Neutral Mucosubstances Research Articles

The use of double staining techniques for investigating bacterial attachment to mucopolysaccharide-coated epithelial cells.

Double staining techniques were devised to study Escherichia coli attachment to mucopolysaccharide-coated urinary tract epithelial cells. In addition, vital stains were used to distinguish between viable and nonviable epithelial cells. Alcian blue was used to confirm the presence of glycosaminoglycans and periodic acid Schiff was used to distinguish a further group of polysaccharides, proteoglycans, neutral mucosubstances and glycolipids. The staining methods enabled investigations to be carried out concerning the possible importance of mucopolysaccharides in the attachment of bacteria to mucosal surfaces. Staining techniques were also used to investigate whether the presence of a mucopolysaccharide coat is related to epithelial cell viability. The combinations of vital and mucopolysaccharide stains were found to give reproducible results when cell preparations were evaluated by three individuals. Both in vivo and in vitro certain populations of epithelial cells have been found with a large number of bacteria preferentially attached. The double staining techniques described here may help to reveal the nature of these target cells.

Gastric mucosubstances and benign gastric ulceration.

Variations in the composition of mucosubstances have been demonstrated in the gastric contents of patients with chronic gastritis, pernicious anaemia and gastric carcinoma. There have however been few histochemical studies of the epithelial mucins observed in the benign gastric ulcers1. Neutral mucosubstances have been identified by their reaction with periodic acid-Schiff’s reagent to form magenta chromophores2. Haynes and Hartridge3 and Steedman4 have shown acidic mucosubstances to interact with Alcian Blue at different levels of pH to form blue chromophores. The method employed in this study is based on electronic densitometric measurements of chromophores. The energy absorbed by each chromophore is dependent upon two factors, the spectral characteristics of the chromophores and the concentration of the stained material.KeywordsGastric UlcerAlcian BluePernicious AnaemiaEpithelial MucinAntral BiopsyThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Observations on the histology and cytochemistry of the digestive gland inPila virens (Lamarck) (Mollusca: Gastropoda)

The histology and histochemistry of the digestive gland and the investing epidermis ofPila virens are described. The digestive gland tubule epithelium is made up of the digestive cells and calcium-cum-excretory cells. The digestive cells are columnar, vacuolated cells and carry out the function of secretion and absorption. The calcium-cum-excretory cells are pyramidal non-vacuolated cells that harbour prominent excretory pellets. These cells serve as reservoirs of calcium and also carry out the function of excretion. The investing epidermis of the gland is composed of low cuboidal epidermal cells interspersed with mucous cells that seem to secrete both neutral and acid mucosubstances. Overlying this is a melanin layer.

Mucosubstances in the colonic goblet cells of the ferret

Mucosubstances in the colonic goblet cells of the ferret (Putorius [Mustela] furo) were studied histochemically. They contained predominantly sulphated mucosubstances in the proximal colon and only sulphated mucosubstances in the distal colon. The sulphated mucosubstances were of weakly acidic nature. Some deep cells and a few superficial cells in the proximal colon contained sialomucins which were sialidase-sensitive. Neutral mucosubstances were not found in the colon.

Histochemical studies of mucosubstances in the epidermis of a nonscaly teleost Mystus (Mystus) vittatus Bl. (Pisces-Bagridae).

The epidermis of Mystus (Mystus) vittatus contains two well differentiated mucous cells which secrete different mucosubstances. The goblet cells contain periodate reactive neutral mucosubstances, glycogen, testicular hyaluronidase resistant sulphated mucosubstances, and sialic acid rich glycoproteins. The clavate cells contain small amounts of neutral and sulphated mucosubstances and no glycoproteins. The difference in the histochemical nature of the two types of mucous cells is discussed in relation to their physiological activities.

MUCOSUBSTANCES OF THE RAT GASTRIC MUCOSA STUDIED BY MEANS OF SEVERAL HISTOCHEMICAL METHODS

Distribution of mucosubstances in the rat gastric mucosa was studied by radioautography using 35S-sodium sulfate, several light microscopic histochemical procedures, the periodic acid thiocarbohydrazide silver proteinate (PA-TCH-SP) method and postembedding staining with Iectin (wheat germ agglutinin) ferritin conjugates. Histochemical staining revealed that the superficial covering epithelium contained a large amount of neutral mucosubstances, whereas the foveolar and isthmus mucous cells produced both sialomucins and sulfomucins. Active incorporation of 35S was observed in the fundic foveolus and isthmus as well as the pyloric gland. The mucous neck cells contained sialomucin. The PA-TCH-SP method stained the covering epithelium, the epithelium of the foveolar and isthmus mucous cells and the mucous neck cells. All cell organellae which took part in the formation of granule substances in the gastric mucosa were stained by this method. The cytoplasm of the mucous neck cells was extensively occupied with round PA-TCH-SP positive granules. As the epithelium of the pits matured, the granules increased in size. The apical portion of the covering epithelium was exclusively occupied with large round to oval granules. Many of them showed a bipartite structure after PA-TCH-SP staining, reaction products were observed only in the peripheral region of these granules. Conversely, the development of other cell organellae became less prominent as these epithelia matured. Postembedding staining with lectin (wheat germ agglutinin)-ferritin conjugates stained secretory granules of all these mucous cells with different staining patterns. In the covering epithelium, the conjugate stained only the peripheral region of the granules.

The fine structure of the stomach mucosa of the llama (Llama guanacoe)

The epithelium of the fundic region mucosa of the hind stomach in the Llama guanacoe has been studied using morphological and histochemical methods. Morphology suggests that solute and water absorption may occur in the epithelium of the surface and of the foveolae, although this absorption can not be estimated because of the extensive secretion of the gastric glands. The same cells of the surface and foveolar epithelium show numerous secretory granules. The glands reveal neck cells, chief cells, a large number of oxyntic cells, four types of endocrine cells (A-like, ECL, D and EC), brush cells and wandering cells. PAS and Alcian blue reactions for light microscopy suggest a secretion of neutral and acidic mucosubstances in the surface and foveolar epithelium, of neutral mucosubstances only in the neck cells. Periodic acid-thiocarbohydrazide silver proteinate (PA-TCH-SP) reaction for electron microscopy confirms the presence of neutral mucosubstances within the secretory granules of the surface, foveolar and neck epithelial cells. In all these cells, the reaction product is also evident within sacculi and vesicles of the maturing surface of the Golgi apparatus. A positive PA-TCH-SP reaction also occurs on the membrane (and not on the contents) of the Golgi apparatus (maturing surface) and of the secretory granules of the chief cells as well as on the membrane of the Golgi apparatus and of apical vesicles and tubules of the oxyntic cells. In addition, silver granules slightly enhance the electron desity of the contents of the secretory granules in the endocrine cells. Morphological and histochemical findings are discussed and compared with results described by others for monogastric mammals.

Histochemistry of mucosubstances in the lingual glands of the mongoose.

The histochemistry of mucosubstances in the lingual glands of the mongoose was studied. Both serous and mucous or mixed glands were present in the posterior part of the tongue. In serous glands, acinar cells and occasional duct cells contained granules which gave reactions for neutral mucosubstances only. In mucous glands, tubular cells, acinar cells and duct cells all contained weakly sulphated mucosubstances. Demilune cells contained neutral mucosubstances. Occasional demilune cells contained weakly sulphated mucosubstances. Carboxylated mucosubstances (sialomucins) were not found in the lingual glands.

Electron microscopic changes in human oesophageal epithelium in oesophagitis.

At the ultrastructural level the inflamed oesophageal epithelium shows a variety of lesions. These include mitochondrial damage, intracellular oedema, membrane whorls and dilation of the endoplasmic reticulum and the appearance of keratohyaline and parakeratosis granules. The basement membrane shows changes ranging from thickening to areas of loss of continuity. There is an increase in the anchoring fibrils. The spaces between the cells are enlarged and contain varying amounts of debris rich in neutral mucosubstances. There are less membrane-coated granules in the inflamed epithelium than in the normal. The intra-epithelial intrusive cells demonstrated were lymphocytes and neutrophil polymorphs.

Structure and carbohydrate histochemistry of the intestine in ten teleostean species.

The morphology and carbohydrate histochemistry of ten teleostean intestines are compared. Although there is an absence of regional differentiation seen in higher vertebrates, specializations in some species occur in the form of intestinal swellings, pyloric ceca and recta, the latter separated by a valve. The intestinal lumen is lined by a simple columnar epithelium interspersed with goblet cells; multicellular intestinal glands are absent. Thick basement membranes seen in centrarchids and Perca flavescens closely resemble the stratum compactum found in the lamina propria of esocids. Granular cells, which vary in number from species to species, are often seen in the mucosa and submucosa but less frequently in the muscularis. In species with intestino-rectal valves, a rectum is easily defined by the abrupt appearance of lower mucosal folds, more goblet cells and a thicker muscularis. In the remaining species the above features appear gradually in the distal intestine. Goblet cells show species variations in localization of epithelial mucosubstances, which in broad terms are recognized as sulfomucins, sialomucins and neutral mucosubstances. In both proximal and distal intestines the majority of goblet cells contain sialomucin although small amounts of sulfomucin are also often present. In species without intestino-rectal valves, no changes in carbohydrates occur between proximal and distal intestines. The possible significance of the heterogeneous character of digestive tract mucosubstances is discussed.

The forebody glands and surface features of the metacercariae and adults of Microphallus similis

Numerous unicellular glands are present in the forebody of the metacercariae and adults of Microphallus similis. The gland cell bodies lie beneath the tegumental perinuclear cytoplasm and ducts pass upwards penetrating the tegumental distal cytoplasm. Electron dense granules are secreted onto the forebody surface and into the oesophagus via small pits in the tegument. Histochemical tests revealed that the glands contain diastase-resistant neutral mucosubstances, RNA, protein, esterase and small amounts of acid phosphatase; inhibitor studies indicated that the esterase was acetylcholinesterase (EC 3.1.1.7). Ultracytochemistry showed that the mucosubstances were located in the general cytoplasm of the gland cells and that cholinesterase was present in the granules. The forebody of M. similis is covered by large toothed spines whereas the hindbody only bears short peg-like spines. It is suggested that the forebody spines may have an irritating, abrasive effect on the host's intestinal mucosa and that the secretion of acetylcholinesterase in the region of these spines may produce a ‘local anaesthetic’ effect on the host's gut by reducing movement in the immediate vicinity of the fluke, so decreasing the likelihood of expulsion.

Mucosubstance histochemistry of Brunner's glands, pyloric glands and duodenal goblet cells in the ferret.

The mucosubstance of Brunner's glands, pyloric glands and duodenal goblet cells were studied using the various histochemical methods. The secretions of both Brunner's and pyloric glands were similar in their histochemical reactions. They contained neutral mucosubstances as in these glands in man. The duodenal goblet cells showed variations in their histochemical characters. (i) The secretions of most of the deep cells and the majority of superficial cells contained sialidase-labile and sialidase-resistant sialomucins. (ii) There were a few superficial and occasional deep cells, the secretions of which contained sulphated mucosubstances. (iii) There were some goblet cells, more in the villi than in the crypts, the secretions of which contained a mixture of sialomucins and a sulphated mucin. The sialomucin was mostly sialidase-labile and partly sialidase-resistant.

Mucosaccharide histochemistry and histoenzymorphologic observations on the epidermis of Ariosoma balearicum de la Roche (Anguilliformes, Pisces)

The epidermis of Ariosoma balearicum consists of three layers - the basal layer, the middle layer and the outer layer. In between the basal cells are found clusters of small lymphocytes which show a moderat acid phosphatase activity. The middle layer and the outer layer are composed of three types of cells - the polygonal cells, the mucous cells and the club cells. The mucosubstances within the mucous cells exhibit the properties of neuraminic acid containing mucosaccharides with vicinal hydroxyl, sulfate esters and carboxyl groupings. The superficial cells of the outermost layer are capable of secreting sulphated and carboxylated mucosubstances to form an extracellular mucous coating which in fish epidermis seems to be necessary for the accumulation of electrolytes. The club cells are generally provided with one nucleus or double nuclei which appear displaced from the center and constitute the main histological component of the middle layer. The contents of these cells, in addition to a strong protein uptake visualized by the positive reactions exhibited in the coupled tetrazonium and mercuric bromophenol blue preparations, gave positive response to neutral mucosubstances which appear to be a glycoprotein involved in the slime secretions. Alkaline phosphatase and TPP-hydrolysing enzyme which could be detectable in the basal layer and the basement membrane were correlated with the probable role that they could play in the transportation of various chemicals and nutriments through the cell membranes.

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The normal female hamster respiratory epithelium at five airway levels was characterized with the use of coordinated morphologic and histochemical techniques. Five morphologic cell types were recognized in the trachea, stem bronchi, and primary bronchl: basal cells and neurosecretory cells that were basally located and did not reach the lumen and mucous cells [mucous goblet cells and small mucous granule cells (SMGC)], indifferent cells showing mucous-ciliary differentiation, and ciliated cells that reached the lumen. Two epithelial cell types were observed in the bronchioles, ciliated cells and nonciliated Clara cells, both of which reached the lumen. Mucous cells presented as either SMGC with a few small periodic acid-Schiff-positive granules (diastase-resistant neutral mucosubstances) or as goblet cells, filled with the same material. Mucous cells were columnar, and the cytoplasm was electron-dense and contained a well-developed endoplasmic reticulum and Golgi complex. The microvilli of the mucous cells were coated more thickly with colloidal iron than either the cilia or microvilli of ciliated cells. Approximately one-half the cells in the trachea, bronchi, and bronchioles were ciliated. Ciliated cells containing intracellular ciliated cysts with normal cilia projecting into a closed space or ciliated cells bearing compound cilia were observed infrequently. Neurosecretory cells were rarely observed. These cells contained characteristic dense-core granules.

Ultrastructural carbohydrate cytochemistry of gastric epithelium.

On examination with ultrastructural methods for visualizing the vicinal glycols and acid groups of complex carbohydrates, the most superficial surface epithelium of the rat gastric corpus displayed biphasic mucous droplets consisting of a cortex of hexose-rich (i.e. periodate-reactive) neutral mucosubstance and an uncharacterized denser core plus monophasic droplets with the neutral mucosubstance. In many surface epithelial cells of the foveolae, the biphasic and monophasic droplets with the neutral mucosubstance intermingled in varying proportions with monophasic droplets showing uniform periodate reactivity, a variable degree of dialyzed iron-binding--demonstrative of acidic glycoconjugate, and high iron--diamine affinity--demonstrative of sulphomucin. Deep foveolar epithelium displayed only monophasic droplets, most of which contained acidic periodate-reactive complex carbohydrate. Underlying cells, designated isthmus cells, exhibited monophasic or occasional biphasic granules containing sulphated, hexose-rich mucosubstance. Nascent droplets or granules near the Golgi zone differed from the mature organelles in the distribution of the glycoconjugate. Mucous neck cells occupied a deeper stratum and displayed a uniform population of monophasic mucous droplets with a loose meshwork of neutral mucosubstance. Techniques for demonstrating hexoses ultrastructurally stained all Golgi cisternae in the mucigenic epithelium, showing increasing reactivity toward the maturing face. Distinctive cisternae with moderate reactivity in the Golgi complex of isthmus cells were interpreted as GERL. Acidic mucosubstances were visualized only in the inner, mature cisternae of the Golgi complex of cells storing acidic glycoconjugates, and not in cisternae interpretable as GERL. The apical plasmalemma of isthmus cells uniquely exhibited abundant sulphated glycoconjugate and that of parietal cells revealed a less prominent, periodic neutral mucosubstance. Lateral and basal plasmalemmae varied from unstained to slightly reactive; basement membranes showed moderate reactivity with methods for visualizing complex carbohydrates. Abundance of glycogen further characterized surface epithelial cells of the corpus and of some parietal cells.

The ultrastructure and histochemistry of the cyst wall of Bucephalus haimeanus (Lacaze-Duthiers, 1854)

Metacercarial cysts of Bucephalus haimeanus occur throughout the liver of Pomatoschistus microps, greater concentrations being found around the gall bladder, blood vessels and periphery of the liver. The cyst wall is composed of three layers, an inner granular, middle vacuolated and outer nucleated layer. The inner layer consists of three zones, an inner zone of irregularly compacted, granular material; a middle densely compacted zone and an outer moderately compacted zone. Vacuoles of the middle layer vary in appearance and appear to be discharging their contents into the cyst cavity. The cells of the outer layer are flattened and elongated with elongate nuclei but no other distinct organelles. Occasionally at the outer edge of the cyst are electron-dense cells which are in the process of degeneration. Only small amounts of collagen and very few leucocytes are associated with the cysts.The cyst wall is carbohydrate in nature containing some free aldehyde groups, galactogen and neutral mucosubstances. The inner layer appears to consist of mainly acidic mucopolysubstances and weakly acidic sulphomucins. Large amounts of protein, carbohydrate-protein complexes and DNA are also present within the cyst wall. Alkaline and acid phosphatases, ATPase, β-glucuronidase and N-acetyl-β-glucosaminidase were detected. The origin and function of the cyst wall is discussed.

Spontaneous mammary carcinoma in C57BL and C3H mice: a histochemical study.

The mucosubstances in a series of 12 mammary carcinomas arising in C57BL mice were compared with those in tumours in nine C3H mice and with normal "resting" and lactating glands. A strain difference rather than a difference according to tumour type was found. The epithelial elements in the C57BL tumours contained mainly sialomucin with a little neutral mucosubstance and sulphomucin was present in some. Sulphated mucopolysaccharide was prominent in the stroma of these tumours. The C3H mice tumours contained no sulphomucin, less sialomucin but more neutral mucosubstance. The mucosubstances were studied in serial transplants of four tumours. An acid to neutral mucosubstance change with dedifferentiation of the tumour has been observed and found to be similar to a series of changes previously observed in a group of human gastrointestinal tumours.

Structure and carbohydrate histochemistry of the esophagus in ten teleostean species.

The histology and carbohydrate histochemistry of ten teleostean esophagi were compared. Structurally, the four layers of atypical vertebrate digestive tract were consistently present. The epithelium was always stratified and in all but one species (Ictalurus nebulosus) contained taste buds. Esophageal mucous cells were not the typical goblet cells seen in other vertebrates but appeared to be of six different types, pairs of which were associated with particular families. In esocids, poorly developed mucous acini and serous monogranular cells were present. In all species, the subepithelial connective tissue was not divided into definitive lamina propriae submucosae due to the absence of muscularis mucosae. Variably present in this connective tissue region were argentophilic fibers and in esocids only, randomly dispersed striated muscle fibers. The arrangement of the muscularis was reverse to that of the general vertebrate plan. In mucous cells, three general types of epithelial mucosubstances were identified and in broad terms were recognized as sulfomucins, sialomucins and neutral mucosubstances. Morphological differences were accompained by differences in carbohydrate localization, each esophageal epithelium containing at least two different mucosubstances. However, the mucosubstances identified in each mucous cell had a profile of characteristics different in some respects from any other. Thus teleostean esophagi appear to perform an integrated diversity of functions as reflected by their complex morphology and carbohydrate histochemistry.

Histochemical evidence of mucosubstances in the metaplastic epidermis and hair follicles produced in vitro in the presence of excess vitamin A

Pieces of upper lip and lower jaw from 13-day old mouse embryos grown in organ culture with excess vitamin A showed metaplastic changes in epidermis and hair follicles after 7 to 21 days. Changes were less marked in upper lip explants from 15-day old embryos. Areas of keratinizing epidermis showed a much higher incidence of Alcian blue-positive bodies when excess vitamin A was present. Histochemical tests with Alcian blue at critical electrolyte concentrations indicated moderately sulfated acidic mucosubstances in these bodies. Other patches of epidermis were transformed into stratified cuboidal epithelium producing PAS-positive, Alcian blue-negative, mucosubstances. The hair follicles, transformed by vitamin A into glands, developed three types of epithelium with apical granules, luminal borders and occasional goblet cells, all showing mucosubstances. Tests suggest that both metaplastic epidermis and glands secrete either neutral mucosubstances or non-sulfated acidic mucosubstances. Another response to vitamin A was a rapid loss of glycogen, particularly in hair follicles and the epidermal basal layer. It was concluded that excess vitamin A alters not only the morphogenesis but also the distribution and synthesis of polysaccharides in developing skin.

Histochemical studies on the mucins of the vertebrate tongues: XI. Histochemical analysis of mucosubstances in the lingual glands and taste buds of some birds

The tongues of six birds were investigated histochemically to determine the distribution and nature of mucosubstances in the lingual glands and taste buds. The anterior lingual glands in sparrow, kingfisher, parrot and pigeon hawk contained only one type of cells (seromucous) elaborating a mixture of neutral mucosubstances, sialomucins and sulfomucins. The anterior lingual glands in fowl and owl contained three distinct types of cells elaborating separately neutral mucosubstances, sialomucins and sulfomucins. The posterior lingual glands in most of the birds investigated except fowl contained only seromucous cells identical to the cells in the anterior lingual glands, while in fowl two distinct cell types were identified: one elaborating sulfomucins and the other sialomucins. Taste buds were found to be restricted in the glandular areas of the tongue in all the birds and always in close association with the gland ducts. Special adenomere cavities were seen only in the tongue of sparrow.