alcian-blue and Hyperplasia

Determination of hyperplastic and hypertrophic changes of mucus-secreting cells in animal airways has been performed in the past by using histologic, immunologic, and/or molecular biologic approaches. Histologic techniques are tedious and time-consuming. The other approaches require specific antibodies and cDNA probes that have proved difficult to develop. Described here is a method for the rapid estimation of hyperplastic and hypertrophic changes of secretory epithelial cells in rat airways. The assay specifically measures acidic and neutral mucoproteins in a linear fashion from 0.5 microgram to at least 10 micrograms. Male Sprague-Dawley rats were exposed to metabisulfite mist (10% wt/vol) for 5 days/wk for 3 wk. The lungs were removed and homogenized in a phosphate-buffered solution containing reducing agents and protease inhibitors. The particulate matter was removed by centrifugation, and the soluble extract was applied to a column packed with Sepharose CL-6B. The material eluting in the void volume was applied to a PVDF membrane and stained for either acidic or neutral mucosubstances using Alcian blue or periodic acid-Schiff (PAS) staining, and the absorbance was read using a 96-well plate reader. Lungs from sodium metabisulfite-exposed animals showed a 7-fold and 3.5-fold increase in PAS-positive and Alcian blue-positive material, respectively. The increase in both PAS and Alcian blue staining was hyaluronidase and chondroitinase insensitive. The observed changes are consistent with morphometric measurements of mucus-containing cells in histologic sections of the tissues. This assay may be useful in determining which neurohumoral mediators might be involved in mucus cell hypertrophy and hyperplasia in animal models of chronic obstructive pulmonary disease.

Topics: Administration, Inhalation; Alcian Blue; Animals; Chromatography, Agarose; Epithelium; Histocytochemistry; Hyperplasia; Hypertrophy; Lung; Male; Mucins; Periodic Acid-Schiff Reaction; Proteoglycans; Rats; Rats, Sprague-Dawley; Staining and Labeling; Sulfites; Trachea

Clear cell metaplasia of the human breast is known to be a benign metaplastic change which has no pre-malignant connotation. Despite its proposed relationship to focal lactational change and to lactating breast, morphological and immunocytochemical features failed to demonstrate a clear relationship between these. Mucin secretion showed a characteristic pattern of granularity, and endocrine differentiation was not present. The mucin and immunocytochemical features suggest a relationship with eccrine sweat glands and a better name would perhaps be 'eccrine metaplasia' to underline the special relationship breast metaplasias have to sweat gland epithelium.

Topics: Alcian Blue; Apolipoproteins; Apolipoproteins D; Breast; Carrier Proteins; Eccrine Glands; Female; Glycoproteins; Humans; Hyperplasia; Immunohistochemistry; Keratins; Lactalbumin; Lactation; Membrane Transport Proteins; Metaplasia; Neoplasm Proteins; Pregnancy; S100 Proteins; Sweat Glands

The glycocalyx of absorptive cells in large intestinal hyperplastic polyp was characterized histochemically at the electron microscope level by the use of the Alcian Blue pH 2.5 and high iron diamine techniques with the aim of comparing their ability in preserving the fine reticular network of the structure. Both the reagents stained glycocalyx, indicating the presence of sulphated acidic glycoconjugates. However, they showed different degrees of condensation of the reactive sites. Alcian Blue preserved its filamentous appearance better.

Topics: Alcian Blue; Colonic Polyps; Diamines; Histocytochemistry; Humans; Hyperplasia; Iron; Microscopy, Electron; Staining and Labeling

Biopsies of duodenal and ileal mucosa from patients with familial polyposis coli were studied. Areas of atypia were identified in the duodenum of six patients and in the ileum of three patients. Grade I atypia was characterized by crowding and elongation of cells and nuclei, a slight reduction in the number of goblet cells and the presence of a brush border; grade II atypia was further characterized by pseudo- or pluristratification of cells, a marked reduction in the number of goblet cells and the absence of a brush border. In areas of atypia, columnar cells often contained PAS-positive apical granules, which were diastase-resistant and unstained by alcian blue at any pH; the brush border, even where recognizable in haematoxylin-eosin and PAS-stained sections, was unreactive histochemically for alkaline phosphatase. Goblet cells were few in areas of atypia, but those present were regularly stained by PAS and alcian blue pH 2.6. Apical granules, similar in their histochemical characteristics to those observed in columnar cells in areas of atypia, were also found in otherwise normal mucosal areas, even in some patients with no overt areas of atypia in the biopsies studied. These granules have been interpreted as an abnormality, possibly preceding the onset of atypia. Hyperplasia of goblet cells, secreting mucins with the same staining pattern as in normal intestine, was found in some patients, either adjacent to areas of atypia or independent of them. Intervening columnar cells had a normal morphology, alkaline phosphatase-reactive brush borders and no sign of mucus secretion. This goblet cell hyperplasia has been interpreted as a reactive, nonspecific alteration of the mucosa.

Topics: Adenomatous Polyposis Coli; Adolescent; Adult; Aged; Alcian Blue; Biopsy; Duodenum; Histocytochemistry; Humans; Hyperplasia; Ileum; Intestinal Mucosa; Middle Aged; Periodic Acid-Schiff Reaction