Mucopolysaccharide staining

Recently Heremans demonstrated mucopolysaccharides in urine after zone electrophoresis by means of an Alcian blue-acid fuchsin staining technique (HI).3 Applied in our laboratory to a curtain after a run of serum, two fractions, references -)- 86.30 and - - 86.12, were demonstrated but do not seem to be regular components of serum, at least not in that high concentration (Fig. 62c). 

In addition to the methods described above a battery of other staining procedures are available. These include use of alcian blue (22) to stain glycoproteins, ethidium bromide (23) to stain DNA, and methylene blue (14) and pyronine (16) to stain RNA. A relatively new stain has been nicknamed stains-all, because of its ability to stain most macromolecules. This dye is a cationic carbocyanine and stains RNA bluish purple, DNA blue, protein red, acid mucopolysaccharides various shades of blue to purple, and phosphoproteins blue (24). It is presently the most widely used stain for RNA. 

The urine of patients with gargoylism, of both the autosomal recessive and sex-linked types, contains abnormally high amounts of acid mucopolysaccharides. Determination (T2) of the acid mucopolysaccharides in urine in relationship to preformed creatinine is useful in distinguishing certain cases of gargoylism from other conditions with which they are sometimes clinically confused (e.g., cretinism and chondro-osteo-dystrophies of the Morquio-Brailsford type). Another simple diagnostic test for gargoylism is based on the presence in the lymphocytes of abnormal inclusions of acid mucopolysaccharides which are revealed by staining (M24). 

An acid mucopolysaccharide, which was degraded by hyaluronidase, was present in abnormal amount in spaces of the trabecular meshwork in eyes containing melanomas, probably due to an alteration of the mucopolysaccharides in the vitreous (D13). In contrast to normal conjunctivitis, smears from eyes with vernal conjunctivitis are stained with... 

Numerous reports, which rely chiefly on histochemical tests, have indicated that in pathological states of skin, especially in inflammation and malignancies, there is an increase in the level of acid mucopolysaccharides. High mucopolysaccharide contents of biopsy skin sections were demonstrated in chronic lupus (MIO) and in some cases of venous edema (Zl)j in scleroderma, however, the mucopolysaccharide content of the affected skin was low (MIO). Skin lesions from patients with lupus erythematosus, dermatomyositis, lichen sclerosus et atro-phicus, and poikiloderma showed significant increases in material which was stained histochemically (S20). In two cases of urticaria pigmentosa, hyaluronic acid was found in the histologically abundant mast cells (II) injection of hyaluronidase into the involved skin areas apparently cured the patients. An increased concentration of acid mucopolysaccharides has been obseri ed in sun-damaged skin (G5, SI, S12, S18). 

Rienits later studied the zone-electrophoretic behavior of hyaluronic acid, chondroitin hydrogen sulfate, and heparin on paper strips in nonborate buffers he found that, whereas hyaluronic acid can be separated from chondroitin hydrogen sulfate and heparin, the latter two mucopolysaccharides cannot be separated. The mucopolysaccharides could be located by the use of a method similar to that of Gardell, Gordon and Aqvist, or by staining the pherogram with Toluidine Blue. The mobility of hyalu-... 

Q.29.8 What histochemical stains would be useful for identifying the following substances in a biological specimen (a) lipid containing organelles (b) amyloid fibrils (c) acidic mucopolysaccharides (d) nucleic acids (e) collagen fibers. 

When the sulphate groups are removed using the procedures described by Araki (A2), the adsorption effects as well as the electro-osmotic flow are considerably reduced. As a consequence, P-lipoproteins move at nearly the same rate as in paper-electrophoresis experiments. Moreover, sulphate-free agar is very suitable to electrophoretic analysis of mucopolysaccharides, as the background of electrophoresis diagrams stained by metachromatic techniques is practically colorless (R.E.P.A. Ballieux, personal communication). As stained electrophoresis diagrams... 

Heparinoids and mucopolysaccharides react with, and modify, many of the plasma proteins. Heparin combines with fibrinogen, globulins and albumin. As judged by electrophoresis and various types of analysis and staining, the particular plasma protein components with which heparin combines are dependent upon the concentration of protein, concentration of heparin, pH value, and salts present. This explains the somewhat contradictory statements in literature about combinations of heparin with plasma proteins. The combination may result in change of solubility of the protein and reverse protein tests . Heparin can modify the murexide reaction for calcium in serum by affecting the calcium-protein-heparin complex. Many heparinoids... 

Metachromasia, from meta, a change in the kind of, and chroma, color, refers to the qualitative change which occurs in the color of certain dyes when they interact with other substances. The term was originally applied by Paul Ehrlich to the phenomenon of color change when a dye is adsorbed onto a substrate. Thus, cartilege and other mucopolysaccharide-containing tissues are stained red by toluidine blue. Similarly, Hartley observed that when bromophenol blue solution is added to cetrimide solution the color changes from purplish blue to clear blue. 

Bronchiolitis obliterans (BO) with intraluminal polyps is characterized by the presence of buds or polyps of granulation tissue projecting or completely filling the lumens of membranous and/or respiratory bronchioles. These polyps can have a myxoid or pale staining matrix (rich in acid mucopolysaccharides) in which elongated myofibroblasts and inflammatory cells are embedded or they can be richer in collagen fibers. 

The Alcian Blue test. Urine is spotted on to filter paper and immersed in a solution of Alcian Blue. If mucopolysaccharides are present, the urine spot stains blue. 

Mast cells are connective-tissue cells widely distributed throughout the body, particularly in the lungs and liver, although there are places where they are not found such as the wall of the stomach. Mast cells are detected by metachronuitic staining due to their high concentration of an important mucopolysaccharide, heparin virtually all endogenous heparin is located in the mast-cell system. In... 

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