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At least 253 records · Page 14Linked to original sources
PubMed2006

Protein stains for proteomic applications: which, when, why?

This review recollects literature data on sensitivity and dynamic range for the most commonly used colorimetric and fluorescent dyes for general protein staining, and summarizes procedures for the most common PTM-specific detection methods. It also compiles some important points to be considered in imaging and evaluation. In addition to theoretical considerations, examples are provided to illustrate differential staining of specific proteins with different detection methods. This includes a large body of original data on the comparative evaluation of several pre- and post-electrophoresis stains used in parallel on a single specimen, horse serum run in 2-DE (IPG-DALT). A number of proteins/protein spots are found to be over- or under-revealed with some of the staining procedures.

Colorimetry↗
PubMed1995

Cell death in the olfactory epithelium.

In the nervous system of vertebrates the olfactory epithelium presents unique cytological characteristics. In the olfactory mucosa, olfactory neurons die and are replaced from undifferentiated neuroblasts over the entire life span of the animal. It remains unclear whether these neurons die as a result of a direct insult from the environment or in fulfillment of a physiological program of cell death. We have studied the distribution and the characteristics of cell death in the olfactory epithelium of normal, adult rats. The olfactory epithelium contains pycnotic bodies resembling those described for thymocytes undergoing terminal apoptotic changes. These appear at all levels in the epithelium, under both light and electron microscopes and can also be demonstrated after vital staining with acridine orange. Chromatin condensation into large blocks, often located at the nuclear periphery, is a morphological hallmark of the nuclei of mature olfactory neurons, which also present an increase in electron density of the cytoplasm. After non-radioactive in situ labeling of fragmented DNA, the nuclei of olfactory neurons are positive. Under the same reaction conditions (mild protease digestion), most of the nuclei of the supporting and basal cells are negative. In vivo incorporation of 5-bromouridine, a marker of RNA synthesis, is also lower in olfactory neurons than in basal and supporting cells. These findings suggest that olfactory neurons are committed very early to physiological cell death.

Acridine Orange↗
PubMed2005

Ability of 13 chemical agents used in dental practice to induce sister-chromatid exchanges in Syrian hamster embryo cells.

To evaluate the genotoxic potential of 13 chemical agents used in dental practice, the abilities of these agents to induce sister-chromatid exchanges (SCEs) were examined using Syrian hamster embryo (SHE) cells. Statistically significant increases in the frequencies of SCEs were observed in SHE cells treated with all seven of the chemical agents used as endodontic medicaments: p-chlorophenol, m-cresol, formaldehyde, guaiacol, hydrogen peroxide, p-phenolsulfonic acid, and sodium hypochlorite (P < 0.01; Student t test). Assessment of two chemical agents that are applied to the oral mucosa as antiseptics showed that SCEs were induced by iodine (P < 0.01), but not by chlorhexidine. Of three chemical agents that are used as dyes for disclosing dental plaque, erythrosine B had no effect on SCE induction, while acid fuchsin and basic fuchsin increased the SCE frequencies in SHE cells (P < 0.01). Glutaraldehyde, which is used as a disinfectant for dental instruments and impressions, also induced SCEs (P < 0.01). Because SCE assays are used as a sensitive indicator for evaluating genetic toxicity of chemicals, the chemical agents that had a positive response in the present study are potentially genotoxic to mammalian cells.

Animals↗
PubMed1999

Presence of oxytalan fibers in human regenerated periodontal ligament.

The aim of the present study was to investigate whether oxytalan fibers are formed in the regenerated human periodontal ligament. 6 patients, each of them exhibiting an advanced intrabony defect, were treated with a bioresorbable membrane according to the GTR-principle. Following a healing period of 6 months, the teeth were extracted together with their surrounding soft and hard tissues and subsequently fixed in 10% buffered formalin. Following decalcification in EDTA, the specimens were embedded in paraffin and 8-microm histological sections were cut in the mesio-distal direction, parallel to the long axes of the teeth. The sections were stained with hematoxylin and eosin, or with the oxone-aldehyde-fuchsin-Halmi staining method and examined in the light microscope. A regenerated periodontal ligament containing newly-formed oxytalan fibers was observed in all specimens. Many of them inserted into the newly formed cementum on the root surface. It is concluded that oxytalan fibers are formed de novo in human regenerated periodontal ligament tissue.

Absorbable Implants↗
PubMed1985

Mechanism of dye response and interference in the Bradford protein assay.

Bradford Coomassie brilliant blue G-250 protein-binding dye exists in three forms: cationic, neutral, and anionic. Although the anion is not freely present at the dye reagent pH, it is this form that complexes with protein. Dye binding requires a macromolecular form with certain reactive functional groups. Interactions are chiefly with arginine rather than primary amino groups; the other basic (His, Lys) and aromatic residues (Try, Tyr, and Phe) give slight responses. The binding behavior is attributed to Van der Waals forces and hydrophobic interactions. Assay interference by bases, detergents, and other compounds are explained in terms of their effects upon the equilibria between the three dye forms.

Hydrogen-Ion Concentration↗
PubMed1985

Quantitative determination of Coomassie R bound to proteins in polyacrylamide gel. A facilitated method for multi-spot analysis of electrograms.

Dimethylsulfoxide (DMSO) was found to be an efficient solvent for extraction of Coomassie Blue R 250 (Coomassie R) from stained proteins on polyacrylamide gels. Kinetic measurements show that the extraction of the dye from a 2-D gel reached equilibrium in 48 h. Staining of E. coli ribosomal proteins by Coomassie R dissolved in trichloroacetic acid exhibited two types of dye-protein complexes, the majority of them yield a blue-purple colour, while the rest are stained with a light-blue tone and fluorescent appearance as well. The absorbance spectra of the complexes in the gel matrix differ significantly from each other. However, the DMSO-extracted Coomassie show identical absorbance profiles with lambda max at 602 nm, thus the amount of the bound dye can easily be measured spectrophotometrically.

Electrophoresis, Polyacrylamide Gel↗
PubMed1999

Carbon dye as an adjunct to isosulfan blue dye for sentinel lymph node dissection.

BACKGROUND: The success of intraoperative lymphatic mapping depends on accurate identification of the sentinel node. We hypothesized that a carbon particle suspension would allow histopathologic confirmation of the sentinel lymph node through deposition of carbon within that node. METHODS: An animal model was used to compare the lymphatic mapping accuracy of carbon dye with that of isosulfan blue dye, the standard agent for intraoperative visualization of the sentinel lymph node. Twenty-two rats underwent lymphatic mapping in each distal lower extremity with various combinations of carbon dye and isosulfan blue dye. All stained (blue or black) nodes in the inguinal drainage basin were removed for pathologic analysis, including carbon particle analysis. A meticulous search identified all nonstained (nonsentinel) nodes in the same basin. These nonsentinel nodes were examined for carbon particles by light microscopy. Dermal diffusion of mapping agents at the injection site was also recorded. Animals were then observed for 28 days to assess the toxicity of mapping agents. RESULTS: Although isosulfan blue dye and full-strength carbon dye each stained all sentinel nodes, the latter obscured histologic detail. The combination of 2.5% carbon dye, 7.5% saline solution, and 90% isosulfan blue dye also stained all sentinel nodes; carbon particles were seen on light microscopy in all 13 stained nodes and did not interfere with histologic evaluation. No unstained node contained carbon particles, although the number of nonsentinel nodes was small. Carbon dye exhibited significantly less intradermal diffusion than isosulfan blue dye, but the carbon left a permanent mark on the skin. No toxicity or side effect associated with the use of carbon dye was observed. CONCLUSION: Carbon dye allows histopathologic confirmation of sentinel lymph nodes identified by isosulfan blue dye.

Animals↗
PubMedDate not supplied

Dyes in dermatology.

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Administration, Cutaneous↗