminescence, nu(C triple bond C) stretching frequencies and thermal stability of these polymeric systems are discussed in the context of the determined structures.

Acetylene↗
PubMedDate not supplied

Norfloxacin and silver-norfloxacin as topical antimicrobial agents: results of in vitro susceptibility testing against bacteria and Candida sp isolated from burn patients.

Because of its broad spectrum of antibacterial action, Norfloxacin, a new quinoline carboxylic acid compound, was formulated in a topical cream base and tested for activity against a variety of bacteria and yeasts isolated from burn patients. An in vitro agar well diffusion topical susceptibility test was used. The silver salt of Norfloxacin also was tested. For comparative purposes, commonly used topical antibacterial and antifungal preparations were examined as well. No bacterial isolate tested was resistant to the action of Norfloxacin or that of silver-Norfloxacin. Furthermore, all Candida organisms assayed were susceptible to silver-Norfloxacin. All other topical agents tested had some strains of bacteria or yeast, or both, that were resistant to their action. Therefore, Norfloxacin and, particularly, silver-Norfloxacin, warrant further development as topical anti-infective agents for use in treating burn patients.

Anti-Infective Agents↗
PubMed1984

Cellular and subcellular demonstration of mercury in situ by modified sulfide-silver technique and photoemulsion histochemistry.

Young adult C57BL/6J mice were injected with 4.0 mg/kg body wt of methylmercuric chloride (MMC) for 3 consecutive days for a total of 12.0 mg/kg. Control animals received physiological saline in place of MMC. One week following the last injection, the animals were sacrificed. Representative tissue blocks and sections from the brain, kidney, and liver were subjected to a modified sulfide-silver technique (SST) and the photoemulsion histochemical method. The results show that both of these techniques demonstrate consistent and distinct localization of mercury (Hg) gains in cells and subcellular organelles. These methods are based on the principle that Hg compounds react strongly with sulfhydryl groups in tissues and cells to form Hg-sulfides and also on the affinity of Hg and silver to form an amalgam when placed in a physical developer or photographic emulsion. Thus Hg in cells is demonstrable without prior treatment of sulfide solution. The methods are simple and reliable when used with proper controls. Specific localization of Hg in cells in situ without the use of radioactive material and without disruption of anatomical relationships provided by these methods offers distinct advantages over other methods of Hg determination. Thus it would be possible to conduct a retrospective or prospective study of human autopsy materials and also would allow direct correlation of Hg deposition with pathological changes in cells and subcellular organelles.

Animals↗
PubMed2001

Si extraction from silica in a basic polychalcogenide flux. Stabilization of Ba4SiSb2Se11, a novel mixed selenosilicate/selenoantimonate with a polar structure.

An unusual compound, Ba4SiSb2Se11, was discovered from a reaction of Ba/Th/Sb/Se. It is assumed that Si was extracted from the silica reaction tube. It forms as silver needlelike crystals in the polar space group Cmc2(1) with a = 9.3981(3) A, b = 25.7192(7) A, c = 8.7748(3) A, and Z = 4. A rational synthesis has been devised at 600 degrees C. The compound is composed of Ba2+ ions stabilized between infinite one-dimensional [SiSb2Se11]8- chains running parallel to the a axis. Each chain is composed of a [SbSe2]- infinity backbone with [SiSe4]4- tetrahedra chelating every other Sb atom from the same side of the backbone. The V-shaped triselenide groups, (Se3)2-, are attached to the rest of the Sb atoms in the chain through one of their terminal Se atoms. The compound has a band gap of 1.43 eV. The Raman spectrum shows a broad shift at 247 cm-1 and a shoulder around 234 cm-1, which are related to the Se-Se vibration of the triselenide groups and/or the Si-Se vibrations of the [SiSe4]4- groups. The compound decomposes at 522 degrees C.

Journal Article↗
PubMed2003

[Improvement of the silver-stain and it's test and verification in comet assay].

In order to find out a better silver-stain method so that to replace EB-stain in comet assay, the selected silver-stain methods were tested via determining fixing, coloring, displaying, fading, and testing the pigmentation temperature. When it was optimized, it was used in testing the DNA damage by two types of compounds, namely potassium dichromate (K2Cr2O7) and 1,2-dichloroethane (DCE) to two kind of cells, namely spleen cells of mouse and human outer lymphocyte, and, the sensitivity was compared with that of EB-stain. The results showed that the modified silver-stain was the first successful non-fluorescent dye method in comet assay that can dye clearly, permanently, with a light background and good contrast, and it was easy to manipulate and it was cheap. The stain can detect the DNA damage by compounds to cells and has the same positive threshold like that in EB-stain. It can even dye the small fragments that EB-stain cannot. It is suggested that in comet assay, the silver-stain can substitute the EB-stain completely.

Comet Assay↗
PubMed1994

The localization of mercury and metallothionein in the cerebellum of rats experimentally exposed to methylmercury.

Rats were dosed with methylmercuric chloride, either by gastric gavage (5 x 10 mg kg-1 body weight over a 15-day period), or in their drinking water (20 mg methylmercuric chloride l-1 for 14 or 42 days). Localization of mercury within the cerebellum was performed with a silver physical development technique, and metallothionein with dinitrophenyl hapten-sandwich immunohistochemistry. Mercury was detected in structurally undamaged Purkinje neurones and adjacent Bergmann glial cells; no mercury was detected in granule cells even though these small cells nearest the Purkinje layer had a high incidence of pyknotic nuclei. In general, metallothionein was detected mainly in Bergmann glial cells, Purkinje cells, astrocytes and glial cells of white matter; no metallothionein was detected in granule cells. We hypothesized that the resistance of Purkinje cells to methylmercuric chloride reflects their ability to transform organic mercurials to inorganic mercury that, in turn, induces the synthesis of radical-scavenging metallothionein molecules.

Administration, Oral↗
PubMed1999

Synthesis and Characterization of Functionalized N,N'-Diphenylformamidinate Silver(I) Dimers: Solid-State Structures and Solution Properties.

Functionalized N,N'-diphenylformamidines and their deprotonated silver(I) complexes have been synthesized: silver(I) N,N'-di(4-alkyl)phenylformamidinate (alkyl = methyl, ethyl, n-butyl, and n-hexadecyl) 1-4; silver(I) N,N'-di(4-trifluoromethyl)phenylformamidinate 5, silver(I) N,N'-di(3-methoxy)phenylformamidinate 6, silver(I) N,N'-di(3-methylmercapto)phenylformamidinate 7, silver(I) N,N'-di(2-methoxy)phenylformamidinate 8, silver(I) N,N'-di(2-methylmercapto)phenylformamidinate 9. The effects of increasing the coordination number of the silver(I) centers by donor substituents on the phenyl groups have been investigated by solution and solid-state studies. Variable-temperature proton NMR (223-303 K) for 1-4 shows coupling between the proton attached to the amidinate carbon and the (107/109)Ag centers at room temperature which is unaffected by cooling (2). For the four-coordinate complexes, 8 and 9, such coupling is only observed on cooling. Molecular weight measurements recorded in solution by vapor pressure osmometry at 310 K show some aggregation to higher molecular weight species than simple dimers for 1-4 and 6, but 8 and 9 exist as discrete dimeric species. Measurement of thermal stability shows the expected increase in stability with increasing coordination number. Compounds 8 and 9 were structurally characterized by X-ray methods. Both show four-coordinate silver dimers bridged by two amidinate ligands with additional longer interactions with the ether oxygens or thioether sulfurs.

Journal Article↗
PubMed1998

Detection of membrane-bound enzymes in cells using immunoassay and Raman microspectroscopy.

The method of surface-enhanced Raman microspectroscopy was developed for direct detection of membrane-bound enzymes in cells. Cells were cultured, fixed, and incubated with specific primary antibodies and their corresponding labeled secondary antibodies, and surface-enhanced Raman scattering (SERS) was detected directly in the wells of a multiwell plate. First, specific primary antibodies were separately bound to enzymes in cells. Then, the peroxidase-labeled secondary antibodies were added to bind these primary antibodies. Peroxidase substrates, o-phenylenediamine and hydrogen peroxide, were added and reacted for 15 min at room temperature to form azoaniline, a compound with strong Raman scattering. Then, Raman scattering of this enzymatic product was enhanced by silver colloids. Samples were excited with a He/Ne laser at 632.8 nm and SERS was detected by a CCD camera. The SERS spectrum of this product showed an intense peak at 1370 cm-1 and its intensity was used for assessment of cellular enzymes. The observed amount of enzyme was normalized to protein content in each well. The method was successfully used to detect prostaglandin H synthase-1 and -2 (PGHS-1 and -2) in normal human hepatocytes and human hepatocellular carcinoma (HepG2) cells. The detection limit of these PGHS enzymes by this method was about 0.1 pg per well. An immunohistochemical staining was also used to detect the expression of both PGHS isozymes in these cells.

Cell Membrane↗
PubMed2000

Adsorption and Chemical Modification of Phenols on a Silver Surface.

The adsorption of phenols of different natures on silver colloidal particles is studied here by surface-enhanced Raman spectroscopy (SERS). The studied compounds can be classified in three groups: (a) cinnamic acic derivatives: caffeic and isoferulic acids; (b) catechol; and (c) the phenols derived from benzoic acid: m- and p-hydroxybenzoic acids and salicylic, vanillic, and gallic acids. The interest of these compounds lies in the fact that they are naturally occurring molecules with significant importance in relation to plant metabolism, soil chemistry, and vegetal food stability. In addition, many of these compounds have antioxidant properties derived from their high affinity toward atmospheric oxygen. They exhibit high reactivity that may be enhanced in the presence of a metal surface such as those employed for SERS spectroscopy. From the SERS results it can be deduced that a clear chemical change of caffeic and gallic acid and catechol occurred. The chemical modification consists mainly of polymerization connected to existence in the molecule of o-diphenol moieties. In the case of m-hydroxybenzoic acid the chemical change may occur at low pH at which a reorientation of the molecule on the surface takes place, while in the o-hydroxybenzoic acid the only chemical change seems to be the internal H bond breakdown induced by the complexation with the metal. Finally, isoferulic and p-hydroxybenzoic acids do not show any chemical modification upon adsorption on the metal, which takes place through the carboxylate group adopting the molecule a standing up orientation. The case of vanillic acid is not so clear, although possible chemical modification is also possible for this adsorbate. From the results found in this work it can be inferred that the factors influencing possible chemical modification are the chemical structure of the adsorbate and its orientation and interaction with the surface. Copyright 2000 Academic Press.

Journal Article↗
PubMed1988

Synthesis of antigenic determinants of the Brucella A antigen, utilizing methyl 4-azido-4,6-dideoxy-alpha-D-mannopyranoside efficiently derived from D-mannose.

A strategy for the synthesis of Brucella O-antigenic determinants containing 2-linked 4,6-dideoxy-4-formamido-alpha-D-mannopyranosyl residues is described. The approach adopted also permits the N-acyl moiety to be varied. A high-yield synthesis of methyl 4-azido-4,6-dideoxy-alpha-D-mannopyranoside from D-mannose on the 10-20-g scale provided the key intermediate. Regioselective acetylation of 7 gave the 2-acetate 8, which, on treatment with benzyl trichloroacetimidate, provided methyl 2-O-acetyl-4-azido-3-O-benzyl-4,6-dideoxy-alpha-D-mannopyranoside. This compound served as a common precursor to the glycosyl donor 12 and acceptor 10 molecules. Silver trifluoromethanesulphonate-promoted glycosylation of 10 by 12 gave a disaccharide derivative, hydrogenolysis of which gave methyl 4-amino-2-O-(4-amino-4,6-dideoxy-alpha-D-mannopyranosyl)-4,6-dideoxy- alpha-D-mannopyranoside from which the N-formyl and N-acetyl derivatives were obtained. Deacetylation of 13 followed by glycosylation with 12 gave a trisaccharide derivative. The N-formylated disaccharide 17 inhibited the binding of Brucella O-polysaccharide to Brucella-specific monoclonal antibodies.

Antigens, Bacterial↗
PubMed1992

Neurons of origin of zinc-containing pathways and the distribution of zinc-containing boutons in the hippocampal region of the rat.

Recent methods allow the study of neurons that contain zinc in synaptic vesicles of their boutons (Timm-stainable boutons) by the intravital precipitation (local or throughout the CNS) of the vesicular zinc with selenium compounds and its subsequent retrograde transport to the parent neurons, where the precipitate can be silver enhanced. The present study is a description of the distribution of zinc-containing neurons, their possible connections and their terminal fields within the hippocampal region of the rat. Problems inherent to the methods are addressed. Finally, based on the results and a review of literature, the possible function of zinc in the hippocampal region is considered. Neurons which contain silver-enhanced precipitates were observed in layers II, V and VI of the lateral entorhinal area and in layers V and VI of the medial entorhinal area. In the parasubiculum, labeled cells were seen in layer II/III of the parasubiculum a and in layer V. Labeled cells in the presubiculum were concentrated in layers III and V, in the hippocampal pyramidal cell layer and the dentate granule cell layer, but neurons containing precipitates were largely absent from the subiculum. Zinc-containing axonal boutons defined subpopulations within principal hippocampal neuron populations. Within layer II of the lateral entorhinal cortex and the pyramidal cell layer for regio inferior deeply situated neurons were labeled, whereas superficially placed pyramidal cells were labeled in regio superior. The neuropil staining described in the present study corresponded to that found in earlier studies. However, glial and vascular staining or unspecific background were largely absent, and the neuropil staining could unequivocally be identified light microscopically. Methodological problems are most prominently reflected in unstained mossy fibers in some animals. Based on series from animals treated with decreasing doses of sodium selenite and increased survival times, this problem can be related to small amounts of circulating reactive selenium and a competition of zinc compartments (vesicles) for the selenium. Staining will fail where the competition prevents individual compartments from reaching a threshold amount of zinc precipitate for silver amplification. A guide to evaluate histological material is provided. The distribution of zinc-containing boutons and their cells of origin indicate that zinc-containing and zinc-negative projections are not organized as parallel pathways. The mossy fibers provide an example of a pure zinc-containing pathway. Projections from regio superior to the dorsal presubiculum are likely to be zinc-negative while projections from the same area to the subiculum are zinc-containing.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗