SEARCH · Search PubMed
Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.
Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.
A method for typing Aeromonas species by silver staining of total soluble proteins separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis is described. There was good agreement with the results obtained by autoradiography of whole-cell proteins for isolates examined by both methods.
The numbers of silver-stained nucleolar proteins (AgNORs) were counted in hyperplastic and neoplastic lesions of the endometrium and compared with those of normal proliferative and secretory phase endometrium. In glandular cells in the normal menstrual cycle, the mean number of AgNORs in proliferative phase endometrium (3.8) was significantly higher than that in secretory phase endometrium (2.7, P less than 0.05). The mean number of AgNORs in well-differentiated endometrioid type adenocarcinoma (5.5) was significantly higher than that in both complex hyperplasia without cytological atypia (3.6, P less than 0.01), and simple hyperplasia (3.3, P less than 0.01). Mean AgNOR counts in complex hyperplasia with cytological atypia were greater than those in both complex hyperplasia without cytological atypia (P less than 0.05) and simple hyperplasia (P less than 0.05). Thus, complex hyperplasia with cytological atypia appears to be a direct precursor of well-differentiated endometrioid type adenocarcinoma. These findings suggest that the mean numbers of AgNORs are increased in neoplastic changes in the endometrium, and the one-step colloid method for AgNORs may therefore be a simple and useful technique to examine proliferative activity in neoplastic and pre-neoplastic endometrial cells.
The karyotype of the aphid Aphis sambuci has been studied by means of C-banding, fluorochrome staining and AgNO3 staining. Heterochromatic areas are confined exclusively to the X chromosomes. Interestingly, silver staining produces a clear-cut longitudinal differentiation of all chromosomes, which allows reliable identification of homologues.
BACKGROUND: The diagnosis of pulmonary involvement in hematologic malignancies (PHM) has traditionally necessitated large specimens (usually invasive or surgically obtained biopsies). The development of reliable immunocytologic identification techniques allows diagnosis on noninvasively obtained cellular material, including bronchoalveolar lavage (BAL) fluid. Since 1985, 12 such cases have been reported. CASES: Three cases of B-cell-related PHM were diagnosed by means of BAL fluid cellular analysis using the immunogold-silver staining technique. CONCLUSION: This is the first report showing immunogold-silver staining to be a reliable immunocytochemical identification technique for monoclonal B-cell populations recovered by BAL.
The silver-staining technique was used to study even cases of structural anomalies involving acrocentric chromosomes, excluding Robertsonian translocations. This technique allows identification of breakpoints and furnishes some information about interphase cell dynamics not seen on conventional chromosome preparations.
Different methods for the isolation of soluble proteins were applied to cell cultures of three apple cultivars (Malus sylvestris Mill.), best results being obtained with a rapid technique based on freezing and thawing. Ultrathin-layer isoelectric focusing followed by an improved silver staining method has shown that proteins from apple callus cultures consist of some 60 to 80 zones, with isoelectric points mainly between pH 4 to 7. Depending on protein content, adequate silver staining is achieved with 50 to 500 cells. Protein patterns of callus cultures allowed clear discrimination of cultivars. Protein and peroxidase isozyme patterns in cell saps of suspension cultures show striking differences during the growth cycle, whereas the protein patterns from the nutrient media were constant over the entire cultivation period and closely resembled the patterns of stationary phase and callus cells.
A ten-minute methenamine silver staining procedure for the rapid demonstration of Pneumocystis carinii and fungi in tissue sections has been developed. The procedure is optimal for both urgent and routine histologic diagnosis of diffuse, progressive, and life-threatening pulmonary infiltrates, especially in the compromised host, and of fungal lesions in general. Moreover, it is simple and brief, is applicable to both paraffined and frozen sections, and can easily be used by the pathology resident or pathologist if the need arises. It consistently yields results that are far superior to those obtained by Grocott's original methenamine silver method, and as good as or superior to those of modified silver methods and such nonsilver procedures as Giemsa's, Gram-Weigert, and toluidine blue stains.
A combination of the silver-staining method of the nucleolus organizer regions (NORs) with a Giemsa-banding method is described. This double staining allows a rapid identification of the NOR-bearing chromosomes.
A simple and sensitive technique to detect point mutation of the p53 gene using silver staining method for single-strand conformation polymorphism analysis of polymerase chain reaction (PCR-SSCP) is reported. In this study, the aberrations of the p53 gene (exon 6-8) in clinical specimens of primary cervical carcinoma were examined. Of 27 tumor tissues 2 samples showed mutations of p53 exon 7. Absence of abnormal bands in the p53 exon 6-8 in all ten normal cervical tissues. It appears that inactivation of p53 gene by point mutation is infrequent in clinical samples of human cervical carcinomas.
Scattering of the silver-stained proteins of nucleolar organizer regions (Ag-NOR proteins) was produced by actinomycin D in Ishikawa cells. Scattering of Ag-NOR proteins was found only in cells treated with actinomycin D and various other agents had no effect. Scattering was dose-dependent up to 10(-2) micrograms/ml of actinomycin D, but it was not found at higher concentrations that caused marked inhibition of total DNA and RNA synthesis. Actinomycin D (10(-2) micrograms/ml) caused the following changes: (i) nucleolar segregation and (ii) emergence of dense fibrillar bodies in the nucleoplasm. Ag-NOR proteins were observed on the fibrillar centers and surrounding fibrillar components in control nucleoli, on the fibrillar and amorphous zones in segregated nucleoli, and on the dense fibrillar bodies emerging in the nucleoplasm. The scattering of Ag-NOR proteins was due to the argyrophilic nature of the dense fibrillar bodies. Actinomycin D (10(-1) micrograms/ml) also caused similar morphological alterations in the nucleolus and nucleoplasm, but Ag-NOR proteins were observed only on nucleolar remnants.
We investigated the effects of sex steroids on silver-stained nucleolar organizer regions (Ag-NORs) and DNA/RNA kinetics in Ishikawa cells. Norethindrone and its isomer norethynodrel exclusively caused Ag-NORs to scatter in the nuclear matrix, from the nucleolus. No such effect occurred with the other sex steroids tested, including progestational, androgenic, and estrogenic compounds. Nuclear argyrophilic substances induced by norethindrone, as well as nucleolar ones, were neither DNA nor RNA but protein. Electron microscopy showed that norethindrone caused nucleolar segregation, in which the fibrillar components disappeared, and it produced islets, consisting of dense fibrillar materials, in the nucleoplasm. Ag-NORs observed on the fibrillar components in control nucleoli were translocated onto the dense fibrillar materials in the nucleoplasm. Although scattering was preferentially found in the cells synthesizing DNA, the scintillation assay of DNA/RNA kinetics suggested that scattering was related to the inhibition of RNA synthesis. These results imply that norethindrone preferentially interacts with intranucleolar DNA when its duplication is occurring and then interferes with rRNA synthesis. Scattering of Ag-NORs might not be caused by the hormonal activity of these agents but by a pharmacological effect derived from their molecular structures.
A system for the simultaneous silver staining of numerous two-dimensional gels bound to polyester sheets is proposed. Under controlled conditions, a very good reproducibility of staining can be obtained, even between different experiments. Procedures to avoid artifacts due to polyester sheets (background, silver mirror, and vertical streaking) are described.
Two hundred and eighty two specimens from 220 patients positive for HIV with respiratory tract symptoms, or febrile illness, or both, were examined for the presence of Pneumocystis carinii. Specimens were either induced sputum samples or bronchoalveolar lavage fluids. To establish the optimal method for laboratory diagnosis a comparison was made of detection of the organism by use of monoclonal antibody and immunofluorescence with conventional silver staining methods. Three commercially available reagents for immunofluorescence were also compared. Immunofluorescence was significantly more sensitive than the silver stain and the best results for immunofluorescence were obtained using. Northumbria Biologicals Ltd reagents.
A procedure for preparing duplicate images of polyacrylamide gels containing silver-stained proteins using X-ray direct duplicating film is described. Resulting images are shown to be qualitatively and quantitatively faithful representations of the original gel.
In this paper we reported the preliminary application of immunogold-silver staining (IGSS) technique in immunopathological diagnosis of thromboangiitis obliterans (TAO). We used IGSS technique to detect the immune complexes deposit in the vascular walls (18 cases) and the anti-vascular autoantibody in serum (28 cases) of TAO patients. The immune complexes deposited in tissue were found in 100%, and the anti-vascular autoantibody in 86% of TAO patients. The results of IGSS technique were compared with that of immunofluorescent staining, immunoenzyme staining and avidin-biotin-peroxidase complex staining techniques. It was demonstrated that both the sensitivity and specificity of IGSS technique were significantly higher than those of other techniques.
The potential of immunogold-silver staining for study of leukocyte subpopulations, as defined by monoclonal antibodies in cell suspensions, was examined. The cells were labeled in suspension as described for immunogold staining. Cytocentrifuge preparations of the suspensions were then immersed in a physical developer. By light microscopy, cells reacting with the monoclonal antibodies showed dark granules on their surface membrane. The morphology of the cells, as revealed by a panoptic counterstain, was comparable with that seen in routine cell smears for differential counts. The numbers of T-cells, T-helper/inducer cells, and T-cytotoxic/suppressor cells counted by this method in normal peripheral blood were nearly identical to those identified by immunogold staining and immunofluorescence microscopy in the same cell suspensions. The good morphological delineation also made possible rapid and accurate identification of particular leukocyte subsets in complex cell suspensions. Atypical lymphocytes from patients with infectious mononucleosis displayed the surface phenotype of activated T-cytotoxic/suppressor cells. Different maturation stages of neoplastic cells in patients with acute myeloid leukemia showed differences in surface antigen expression. Immunological detection of cell surface antigens could be combined with cytochemical staining of intracellular enzymatic activities. Finally, the labeling could be performed on cells prefixed on glass slides.
Immuno-gold and silver staining (IGSS) is an advanced labelling method followed by Immuno-fluorescence(IF),Immunoenzyme and Radio-immunity.In this study,the IGSS method for detection of deposits of Immunogloblins (IgG,IgA,IgM) at the BMZ had been performed on biopsy specimens from oral lesion of patients with OLP and DLE.The results indicated that the sensitivity and specificity of IGSS were super than that of DIF.It suggested the IGSS be an available method to detect oral mucosal diseases widely.
The quantitative attributes of human leukocyte proteins detected by silver staining two-dimensional electrophoresis (2-DE) gels were studied by using computer-assisted data analysis. Experiments included (a) analysis of replicate patterns of the same sample, (b) analysis of different dilutions of the same sample, and (c) analysis of samples from different individuals. Over 200 proteins were observed to have coefficients of variation (CV) less than or equal to 15% when data from replicate patterns were analyzed. In contrast, 8 proteins had CV values of less than or equal to 15% when data from different samples were analyzed. The dilution experiment showed that a majority of the proteins detected with some consistency (i.e., observed in at least 80% of the patterns) have a linear relationship between the amount of protein loaded onto a 2-DE gel and the spot volume in the final 2-DE pattern. The slope of the curves and the deviation from linearity were found to be quite protein-specific. These results indicate that optimization of sample purity and minimization of staining protocol variables are required to limit the background quantitative variability between and within 2-DE runs to a level that will allow detection of quantitative changes indicative of biological responses.