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PubMed2006

Fabrication of protein-conjugated silver sulfide nanorods in the bovine serum albumin solution.

Highly ordered silver sulfide nanorods conjugated with the Bovine Serum Albumin (BSA) protein have been successfully achieved at ambient temperature. Such a process is very simple and controllable, directly using silver nitrate and thioacetamide (TAA) as the reactants in the aqueous solution of BSA. The products have been characterized by XRD, HRTEM-SAED, SEM-EDS, TG-DTA, FT-IR, and CD spectroscopy. The results of the research show that the as-prepared Ag2S nanorods are monodispersed with sizes about 40 nm in diameter and 220 nm in length, and exhibit a high degree of crystallinity and good photoluminescence. Furthermore, an interesting mechanism is discussed for the formation of the Ag2S nanorods.

Microscopy, Electron, Scanning↗
PubMed1995

Intra-species characterization of clinical isolates and biotechnologically used strains of Lactobacillus rhamnosus by analysis of the total soluble cytoplasmatic proteins with silver staining.

Lactobacillus rhamnosus is often used in milk products because of its technological properties, however it is supposed to be related to human infections as well. Therefore, 23 L. rhamnosus strains, including the type strain, were investigated. Nine strains resulted from biotechnological sources, 14 isolates had a clinical background, 13 came from a swedish culture collection. Biochemical and physiological properties were proved by classical tests. Analysis of the total soluble cytoplasmatic protein patterns was performed with diamine silver staining, a technique not previously applied to lactobacilli. Classical tests were able to confirm all strains as L. rhamnosus. Analysis of protein patterns allowed to differentiate between three clusters. Cluster I contained the type strain and biotechnologically used strains and one clinical strain isolated in Berlin. The second consisted of both, technological and clinical strains. The third cluster contained clinical isolates alone. SDS-PAGE of proteins together with diamine silver staining seems to be helpful to detect intra-species differences. It was stated, that technologically used strains of L. rhamnosus could clearly be differentiated from each other and also from clinical isolates.

Bacterial Proteins↗
PubMed1986

[Advantages and disadvantages of silver staining of proteins in polyacrylamide gel].

Sensitivity of protein staining with Serva blue G-250 (Coomassie brilliant blue G-250 analogue) in polyacrylamide gel was determined. It has been shown that protein staining with 0.1% Serva blue G-250 results in the recovery of 80 to 35 ng of single protein, that is almost 10 times higher than reported previously for Coomassie brill. blue G-250 (or R-250) staining. The comparison of the sensitivity of Serva blue G-250 protein staining in PAAG and AgNO3 has shown that AgNO3 staining was approximately 18-30 (but not 100 times, as it had been thought before) times more effective for the majority of proteins under study. Silver staining of some proteins, for instance ribonuclease and a number of retrovirus-specific structural proteins, was of lower efficacy. Thus, to obtain reliable results protein electrophoresis in PAAG should be followed by both staining procedures.

Electrophoresis, Polyacrylamide Gel↗
PubMedDate not supplied

Electron microscopic localization of silver binding proteins in the cytoplasmic granules of guinea pig eosinophil.

A new electron optic configuration is described in the specific granules of eosinophil leukocytes. Application of the ammonium silver nitrate impregnation after treatment of intact cells with formaldehyde-OsO4 solution reveals regular deposits of 50 A-electron-opaque particles which are localized in the axial region of the central core. They contain acid-extractable proteins that seem to include the eosinophil basic proteins which participate to the local immune response of the host.

Animals↗
PubMedDate not supplied

[Demonstration of viral proteins by silver staining].

Investigations were conducted to establish the optimal conditions for silver staining of polypeptides separated by polyacryle-amide gel electrophoresis. The thickness of the gel layer was of 1.5 mm. The experiments showed that the modified Hankeshover and Dernick's technique (4) (replacement of the fixative for the electrophoretically separated polypeptides and extension of the bleaching by Farmer's reagent time) had the best sensitivity and reproducibility. This technique can be performed also on polyacryle-amide gel preparations already stained by the Coomassie Brilliant Blue method. To check the efficiency of this silver staining technique, the authors used it to detect the Sendai virus and B hepatitis virus surface antigen polypeptides.

Electrophoresis, Polyacrylamide Gel↗
PubMed1994

Quantification of silver-stained proteins resolved by two-dimensional electrophoresis: genetic variability as related to abundance and solubility in two maize lines.

Relative abundance and solubility of proteins from etiolated coleoptiles of maize were investigated using two-dimensional electrophoresis (2-D PAGE). Automatic quantification of silver-stained polypeptides on replicate 2-D gels made it possible to test the linearity of the relationship between spot integrated optical density and protein amount in the range from 15 micrograms to 135 micrograms per gel, in two inbred lines. A linear response was found for more than 60% of the spots in each genotype. When a linear response was found in both lines for a given spot, the slope values were similar in 94% of the cases, indicating the reliability of silver staining for polypeptide quantification. The parameters of the curves allowed the definition of protein classes of different abundances that could be compared for genetic variability between the two lines. From a comparison between standard 2-D patterns (trichloroacetic acid-acetone extracted proteins) and patterns obtained from Tris buffer extracted proteins, it appeared that 92% of the proteins visualized in the standard gels were soluble. No difference in genetic variability, either qualitative or quantitative, was evidenced between the various classes of abundance, or between soluble and insoluble proteins.

Analysis of Variance↗
PubMed1993

Muramidase-mediated damage to Candida yeast cells. Histochemical and immunochemical characterization of accumulating wall-like material.

Hen egg-white lysozyme is known to be fungicidal to blastoconidia of Candida albicans under defined in vitro conditions. This lethal action leads to changes in the layering of cell wall and to plasmolysis, caused by unremitting accumulation of wall-like material between the yeast cell wall and cytoplasmic membrane. Here, several methods were applied on ultrathin sections to define the nature of wall-like material: histochemical staining with periodic acid-thiocarbohydrazide-silver proteinate, periodic acid-alkaline bismuth, and phosphotungstic acid at low pH; the localization of the carbohydrate residues with lectin-gold complex; immunocytochemical staining with monospecific antibodies, factor 1 and 6, which recognized major cell wall antigens. The wall-like material was almost uniformly highlighted with periodic acid-thiocarbohydrazide-silver proteinate, factor 1 antibody, concanavalin A-gold and wheat germ agglutinin-ovomucoid-gold, indicating the presence of mannoproteins and chitin. The serotype A-specific epitope recognized by factor 6 antibody was not detected in the wall-like material, although it was demonstrated in the outer cell wall layers after 2 h of exposure to lysozyme.

Antibodies, Monoclonal↗
PubMed1991

Prophylaxis of endophthalmitis with topical povidone-iodine.

The authors conducted an open-label nonrandomized parallel trial to examine whether the preoperative application of povidone-iodine to the ocular surface reduces the incidence of endophthalmitis after intraocular surgery. During an 11-month period, topical 5% povidone-iodine was used to prepare the conjunctiva in 1 set of 5 operating rooms, while silver protein solution was used in another set of 5 rooms. In all cases, surgeons continued to use their customary prophylactic antibiotics. A significantly lower incidence of culture-positive endophthalmitis (P less than 0.03) was observed in the operating rooms using povidone-iodine (2 of 3489 or 0.06%) compared with those using silver protein solution (11 of 4594 or 0.24%). Use of topical povidone-iodine in over 3000 cases was not associated with any adverse reactions. In a majority of the observed cases of endophthalmitis, some form of intraoperative communication with the vitreous cavity existed.

Administration, Topical↗
PubMed1982

Extracellular and intracellular digestion of complex carbohydrates by osteoclasts.

The role of osteoclasts or chondroclasts in degradation and synthesis of complex carbohydrates was investigated using the high iron diamine-thiocarbohydrazide-silver proteinate method (HID-TCH-SP) for sulfated glycoconjugates and the periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method for vicinal glycol-containing glycoconjugates. HID-TCH-SP stained the calcified cartilage matrix, the osteoclast ruffled border, vacuoles and heterophagosomes but not the Golgi apparatus and primary lysosomes. The size and number of HID-TCH-SP stain deposits progressively decreased from the calcified cartilage matrix to the ruffled border (p less than 0.001). Enzyme digestion with testicular hyaluronidase removed most HID-TCH-SP stain deposits averaging 13 nm. in diameter in the extracellular matrix, cytoplasmic vacuoles, and heterophagosomes of osteoclasts. Only sparse stain deposits averaging 8 nm. in diameter and presumed to be keratan sulfate remained in these sites after enzyme digestion. Osteoclast heterophagosomes contained the highest concentration of hyaluronidase-resistant material suggesting delayed degradation of keratan sulfate at this site. PA-TCH-SP strongly stained collagen fibrils in the calcified cartilage matrix. Reactive collagen fibrils were also observed in extracellular channels but only rarely were identifiable collagen fibrils observed in cytoplasmic vacuoles. A progressive decrease in the diameter of PA-TCH-SP reactive collagen fibrils was observed between the calcified cartilage matrix and the ruffled border region (p less than 0.001). PA-TCH-SP stained cisternae of rough endoplasmic reticulum, Golgi saccules, and primary lysosomes consistent with the synthesis and packaging of glycoprotein enzymes at these sites. These results indicate that the dissolution of sulfated glycoconjugates requires osteoclastic engulfment of degraded material and subsequent intracellular digestion, whereas the dissolution of collagen fibrils appears to be completed extracellularly.

Animals↗
PubMed1992

Interaction between cortisol and cortisol-binding protein in silver foxes (Vulpes fulvus).

1. Selection of silver foxes for domestic behaviour resulted in the parallel lowering of both cortisol and cortisol-binding protein (CBP) levels in the blood plasma. 2. During seasonal cycles (summer-winter) and after stress an increase in cortisol levels is followed by a decrease in CBP activity. 3. It is concluded that there are two types of interaction between cortisol and CBP in silver foxes: parallel changes in the process of domestication and opposite changes under the influence of environmental factors.

Animals↗
PubMed1982

Substructure of granules from serous cells of porcine tracheal submucosal glands.

The ultrastructure of serous cells from porcine tracheal submucosal glands was studied by conventional transmission electron microscopy (TEM), and by cytochemical methods to stain for complex carbohydrates. In tissue fixed and processed for TEM, and stained with uranyl acetate and lead citrate, the condensing granules of serous cells occasionally possessed a hexagonal and sometimes a lamellar substructure. Tissue fixed in paraformaldehyde-glutaraldehyde and stained with periodic acid-thiocarbohydrazide-silver proteinate (PTS) or with phosphotungstic acid (PTA) showed secretory granules stained for complex carbohydrates and revealed a substructure similar to that noted in the condensing granules. The dark staining substructure revealed by either the PTS or the PTA technique appeared to correspond to electron-lucent areas observed in the condensing granules by conventional TEM. The PTS staining probably demonstrated the presence of neutral glycoprotein, since the serous-cell granules did not react with a dialyzed iron stain for acidic glycoproteins. Treatment of periodic acid oxidized thin sections with pronase or pepsin prior to thiocarbohydrazide and silver proteinate treatment decreased the intensity of the PTS staining, but did not digest away any components of the granules. The substructure revealed by the carbohydrate stains may be a reflection of the mechanism of packaging or the macromolecular structure of the glycoproteins in the serous-cell granules.

Animals↗
PubMed1981

Ultrastructural localization of complex carbohydrates in odontoblasts, predentin, and dentin.

Glycosaminoglycans (GAGs) and glycoproteins (GPs) are essential components for dentinogenesis. We have examined rat odontoblasts, predentin, and dentin decalcified with EDTA and stained with: 1) Spicer's hig-iron diamine-thiocarbohydrazide-silver proteinate (HID-TCH-SP) method for sulfated glycoconjugates, and 2) Thiéry's periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method for vicinal glycol-containing glycoconjugates. HIS-TCH-SP stained distended portions of Golgi saccules and secretory granules. The predentin contained three times the number of HID-TCH-SP stain precipitates when compared to the mineralization front of the dentin matrix. PA-TCH-SP weakly stained membranes of Golgi saccules and cisternae of rough endoplasmic reticulum (RER), whereas stronger staining was observed in secretory granules, lysosomes, and multivesicular bodies (MVBs). Collagen fibrils in predentin demonstrated moderate PA-TCH-SP staining. In contrast, strong PA-TCH-SP staining was observed on and between collagen fibrils in the mineralization front of the dentin matrix. TCH-SP controls of unosmicated specimens lacked significant staining, however, osmicated control specimens did contain some TCH-SP stain deposits in the mineralization front. These results indicate that sulfated and vicinal glycol-containing glycoconjugates are packaged in the same type of secretory granule and released into the extracellular matrix; subsequently vicinal glycol-containing glycoconjugates concentrate in the calcification front, whereas sulfated glycoconjugates accumulate in the predentin and are either removed or masked to staining in the dentin.

Animals↗