Histochemical features of the formation of the protective barrier of the stomach in various periods of ontogenesis of mammals

By means of a complex of histochemical methods it has been demonstrated that cytochemical differentiation of the tegmental epitheliocytes and secretory cells of the fundal glands takes place in different time. In Carnivores the tegmental epitheliocytes complete their cytochemical differentiation during the prenatal period, mucocytes--to the time of birth, and chief exocrinocytes--to the period of mixed feeding. In phytophagous animals formation of the protective barrier in the stomach occurs differently: in the mouse and hamster the cytochemical differentiation of the tegmental and glandular epitheliocytes is completed during the prenatal period, and in the rabbit and guinea pig--only by the 30th day after birth.     

Comparative lectin histochemical analysis of the duodenal glands in various mammals

Composition and histotopography of lectin receptors have been studied in 12 species of mammals with various nutritional specialization: carnivorous, phytophagous and omnivorous. In cells of the duodenal glands of the carnivorous and omnivorous receptors to concanavalin A and lentil lectin (D-mannosoglycans ) are absent and they are present in the glands of the phytophagous animals. In cells of some parts of the glands presence of receptors to soya bean lectin (N-acetyl-D-galactosamine++) is the most characteristic sign of the duodenal glands in the carnivorous and phytophagous animals. Together with certain differences, depending on the nutritional way of the animals, specific peculiarities of lectins binding with glandulocytes of the duodenal glands are demonstrated. The data on rearrangement of the lectin receptors are obtained during the process of cellular differentiation. Presence of N-acetyl-D-galactosamine++ remnants-biding soya bean lectin in composition of oligosaccharide++ chains of glycoconjugates is a sign of low differential degree of the glandular cells. In more differentiated cells concealment in oligosaccharide chains of D-galactose remnants (peanut and castor-oil lectins receptors) by L-fucose, N-acetil-D-glucosamin remnants and sialic acid can have place; this is demonstrated as accumulation of receptors to wheat germ and Laburnum anagyroides lectins in the glandular cells.     

Organization of the epithelial stratum of the mammary glandular tree

The main variants of epitheliocytes have been studied, cambial cells have been distinguished. Existence of peculiar columnar cells of epidermal proliferative units have been demonstrated in the mammary gland. They determine honeycomb-shaped organization of the epithelial stratum of the glandular tree of the organ.     

A cytomorphological analysis of the adhesion of fungi of the genus Candida to human and animal epithelial cells

Morphological pattern of adherence of Candida genus fungi to human and animal epithelial cells have been studied using histological and cytological methods. The interspecific differences of epithelial adhesive properties were observed: in one case, the mucous membrane epitheliocytes of different localization had different adhesive properties. Adhesive properties vary significantly within one type of epithelial cells. In individual epitheliocyte, the adherence of fungal blastospores was nonuniform: the greater density was observed in peripheral areas of cytoplasm.     

Ultrastructure of the epithelial cells of the duodenum at the early stages of experimental Escherichia infection

Ultrastructural changes of duodenal epitheliocytes were studied at the period from 15 minutes to 24 hours after inoculation using the model of experimental esherichiosis. The results obtained allow to determine the succession of ultrastructural changes and dynamics of adenylate cyclase activity of epithelial cells, involvement of endocrine cells in the pathological process. Combination of the certain morphological and cytochemical reactions and their dynamics allowed to make conclusions about typical ultrastructural changes in epitheliocytes at the early stages of experimental esherichiosis.     

Growth of fetal rat gastro-intestinal epithelial cells is region-specifically controlled by growth factors and substrata in primary culture

The mammalian gastro-intestinal tract can be divided into three parts: esophagus and forestomach, glandular stomach, and intestine. We have previously reported primary culture systems for duodenal and glandular stomach epithelial cells in which the cells express tissue-specific marker proteins. However, the effects of growth factors and substrata on cell growth have not been fully investigated. In this study a primary culture system was established for forestomach epithelial cells and the mechanism by which the growth of gastro-intestinal epithelial cells is controlled in primary culture was examined. Forestomach, glandular stomach and duodenal epithelial cells proliferated rapidly in culture, increasing their numbers about 30-, 20-and 10-fold, respectively, in the first 5 days. Scanning electron microscopy showed that these three types of epithelial cells exhibited region-specific morphologies in culture. Results on the effects of growth factors and substrata on the proliferation of the epithelial cells revealed that the culture conditions required to induce maximal epithelial growth differed. Forestomach and glandular stomach epithelial cells required similar combinations of growth factors to proliferate, and these were quite different from those required for duodenal epithelial cells. Glandular stomach and duodenal epithelial cells could proliferate in a serum-free condition while forestomach epithelial cells could not. Thus, glandular stomach epithelial cells exhibited intermediate characteristics between forestomach and duodenal epithelial cells regarding their growth factor requirement. Glandular stomach and duodenal epithelial cells could not proliferate on plastic without collagen substrata while forestomach epithelial cells could. Duodenal epithelial cells proliferated faster on collagen gels than on collagen films, and forestomach epithelial cells faster on collagen films than on collagen gels. Glandular stomach epithelial cells proliferated similarly on both substrata. Thus again, glandular stomach epithelial cells exhibited intermediate characteristics between forestomach and duodenal epithelial cells regarding their substratum dependency. We conclude that the growth of gastro-intestinal epithelial cells is affected by both growth factors and substrata, and that glandular stomach epithelial cells exhibit intermediate characteristics between forestomach and duodenal epithelial cells in responding to these factors. These results suggest that a head-to-tail gradient exists in the gastro-intestinal tract which controls the epithelial response to growth factors and substrata.     

Duodenal microanatomy of the domestic cat (Felis catus)

Duodenal samples were taken from similar locations in six cats, processed, stained, and examined via light microscope. There were no prominent circular folds (plicae circulares) or stratum compactum (lamina subglandularis). The 1072 microns x 201 microns villi were covered by 46 microns high columnar epitheliocytes proximally which decreased in height (41 microns) distally and displayed a 1.1-1.7 microns striated border. Globular leukocytes, mononuclear cells, and twenty-eight goblet cells (exocrinocytus calciformis) per villus were seen. The intestinal gland (crypt of Lieberkuhn) epithelium was 20 microns tall and had a less distinct striated border. The 515 microns simple straight tubular intestinal gland layer displayed distal branching. Many mitotic figures, 12 goblet cells per gland, and occasional columnar to triangular cells with red cytoplasmic granules were seen. The thickness of the lamina propria mucosa (glandular portion) decreased from proximal to distal (563-465 microns). The lamina muscularis mucosa had two layers and decreased in thickness distally (71-28 microns). The proximal muscularis mucosa was penetrated by the ducts of submucosal (Brunner's, duodenal) glands. The tela submucosa decreased in thickness distally (593-192 microns) and contained submucosal glands with 11.5-75 microns lumina for the first 1.5-2.5 cm. However, submucosal glands could be found to a distance of 8 cm. The glandular epithelium ranged from 7.5-22.5 microns in height. Only one type of secretory cell was observed, with both mucous and serous properties. The tunica muscularis ranged from 190-1425 microns (median thickness of 557 microns) and had two layers.     

Evidence of a microtrabecular cytoskeletal lattice in glandular cells of hydrozoan planulae

Hydrozoan planulae of Pennaria tiarella and Podocoryne carnea were processed for transmission electron microscopy using diethylene glycol distearate (DGD). The DGD functions as a removable embedding medium to produce embedment-free sections of intact planulae. Images of glandular cells obtained using embedment-free sections were compared with those from conventional Spurr-embedded sections. In unembedded sections a large number of thin anastomosing fibers were observed throughout the cytoplasm of the glandular cell. The fibers appeared to coalesce in certain areas to form thick bundles of fibers that partitioned the glandular cytoplasm into spherical compartments. The meshwork of fibers is three-dimensional and resembles a microtrabecular lattice. Mitochondria are suspended within and attached to the network of fibers, thus suggesting a cytoskeletal role of the fibers. This study documents the presence of a cytoplasmic fiber system within cells of intact invertebrate larvae.     

Effect of thyroid hormones on the histotopography of lectin receptors in the rat salivary gland

Using lectin-peroxidase technique, the influence of hypo- and hyperthyroidism on histotopography of glycoconjugates has been investigated in rat submandibular gland. The following lectins were used: peanut agglutinin (PNA), wheat germ agglutinin (WGA), Laburnum anagyroides lectin (LAL) and concanavalin A (con A). It has been demonstrated that hyperthyroidism is accompanied by the loss of con A, WGA and LAL receptor sites. Hypothyrodism enhanced con A binding to granular duct cells with a parallel reduction in WGA and LAL binding to these or other duct cells. Hypothyroidism as well as hyperthyroidism markedly enhanced PNA binding to duct epitheliocytes with redistribution of these lectin binding sites from the luminal surface of salivary ducts into the cytoplasm of duct cells. Possible interpretations of the observed phenomena are discussed.     

Localization of beta-glucuronidase in the kidney of the white rat after administration of the antidiuretic hormone

Distribution of beta-glucuronidase, one of three enzymes of hyaluronate hydrolases (HH) that hydrolyze extracellular glycosamine glycans (GAG), has been studied in the intact white rat kidney and under effect of antidiuretic hormone (ADH). Renal beta-glucuronidase is revealed as discrete granules in cytoplasm of epithelial cells, in the cortex and medulla and in the interstitial cells of the papilla. ADH increase for a short time in blood results in decreasing amount of granules in cytoplasm of epithelial cells of the collecting tubules, and a prolonged increase is accompanied with practically a complete disappearance of granules both in cytoplasm of epitheliocytes of the collecting tubules and in spindle-like interstitial cells. This effect is considered as a result of the enzyme discharge under ADH effect from the cells into the interstitial space--the place of the extracellular GAG position, HH action substrate. The possibility of the enzyme presence in the renal cells in the form of two fractions is discussed.     

Age-related changes in the ultrastructural organization of the human gingival epithelium

By means of transmissive and scanning electron microscopy 103 gingival bioptates in practically healthy persons at the age of 18-80 years have been studied. At ageing essential changes take place in all structural elements of the epithelium. The basal membrane is intermittent and loose. In cytoplasm of the cells of the basal layer epithelium the amount of microfilaments increases essentially, and as a result it becomes electron opaque. Tonofibrillar fasciculi of the spinous layer cells are fragmented, their contours are indistinct. In cytoplasm of the granular layer cells amount of keratohyalin granules increases, their size becomes large and their typical form is lost. In cytoplasm of the basal, spinous and granular layer cells the amount of organells decreases. Mitochondria acquire the appearance of electron translucent cavities with discomplexic, and sometimes, destroyed cristae. Rather great changes occur in intercellular interrelations. In all the layers some intercellular spaces are widen, in the spaces formed isolated desmosomes and other debries of cellular structures are formed. Sharp changes of microrelief of the granular layer epitheliocytes are observed. The ultrastructural rearrangements of epitheliocytes, revealed in the human gingiva, demonstrate certain disturbances in keratinization processes, in mechanical firmness, as well as in barrier function of the epithelial layer.     

Epitheliocytes of fundal glands and the relative volume of the parietal microflora of the stomach and small intestine mucosa in experimental chronic duodenal ulcer

The effect of experimental chronic duodenal ulcers and vagotomy on fundal gland epitheliocytes and membrane microflora has been studied in rats using light microscopy and stereometry on semithin sections. It is shown that in ulcers the relative amount of perietal and zymogen cells increases, while the volume of mucocytes decreases. Vagotomy leads to a decrease in the relative amount of parietal and zymogen cells and increases the relative amount of mucocytes. The relative volume of membrane microflora in gastric fundal and pyloroantral regions, duodenum and jejunum diminishes in ulcers and increases in vagotomy, as compared to the control.     

Quantitative characteristics of the endocrine cells of the duodenal glands of Carnivora

In the duodenal glands of the Carnivora investigated endocrine elements have been revealed, a part of them is presented as serotonin-producing EC-cells. Endocrine cells are situated in terminal parts and in glandular ducts, among them elements of open and close types are distinguished. Distribution of these cells in the glandular lobules is subjected to the distal gradient regularity, specific for the gastrointestinal tract mucosal membrane. Amount of endocrinocytes in the glands is much less than in the gut crypts. There is no correlation between distribution of the endocrine cells in the glands and in the crypts. The results of unifactor analysis of variance demonstrate a slight effect of the taxonomic position of the species on the number of endocrine cells in the duodenal glands. The proper endocrine apparatus of the duodenal glands is supposed to produce a local regulatory influence on the secretory activity of exogenic glandulocytes, as well as ensure humoral connections of the duodenal glands with other parts of the gastrointestinal tract.     

Morphological study of the development of the disseminate prostate in intact and castrated male pigs aged two to six months.

The light-microscopical and ultrastructural differentiation of the disseminate prostate was studied in 12 boars and 8 barrows, ranging in age from 8 to 28 and 12 to 27 weeks, respectively. The barrows had been castrated at the age of 8 weeks. In young boars the prostate is organized in an inner zone and an outer zone. The glandular tubules of the latter are more differentiated than those in the inner zone. Sexual maturation in the prostate is characterized by almost complete reduction of the inner zone and concomitant enlargement of the outer zone. This differentiation of the inner zone occurs progressively and is manifested by a higher density of the glandular tubules, by a decrease in the number of basal cells and by an increase in the secretory tubular cells containing more microvilli, organelles and secretory vesicles. A few weeks after castration, the characteristic morphology of the undifferentiated inner zone is found throughout the entire glandular layer. The number of glandular tubules in barrows has decreased, and their cells show minimal secretory activity, nuclear indentations and a decrease in their organelles. A further castration effect is vacuolization of the perinuclear cytoplasm in numerous glandular cells.     

Optico-structural machine analysis of the nuclear chromatin and cytoplasmic SH-groups of epitheliocytes of the oral mucosa of healthy subjects and in peptic ulcer of the stomach and duodenum

By means of optico-structural machine analysis, epitheliocytes of mucous membrane of the oral walls have been studied in 21 healthy persons and in 30 persons suffering from ulcerous disease of the stomach and duodenum. Changes in densitometric parameters of epitheliocytic nuclei at the ulcerous disease demonstrate increasing amount of heterochromatin and, hence, increasing degree of cell differentiation. Changes in contents and distribution of SH-groups in cell cytoplasm indicate that keratin is accumulated in it and that keratinization processes in the mucous membrane epithelium increase. In this way it is shown that the densitometric parameters are informative on distribution of hetero- and euchromatin in nuclei and SH-groups in epitheliocytic cytoplasm of the mucous membrane in the oral wall for estimation of morphofunctional state of these cells, normal and under pathological conditions.     

Cytologic features of premalignant glandular lesions in the upper gastrointestinal tract

The cytologic findings in 13 endoscopic brushing specimens from biopsy-proven premalignant glandular lesions (PGLs) of the upper gastrointestinal tract were reviewed retrospectively. The specimens were from ten patients: three with dysplasia in Barrett's esophagus, four with gastric adenomas and three with duodenal adenomas. One dysplasia in Barrett's esophagus and four adenomas (two gastric and two duodenal) had coexisting adenocarcinomas. Most pure PGLs were characterized cytologically by cohesive three-dimensional clusters of cells with more-or-less uniformly enlarged nuclei and an increased nuclear/cytoplasmic ratio. Crowding and molding were present within these clusters; however, the cells were arranged in a somewhat orderly or palisading fashion, instead of entirely haphazardly. In cases of carcinoma coexisting with adenoma or dysplasia, the atypical cells tended to be more pleomorphic and dyshesive. In one specimen from an adenocarcinoma arising in an adenoma, the adenomatous and carcinomatous components could be distinguished cytologically.     

Histochemical study of rabbit duodenal mucosa carried out using lectins

The Authors report histochemical findings about rabbit's duodenal mucosa. The present study has been carried out using five different lectins (Peanut Agglutinin (PNA), Dolichos Biflorus Agglutinin (DBA), Wheat Germ Agglutinin (WGA), Soybean Agglutinin (SBA), Ulex Europaeus Agglutinin I (UEA-I). These lectins have been labelled with Horseradish Peroxidase and binding sites have been stained with 3-3' Diaminobenzidine, according to Farragiana et al. The PNA reacted with the glandular cells, while the reaction was negative in the superficial cells. The DBA reacted exclusively with the glandular cells. The superficial and the glandular cells showed strong positive binding sites to the WGA and slight positive binding sites to the SBA. The UEA-I did not react with the epithelial cells. The presence of binding sites for the lectins we have used in the present study, shows a different glycoprotein composition of the cellular secretion, in comparison with the other animals we have already studied. In addition, these lectins can not be used as cellular differentiation markers in the epithelial cells of the rabbit's duodenal mucosa.     

Ultrastructural differentiation of glandular stomach (proventriculus) in chick embryo

Cytochemical characterization of mucosubstances of chick glanular stomach (proventriculus) changes from 15 days of development to postnatal and adult stages was studied. To corroborate these data cytochemical, ultrastructural and ultracytochemical study of chick embryo proventriculus from 7 to 20 days of development was performed. At the 7th day several layers of undifferentiated cells formed an epithelium which covered the walls of the glandular stomach. Mocosubstances were not found. Between the 9th and 5th day a single layer of cylindrical cells was encountered forming invaginations which originated deep glands. Three types of cells were separated from the above mentioned layer, dark, clear and undifferentiated. The dark cells had organelles which are involved in protein synthesis and the clear ones were rich in mitochondria. Argentaffine cells appeared at 15th day instead mucosubstances formed a thin coat on the epithelium at 9th day which increased at the end of development in the apical cytoplasm and gland cells. These observations demonstrate that proventriculus of chick embryo has ultrastructurally differentiated cells involved with enzymatic and hydrochloric acid secretion after the 9th day. These progressive events are correlated with the digestion process of yolk during embryogenesis. At the end of development the proventriculus has completely organized the glandular layer.     

Bipolarity of duodenal enterochromaffin cells in the rat

Summary Enterochromaffin cells of the rat duodenum have been studied immunocytochemically by use of a specific antiserum to serotonin. At the light-microscopic level serotonin immunoreactivity was observed in enterochromaffin cells located in the epithelium of the duodenal mucosa. Most of the serotonin-immunoreactive material was localized to the basal portion of the enterochromaffin cells, but small amounts of immunoreactive material were regularly observed in the apical portion. At the electron-microscopic level serotonin immunoreactivity in enterochromaffin cells was found to be concentrated over the dense cores of the cytoplasmic granules. The majority of these granules was located in the basal cytoplasm of the enterochromaffin cells, but serotonin-immunoreactive granules were also observed in the apical cytoplasm immediately beneath the microvilli. These observations indicate that duodenal enterochromaffin cells are bipolar and that they secrete serotonin both basally, to the circulation, and apically, to the gut lumen. Rat duodenal enterochromaffin cells thus appear to have an exocrine as well as an endocrine function.