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Acute leukopenia during topical burn therapy with silver sulfadiazine.

Leukopenia, with a mean white blood cell count of 2,680/mm3, was observed in nine patients with thermal injury early in the course of topical treatment with silver sulfadiazine. All manifested absolute neutropenia with a concomitant increase in immature band forms in the peripheral blood smear. The leukocyte counts returned to within normal limits within 48 to 72 hours of discontinuation of silver sulfadiazine therapy in four patients, and also did so in five patients in whom silver sulfadiazine therapy was continued. Leukopenia secondary to silver sulfadiazine application is currently believed to be an innocuous, self-limited phenomenon.

Acute Disease↗

Silver staining of proteins in polyacrylamide gels: increased sensitivity by a blue toning.

An image amplification of silver-stained proteins in polyacrylamide gels by a blue toning has been developed. Following the silver staining, this technique is fast, simple, requires only stable and inexpensive reagents, and its sensitivity is three- to sevenfold higher than the silver staining alone, and 300- to 700-fold higher than the Coomassie blue. When combined with a silver staining utilizing dithiothreitol (J. H. Morrissey, (1981) Anal. Biochem. 117, 307-310) this method is shown to be uniform in sensitivity for five standard proteins. The blue toning does not affect the linearity of the staining and allows the quantitation of proteins by densitometric evaluation.

Animals↗

Requirement for cysteine in the color silver staining of proteins in polyacrylamide gels.

To determine whether cysteine residues have a contribution to the mechanism of color silver staining, we silver stained sodium dodecylsulfate polyacrylamide gel electrophoresis separations of proteins which have few or no cysteines. Proteins without cysteine stained negatively (yellow against a yellow background) with silver. Proteins with one or more cysteines stained orange, red, brown, or green/gray depending on the mole percentage of cysteine and whether they contained covalently attached lipids. The colors could not be correlated with the mole percentages of cysteine of these proteins indicating that some components other than cysteine affect the staining color of cysteine-containing proteins. Silver staining of amino acids, sugars, nucleotide bases, or lipopolysaccharide dot-blotted onto nitrocellulose paper implicated adenine, lipids, the basic amino acids, and glutamine, but not sugars or other amino acids in silver/protein complexes.

Bacterial Outer Membrane Proteins↗

Comparison of diazo-coupling, formazan, and silver staining techniques for visualizing alkaline phosphatase isoenzymes after electrophoresis in homogeneous-pore and gradient-pore polyacrylamide gels.

Three techniques for visualization of alkaline phosphatase after polyacrylamide-gel electrophoresis are compared. These are diazo-dye simultaneous coupling with the substrate sodium naphthyl phosphate and 5-chloro-2-toluene diazonium chloride; formazan precipitation with the substrate 5-bromo-4-chloro-3-indolyl phosphate and 3-[4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide; and silver staining with the substrate sodium glycerophosphate. Each staining technique was tested with gradient-pore and homogeneous-pore acrylamide-gel electrophoresis. The main factors assessed are sensitivity; separation of the human serum alkaline phosphatase isoenzymes of the liver, bone, and intestinal types; and differences in substrate affinity, as well as the complexity of each technique. Using the three techniques only minor differences in substrate affinity are evident. There is some nonspecific staining with the diazo-coupling technique but not with the formazan and silver staining techniques. The differences, in the mobility of the liver, bone, and intestinal isoenzymes achieved by homogeneous-pore gel electrophoresis are sufficient to allow them to be clearly distinguished. However, only very small differences in mobility are found with gradient-pore gel electrophoresis, but the sharper bands in this medium allow much smaller amounts of activity to be detected. As little as 160 microU of enzyme can be visualized by the diazo technique. Silver staining gives an approximately fourfold increase in sensitivity over the formazan technique, which in turn gives a fourfold increase over the diazo technique. An important aspect of the silver staining technique is the potential of increasing sensitivity much further by improvements in the photographic physical development stage.(ABSTRACT TRUNCATED AT 250 WORDS)

Alkaline Phosphatase↗

Fluorography of tritium-labeled proteins in silver-stained polyacrylamide gels.

Silver-staining of polyacrylamide gel electrophoresis (PAGE)-separated proteins allows sensitive detection of proteins but severely reduces the ability to detect weak beta-emitters present in the protein band. A simple procedure is described in which silver can be removed from a silver-stained PAGE gel (deargentation) using photographic fixer, and the silver-free gel can be enhanced and used for fluorography. A quantitative study of sensitivity is reported for 3H-labeled bovine serum albumin with a one-dimensional sodium dodecyl sulfate-PAGE slab gel.

Densitometry↗

Light microscopic localization of silver-enhanced liposome-entrapped colloidal gold in mouse tissues.

Silver-enhanced liposome-entrapped colloidal gold was developed for light microscopic localization of liposomes. Preparation of colloidal gold entrapped in liposomes was achieved by a modified method of Hong, et al. (1983) Biochim. Biophys. Acta 732, 320-323). In this report, a gold chloride/citrate solution of low pH (3.4) was used to inhibit the formation of gold granules during the liposome preparation. The diameter of most liposomes ranged from 80 to 100 nm. Following liposome preparation, the pH was adjusted to 6, and the temperature increased to 55 degrees C. The majority of the liposomes contained one to three gold particles. Liposomes were injected into mice via tail vein; 24 h later, tissues were collected. Sections were processed for silver enhancement of the gold particles and examined by light microscopy. Silver-enhanced gold particles were clearly observed in both liver and implanted tumor. Localization was confirmed by electron and fluorescence microscopy. Thus, we have shown that silver enhancement of colloidal gold liposomes is a direct and sensitive method for tracing the fate of liposomes in vivo, providing minimal background interference and a good definition of various cell types.

Animals↗

Mechanism of silver staining of histones: evidence for involvement of clustered lysine residues.

Pretreatment of histones with formaldehyde markedly enhances the formation of metalic silver from ammoniacal silver ion. The rate of silver reduction was determined with different histones by spectrophotometric measurement of colloidal silver stabilized in solution, and the apparent reactivity thus determined was found to be in the decreasing order of H1 greater than H2B greater than H2A greater than H3 greater than H4. Involvement of lysine residues was suggested since this order coincides with that of lysine content of these histones. However, the exceptionally high reactivity of histone H1 can be explained only when greater contribution of clustered lysine residues is assumed. Amino group modification and tryptic digestion studies of H1 corroborated this assumption.

Animals↗

Antibiotic-bonded PTFE vascular grafts: the effect of silver antibiotic on bioactivity following implantation.

Use of an infection-resistant vascular prosthesis has appeal for grafting in sites of potential contamination or to replace graft segments involved in low-grade infections caused by coagulase-negative staphylococci (CNS). In this study, antibiotic retention on polytetrafluoroethylene (PTFE) grafts prepared using three antibiotic-bonding methods was compared following implantation into the arterial circulation. Ciprofloxacin or silver-ciprofloxacin was bonded to PTFE surfaces by surfactant-mediated or direct bonding methods. Antibiotic-bonded grafts of each type (n = 8) or control grafts (n = 2) were used to replace the carotid or femoral arteries of 10 dogs. Duplicate grafts segments (0.5 x 0.5 cm) were explanted at various time intervals ranging from 1 hr to 14 days later and imprinted onto culture plates of Klebsiella pneumonia. Antibiotic retention (micrograms/ml) was determined by measuring zones of inhibition and correlated with known disc concentrations of ciprofloxacin. Antibacterial bioactivity of grafts bonded with silver-antibiotic complexes was superior (P less than 0.02) to bonding of antibiotic alone at 3 and 24 hr following implantation. Ciprofloxacin retention in excess of the minimum bactericidal concentration (0.5 microgram/ml) of Staphylococcus epidermidis was demonstrated on all silver-antibiotic-bonded grafts after 14 days of implantation. At 7 and 14 days following implantation, the bioactivity of all antibiotic-bonded grafts exceeded 0.125 micrograms/ml of ciprofloxacin, the minimum inhibitory concentration of S. epidermidis. Bonding of silver-ciprofloxacin on PTFE grafts provided an effective source of local antibiotic release at levels which may be useful for bypass grafting in contaminated wounds or for in situ replacement of grafts infected by CNS.

Animals↗

Silver accumulations in periapical granulomas: report of five cases using the scanning electron microscope, the electron microprobe, and other complementary methods.

A series of cytologic imprints obtained from periapical granulomas were studied with the conventional light microscope, the scanning electron microscope, and the electron microprobe to analyze the chemical composition of several black deposits that were randomly observed near or in the cytoplasm of macrophages and multinucleated giant cells. These granulomas had been removed from endodontically treated human teeth in which the root canals had been obturated with silver cones and Grossman's sealer 3 to 5 years previously. To investigate whether corrosion had occurred on the silver cones, the cones were also examined with the scanning electron microscope and the electron microprobe. Our observations revealed that all the examined silver cones showed different degrees of corrosion on their surfaces, whereas different concentrations of silver, sulfur, and chlorine were detected at the same sites in the cytologic imprints. However, it is impossible to determine from this study whether the presence of corrosive by-products in the periapical tissues is responsible for the development of a pathologic reaction at these sites.

Adolescent↗

Control of prolonged, benign, renal hematuria by silver nitrate instillation.

Four patients with protracted benign hematuria underwent silver nitrate instillation of the upper collecting system. The doses varied from 10 ml. of 0.25 per cent silver nitrate to 14 ml. of 1 per cent silver nitrate. In all 4 cases the hematuria resolved within four days without significant complications. At follow-up extending from three to twenty-four months there have been no recurrences of gross hematuria. Silver nitrate instillation appears to be a safe and effective method of treating benign renal hematuria, thus obviating the need for extirpative surgery.

Adult↗

Electrochemical study of endodontic silver cones used in root canal therapy.

In previous studies it was observed that endodontic silver cones placed in fine canals became dislodged as a result of corrosion. To investigate the corrosion of high purity silver, potentiostatic and potentiodynamic electrochemical techniques were used. Triangular potential sweeps made in physiological solutions and human plasma showed similar potential-current relationships. However, in human plasma, peak currents were lower and peak potentials were more anodic than those observed in the physiological solutions. The electron microprobe analysis and the EDAX of the film formed in the biological fluids revealed the presence of silver and chloride and a certain amount of carbon. The addition of small quantities of Na2S to the physiological solutions favoured metal dissolution and promoted the formation of a mixed film of AgCL and Ag2S. According to these results chloride and sulphide anions seem to be particularly aggressive towards the metal surface in implanted silver cones. Precautions to avoid direct contact of the cones with saliva and tissue fluids must be taken. Fractures and discontinuities present in the cement considerably increase the corrosion risks.

Electrochemistry↗

Transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride membranes.

We have developed a method to transfer proteins from a silver-stained polyacrylamide gel to a polyvinylidene difluoride (Immobilon-P) transfer membrane (Millipore, Bedford, MA). If the silver stained gels are rinsed in 2 x SDS Laemmli sample buffer prior to transfer, almost all proteins can be transferred comparably to non-stained controls. Some proteins stained with silver can be directly transfer, almost all proteins can be transferred comparably to non-stained controls. Some proteins stained with silver can be directly transferred to a single sheet of Immobilon-P without a prior rinse in sample buffer. Most important in the Western blot the antigenicity of the transferred protein is retained in either way. The method described is simple, inexpensive and versatile. A slight modification of the technique permits one to extract minor proteins, or detect their antigenic activities, without contamination of contiguous proteins.

Blotting, Western↗

Sensitive silver-staining detection of bacterial lipopolysaccharides in polyacrylamide gels.

For sensitive detection of bacterial lipopolysaccharides in the nanogram range, three almost identical silver-staining methods are often used, which are based on ammoniacal silver solutions and an acidic developer. We modified a method used for proteins, based on neutral silver nitrate solution and an alkaline developer, for the visualization of lipopolysaccharides in polyacrylamide gels, which yields better sensitivity and is much less prone to formation of non-specific background staining than the acidic developer-based silver stains.

Electrophoresis, Polyacrylamide Gel↗

Age-related changes in sulfide-silver staining in the rat neostriatum: a quantitative histochemical study.

The density and distribution of sulfide-silver staining in the neostriatum of 3-, 12-, and 24-month-old male Sprague-Dawley rats were analyzed using the neo-Timm sulfide-silver histochemical technique associated with microdensitometry. This technique stains zinc-containing terminals in the striatum and the density of neo-Timm staining is considered to be parallel to the density of synaptic boutons containing zinc. In the neostriatum sulfide-silver, staining was intense in the matrix, although the striosomes did not show appreciable reactivity. The density of sulfide-silver staining was significantly reduced (p less than 0.001) in the matrix of 12-month-old in comparison to 3-month-old rats. No further changes were noticeable between 24- and 12-month-old rats. In contrast, the area and the perimeter of neostriatum that were assessed by quantitative image analysis did not show age-related changes. The present results indicated that similar to the observations for a variety of neurochemical parameters of rat neostriatum such as local cerebral glucose utilization, cholinergic muscarinic receptors, and dopamine D-1 receptors, zinc-containing striatal terminal were primarily decreased between young and adult subjects but not between adult and aged animals.

Aging↗

Influence of casting procedure on silver-palladium endodontic posts.

Silver-palladium alloys have been endorsed on clinical grounds as substitutes for gold alloys in posts and cores. However, carbon absorption during heating and casting can adversely affect the mechanical properties of silver-palladium alloys. In this study the tensile characteristics of endodontic posts cast in silver-palladium alloy using either a graphite or a ceramic crucible were determined. Graphite crucibles were associated with a significantly lower (2P less than 0.00001) percentage elongation than were ceramic crucibles. The need for a carbon-free casting environment for silver-palladium alloys was confirmed. The configuration of the cylindrical, serrated, vented post used in these tests resulted in mechanical properties lower than those achieved with standard metallurgical test procedures.

Ceramics↗

Modification of silver-enhanced sodium transport across toad skin.

1. Silver stimulated short-circuit current and transepithelial potential difference. 2. Cysteine inhibited the silver-induced short-circuit current. 3. There was a dose-response inhibition of silver-induced short-circuit current by cysteine. 4. The silver-induced short-circuit current is carried by a net active sodium transfer from the outside to the inside bathing solution.

Animals↗

In vitro inactivation of silver sulphadiazine by the addition of cerium salts.

Silver sulphadiazine, with and without cerium salts, was tested in an in vitro topical antimicrobial susceptibility test assay for antimicrobial activity against microorganisms isolated from burn wounds. Compared to silver sulphadiazine alone the addition of cerium to silver sulphadiazine generally reduced the diameter of the antimicrobial inhibitory zones around susceptible organisms. In addition, complete nullification of the activity of silver sulphadiazine occurred in many instances.

Bacteria↗

Absorption, excretion and tissue distribution of silver sulphadiazine.

It was clarified that the absorption of silver sulphadiazine cream, a widely-used topical agent against Pseudomonas infection after burns, through both superficial and deep dermal burn surfaces as well as the normal skin, was negligible, but that its absorption increased when the blister was removed. After absorption, the silver sulphadiazine was dissociated into two portions. The sulphadiazine portion was excreted into the urine quickly and the silver portion, on the contrary, remained at a certain level in the body for a long time. Our results given some suggestions about the indications for silver sulphadiazine cream. The cream seems to be inadequate for treating second-degree burns but may be useful for the treatment of third- or fourth-degree burns.

Administration, Topical↗