Morphofunctional changes in the chief cells of the intrinsic glands of the stomach in the presence of an excess of glucocorticoid and sex hormones in the body

In 260 male white rats electron microscopically, cytochemically with biochemical estimation of proteolitic activity of gastric juice, it has been demonstrated that principle cells of the gastric glands, after injection of small doses of hydrocorticone, respond with a complex of adequate ultrastructural, cytochemical and functional changes directed to realization of the drugs stimulating effect. With increasing time of the experiment, or the doses, progressively developing distrophic processes, involving predominantly the protein-synthesizing apparatus and, to a less degree, the mitochondrial mechanism, are observed in the principle cells. This results in inhibition of the pepsinogene synthesis phase and in decreasing proteolytic activity of gastric juice. Reaction of the principle cells is independent on testosterone doses; the protein-synthesizing and mitochondrial (to a less degree) apparatuses develop; the pepsinogene extrusion phase is suppressed; it results in its deposition in cytoplasm and in decreasing proteolytic activity of gastric juice. When testosterone and hydrocortisone are injected simultaneously in small doses, the stimulating effect of hydrocortisone to the principle cells is abolished and at large doses it is sharply manifested and is not accompanied with destructive changes in the principle cells.     

Characteristics of gastric secretion in suckling calves.

In the investigations on five calves with fistulae in the abomasum and oesophagus the secretion of gastric juice as well as its content of proteolytic enzymes and hydrogen ions were measured. It was found that the gastric juice of calves possessed a fairly high proteolytic activity (about 6 UP/l) and hydrogen ion level already on the 3rd day of life. Sham feeding produced two different types of gastric secretion response, depending on the age of calves. In the first week of life sham feeding caused an inhibition of secretion which became more intensive only about 30 min following food administration. In the second week of life gastric juice secretion increased immediately after feeding.     

Ultrastructure of cells of proper gastric glands and their stroma in hemorrhagic shock in the rat.

Ultrastructural changes in the cells of the proper gastric glands and their stroma are biphasic in the hemorrhagic shock. First phase: "paralysis" of the capillary vessels with an oedema of their stroma, an intracellular oedema with hydropic degeneration of the parietal cells and a degranulation of the argentaffin and argentaffin-like cells. Second phase: ischemia of the gastric mucosa with ultrastructural features of: a) the increased secretion of the parietal cells, b) the degranulation of the chief gastric cells, c) the increased secretion processes in the mucous neck cells. These findings suggest that at last 2 factors coincide in the pathogenesis of so-called stress gastric ulcerations: biphasic disturbances of blood circulation initially damage the gastric mucosa and are followed the digestive hyperactivity of the gastric juice finally resulting stress ulceration.     

Immunocytolocalization of human gastric lipase in chief cells of the fundic mucosa

Summary The presence in human gastric juice of a lipase secreted by the gastric mucosae has been reported previously, but its exact cellular origin has not yet been established. Polyclonal antibodies specific to human gastric lipase (HGL) were prepared, and used by an immunofluorescence technique to label cells producing HGL. This immunocytolocalization was correlated with that of pepsin (chief cells) and parietal cells using specific polyclonal or monoclonal antibodies.Our results clearly establish that HGL is exclusively located in the chief cells of fundic mucosa; furthermore, it was found to be always co-located with pepsin. No HGL was observed in the parietal or mucus cells. HGL was always detected intracellularly, either in secretory granules of the apical region of the chief cells, or revealed by more diffuse cytoplasmic labelling.Abbreviations HGL Human gastric lipase - SDS PAGE Sodium dodecyl sulfate-polyAcrylamid gel electrophoresis - PBS Phosphate buffer saline     

Immunocytolocalization of human gastric lipase in chief cells of the fundic mucosa

The presence in human gastric juice of a lipase secreted by the gastric mucosae has been reported previously, but its exact cellular origin has not yet been established. Polyclonal antibodies specific to human gastric lipase (HGL) were prepared, and used by an immunofluorescence technique to label cells producing HGL. This immunocytolocalization was correlated with that of pepsin (chief cells) and parietal cells using specific polyclonal or monoclonal antibodies. Our results clearly establish that HGL is exclusively located in the chief cells of fundic mucosa; furthermore, it was found to be always co-located with pepsin. No HGL was observed in the parietal or mucus cells. HGL was always detected intracellularly, either in secretory granules of the apical region of the chief cells, or revealed by more diffuse cytoplasmic labelling.     

Compensation of exocrine pancreatic insufficiency during its partial, subtotal and total resection

Chronic experiments on dogs have shown that the damage of extra-secretory pancreatic function by duct ligation caused marked compensatory changes of stomach function. The increase in gastric juice secretion and gastric juice proteolytic activity was accompanied by the reduction in its acidity. In addition to quantitative changes, qualitative shifts were revealed (amylolytic activity in strongly acid pH-reaction), never observed in the gastric juice of intact animals. Partial pancreas resection (up to 75%) both in control and test animals 10-14 months after pancreatic duct ligation was not accompanied by significant changes in gastric juice secretion. Total pancreas resection in dogs with previous pancreatic duct ligation caused neither prompt animal death, as in the control, nor the inhibition of compensatory reactions of gastric juice secretion.     

Effects of Gastric Freezing on the Isolated Denervated Gastric Pouch in Dogs

The purpose of this study was to investigate the effects of gastric freezing on the chief and parietal cells of the stomach. To carry this out six Heidenhain pouch dogs were prepared. The secretions of these pouches were studied, both for HCl levels and pepsin activity, for 30 days before and 30 days after the freezing of the pouch. Freezing was carried out for one hour at -17 to -20° C. The results show that no essential depression of either the HCl levels or the pepsin activity of the juice was effected by freezing. Therefore, it is deduced that freezing does not affect the functional activity of either the chief or the parietal cells. Complications such as ulceration and fistula formation into adjacent organs occurred as a result of freezing in certain pouch dogs.     

Morphocytochemical reaction of the main exocrinocytes of the proper gastric glands to orchiectomy

In the experiment performed on 52 white male rats by means of light and electron microscopy, cytochemistry and biochemistry, it has been stated that bilateral removal of the testes inhibits proliferation of epitheliocytes in the collum of the proper gastric glands and their differentiation into the main exocrinocytes. This results in decreasing amount of the latter and in development of certain destructive processes in them. They concern mainly the protein-synthesizing apparatus and to a less degree the mitochondrial and lysosomal apparatus. RNA content, ATPase activity oxidoreductase of Krebs cycle and pentose-phosphate shunt decreases, and that of glycolysis increase. Pepsinogen synthesis and its extrusion are inhibited; this is accompanied with a decreased proteolytic activity of the gastric juice. Regenerative processes in the main exocrinocytes are also inhibited.     

Quantitative changes in gastric juice proteases in dogs during ulcer formation in the stomach]

Disc-electrophoretic investigation of proteases of the gastric juice of dogs under normal conditions and in experimental atophan ulcer of the stomach demonstrated that in fasting dogs ulceration was accompanied by marked quantitative changes in the protease spectrum. Fraction No. 6 showed a significant elevation against the background of total reduction of proteolytic fraction of the gastric juice in sham feeding of ulcerated dogs. Sham feeding replete ulcerated dogs indicated a marked increase of fractions No.1 and No.2. The noted changes of the gastric juice protease spectrum may be of ulcerogenic significance.     

Restoration of secretory activity of digestive glands in conditions of acute hyperkinesis at persons with different levels of motor activity

The dynamics' features of restoration reactions of the secretory function of gastric glands have been studied at patients with differences in the level and specificity of daily physical activity. The dependence between the level and characteristics of daily physical activity and reactivity of the secretory apparatus of the gastric glands in the recovery period after the muscle load has been established. The high reactivity of the secretory activity of digestive glands is typical for individuals with high level of daily physical activity. The differences of the functional stability of the secretion's mechanisms of various components of gastric juice under the influence of physical exercise and in recovery have been revealed. The greatest stability of the secretory mechanisms of digestive glands has been discovered at athletes practicing with the development of endurance--at skiers. Heterochronous reducing reactions of gastric and pancreatic secretion after physical stress have been defined. The reduction of secretion's mechanisms of gastric juice ingredients and the electrolyte and acid composition of digestive juices isn't simultaneous: the first order is for ferment's secretion. The inverse relationship between the content of digestive enzymes pepsinogen-1 and -2 in blood's serum and the concentration of proteolytic enzymes in the gastric content has been found at persons with various degrees of adaptation to the muscular tension.     

Ultracytochemical study of oxidoreductases in the parietal cells of the gastric mucosa in gastric cancer

Ultracytochemistry was used to study and compare cytochromooxidase, succinate dehydrogenase and NADH-dehydrogenase activity in gastric mucosa parietal cells in health and in gastric carcinoma associated with decreased acidity of gastric juice. The study demonstrated the reduced activity of the enzymes listed in the mucosal parietal cells in gastric carcinoma. This finding is interpreted as a consequence of disturbed energy supply of hydrochloric acid secretion in gastric carcinoma.     

Cellular and subcellular localization of progastricsin in calf fundic mucosa: colocalization with pepsinogen and prochymosin

The presence of gastricsin in bovine abomasal juice has been reported previously, but its exact site of origin has not yet been established. Specific polyclonal antibodies were used in the peroxidase-antiperoxidase method or the protein A/gold technique to label cells producing progastricsin. This immunocytolocalization was correlated with that of pepsinogen and prochymosin using specific polyclonal antibodies against those zymogens. The present study clearly established that progastricsin was located exclusively in chief, mucous neck, transitional mucous neck/chief, foveolar epithelial and surface epithelial cells of the calf fundic mucosa. Furthermore, progastricsin was found to be colocalized with pepsinogen and prochymosin in the same secretory granules of these cells. Progastricsin was not observed in parietal, gastric endocrine and undifferentiated neck cells.     

CRYSTALLINE PEPSIN : V. ISOLATION OF CRYSTALLINE PEPSIN FROM BOVINE GASTRIC JUICE

1. A method has been described for isolating a crystalline protein with high proteolytic activity from bovine gastric juice by means of precipitation with magnesium sulfate and fractionation of the precipitate with acetone and magnesium sulfate. 2. The crystalline protein obtained in this way has the same crystalline form, optical activity, and specific activity, as determined by a number of methods, as does the crystalline protein previously isolated from swine gastric mucosa. 3. The solubility of the two preparations, however, is additive so that they are different although very closely related proteins.     

Immunocytochemical localization of polyamines in the gastrointestinal tracts of rats and mice

An immunocytochemical method using a recently produced monoclonal antibody (ASPM-29) with an antibody specificity to spermine (Spm) and spermidine (Spd) fixed in situ, was used to demonstrate an immunocytochemical localization of polyamine (PA) pools in the gastrointestinal tracts of rats and mice. High PA immunoreactivity was always found in the cytoplasm of cells not only at the cell proliferative zone or the precursor cell zone but also at the neighboring non-proliferative premature cell zone of the epithelium, and a gradient of decreasing PA levels was noticed from these cells to the fully mature differentiated gastric surface mucous cells and absorptive cells of the small and large intestines. Also, strong staining for PAs was seen in the cytoplasm of fully differentiated gastric chief cells and neurons of both the myenteric and submucous plexuses, whereas the nuclei of the cells remained virtually unstained. These results may suggest that PAs are closely associated with the high biosynthetic activity in the cells of the gastrointestinal mucosa of normal rats and mice. This seems to be consistent with the PA imunocytochemical results previously obtained for neoplastic cells and active protein- or peptide-secreting cells, including exocrine or endocrine cell types.     

Participation of protein kinase C in hormonal regulation of pepsin secretion in the rat stomach]

It was shown that pentagastrin (0.5 micrograms/100 g of body mass) increases the activity of Ca2+ and phospholipid-dependent protein kinase C in the membrane fraction of rat gastric mucosa cells. This effect of pentagastrin is accompanied by a decrease of the protein kinase C activity in the cytosolic fraction. Chromatography of the membrane fraction revealed an additional peak of the enzyme activity. Analysis of isolated gastric mucosa cells demonstrated that pentagastrin (10(-8)-10(-6) M) (but not 10(-4) M histamine) added to the incubation mixture increased the protein kinase C concentration in the membranes. The pentagastrin effect was directly correlated with the amount of pepsin-producing chief cells in the cellular pools. Carbacholine, another well-known pepsin secretion stimulator, was able to activate, similar to pentagastrin, the protein kinase C activity. It is concluded that protein kinase C plays a prominent role in hormonal regulation of the chief gastric cell function.     

Characterisation of the barrier caused by luminally secreted gastro-intestinal proteolytic enzymes for two novel cystine-knot microproteins

It was the aim of this study to evaluate the stability of two novel cystine-knot microproteins (CKM) SE-ET-TP-020 and SE-MC-TR-020 with potential clinical relevance towards luminally secreted proteases of the gastrointestinal tract in order to gain information about their potential for oral administration. Therefore, the stability of the two CKM and the model-drug insulin towards collected porcine gastric and small intestinal juice as well as towards isolated proteolytic enzymes was evaluated under physiological conditions. No intact SE-ET-EP-020 was detected after few seconds of incubation with porcine small intestinal juice. SE-ET-TP-020 was also degraded in porcine gastric juice. Furthermore, SE-ET-TP-020 was extensively degraded by isolated chymotrypsin, trypsin and pepsin. Moreover, it was degraded by elastase. SE-MC-TR-020 was degraded entirely within approximately 2 h when incubated in porcine small intestinal juice, whereas no degradation was observed within a 3 h incubation period with porcine gastric juice. In presence of the isolated proteolytic enzymes, SE-MC-TR-020 was only slightly degraded by trypsin and pepsin, whereas elastase caused no degradation to SE-MC-TR-020 at all. Chymotrypsin was the protease that caused most degradation to SE-MC-TR-020. The model drug insulin was degraded extensively by chymotrypsin, elastase, pepsin and trypsin as well as by porcine gastric and porcine small intestinal juice. In conclusion, a precise characterisation of SE-ET-TP-020 and SE-MC-TR-020 degrading luminally secreted GI enzymes has been made, which is an important and substantial prerequisite for the further optimisation of these CKM.     

Proliferative reaction of the epithelium of gastric glands to vagotomy and sympathectomy

By means of autoradiography and metaphase arrest technique 24-hour rhythms and intensity of proliferative processes in the epithelium of gastric glands were studied, as well as morphometric status of these structures after vagotomy and drug sympathectomy in the rat. In both cases the reaction of glandular epithelium appeared with the increase of mitotic activity and the decrease of cell synchronization before the entry into DNA--synthesis and mitosis. The number of epithelial cells in gastric glands became reduced, but after vagotomy, unlike sympathectomy, mainly because of the lack of chief cells. Thus, the trophic control upon the dynamic structure of gastric glands may have different mechanisms of realization for parasympathetic and sympathetic nervous system, probably, through the influence on cell proliferation or differentiation.     

Morphofunctional analysis of the chief (peptic) glandulocytes in the stomach of hibernating rodents

Ultrastructure of the chief gastric gland cells has been studied in the ground squirrel (Cytellus erythrogenys Brandt) at various seasons of the year. When the ground squirrel is in the active state, the cells studied do not differ from those in other animals. During winter hibernation, their activity is inhibited. During first days of awakening, the activity of the chief cells increases; the protein-synthesizing apparatus, the Golgi complex become activated, secreating granules accumulate. At the same time, the greatest number of the destroing cellular elements and discharge of the residual bodies out of the activizing cells are observed. Restoration of the normal structure in the chief cells is completed by the end of the second week. No signs demonstrating transformation of additional cells into the chief ones are revealed. Non-differentiated cells in the ground squirrel fundal glands are widely distributed along the whole length of the glandular cervix and occur even in the upper parts of its body.     

Superoxide dismutase activity of digestive secretions

A superoxide dismutase (SOD) activity in saliva and gastric juice has been revealed using the NBT reduction and adrenaline autooxidation assays. This activity in saliva is induced mainly by enzymic mechanisms, whereas in gastric juice--by nonenzymic mechanisms. The gel chromatographic analysis of saliva has demonstrated the presence of several SOD isoenzymes. Nonenzymic mechanisms of the SOD activity in gastric juice involve the ionic activity of H+, Cu2+ and that of reduced glutathione. The SOD activity in gastric juice can be decreased by adding antacids and Fe ions. The effect of Fe3+ is found to be different depending on the methods of the SOD activity evaluation. Such metal-chelating agents as histidine and organic acids increase the SOD activity in gastric juice.     

Isolation and purification of proteolytic enzymes on organo-silica sorbents with immobilized gramicidin S

Biospecific sorbents for affinity chromatography of proteolytic enzymes have been synthesized by attaching cyclopeptide antibiotic gramicidin S to organo-silica supports. It is shown possible to attach gramicidin S to the organo-silica supports using glutaric aldehyde, p-benzoquinone, soluble and insoluble carbodiimides. The sorbents prepared by these methods were successfully applied for the purification of the crude pepsin from horse gastric juice and proteolytic complex produced by Acremonium chrysogenum.