Streptozotocin-induced alterations in rat liver Golgi complexes are ameliorated by BMOV [Bis(maltolato)oxovanadium(IV)] activity.

Twenty years ago, we detected the interdependence between structure and function of rat liver Golgi complexes that are characteristic for streptozotocin diabetes, which served us in further investigations as a useful indicator of the effectiveness of drugs we were testing. This work presented results obtained in eight groups of rats (four control and four diabetic) that were administered orally either bis(maltolato)oxovanadium(IV) [BMOV] or maltol alone. The activities of the rat liver Golgi marker enzyme, galactosyltransferase [GalT], as well as the morphology of Golgi complexes were studied in situ using an electron microscope; parallel estimations of vanadium concentration and phospholipid percentage were made in Golgi-rich preparations isolated from the liver. Our main findings were normalization in diabetic animals orally treated with 1.8 mmol BMOV in 0.09 mol NaCl solutions over seven days, which demonstrated an accompanying increase in phosphatidic acid (PA) percentage (p < 0.05) compared to controls. In the diabetic groups, Pearson's test showed a positive double correlation between GalT activity, vanadium concentration, and PA percentage in Golgi-rich membrane preparations from the liver. Additionally, a negative correlation was found between vanadium concentration and phosphatidylcholine percentage in the fractions.

The influence of a new vanadium compound, bis(2,2'-bipyridine)oxovanadium(IV) sulphate on liver golgi complexes from control and streptozotocin-diabetic rats.

Among the previously studied organic vanadium derivatives showing an anti-diabetic action, we investigated a new complex, bis(2,2'-bipyridine)oxovanadium(IV) sulphate. We tested its ability to normalise parameters previously described for streptozotocin (STZ)-diabetes, such as lower yields of Golgi-rich membrane fraction isolation, decreased activity of Golgi membrane marker enzyme - galactosyltransferase (GalT) - and altered morphology of rat liver Golgi complexes. Oral application as a drinking solution of 1.8 mmol bis(2,2'-bipyridine)oxovanadium(IV) (dissolved in 0.09 M NaCl) caused a similar dispersion of GalT activities in both vanadium treated groups, control and diabetic. Very low activities of the enzyme (characteristic for untreated diabetes) we found only in approximately 35 % of the STZ-diabetic rats treated with the new vanadium compound. The morphology of liver Golgi complexes in diabetic rats treated with bis(2,2'-bipyridine)oxovanadium(IV) sulphate was improved, which manifested itself in the reappearance of vacuoles with VLDL and coated and uncoated secretory vesicles. In view of biochemical and morphological parameters of normalised diabetic rat liver Golgi apparatus, the new vanadium complex was more effective than bis(oxalato)oxovanadium(IV) or bis(kojato)oxovanadium(IV), but in our experimental model, the best anti-diabetic, orally applied drug was the bis(maltolato)oxovanadium(IV) previously investigated.

Biochemical and morphological alterations in rat liver Golgi complexes after treatment with bis(maltolato)oxovanadium(IV) [BMOV] or maltol alone.

Oral treatment with maltol or bis(maltolato)oxovanadium(IV) [BMOV] alters the biochemical activity of the rat liver Golgi marker enzyme, i.e., galactosyltransferase (GalT), and the organelle morphology in a relatively short time. Four groups of rats were investigated: control (C), treated with BMOV for 2 days (pVC), treated with BMOV for 7 days (C+V), and treated with maltol alone for 7 days (C+M). All drugs were administered as drinking solutions. These conditions were used, because normalization of galactosyltransferase activity (GalT) and morphology of rat liver Golgi complexes were previously found by us in streptozotocin-induced diabetes. In this paper, we present the influence of BMOV or maltol alone (as a vanadium ligand in BMOV compound) on rat liver Golgi complexes. The lowest statistically significant enzyme activity, in comparison with three other groups of rats (p < 0.01), was found in rats treated with BMOV solution for two days (pVC). Liver Golgi complexes in these rats showed relatively slight changes as compared with controls. The activity of GalT was similar to controls of the C+V and C+M groups. Morphological examinations of the Golgi apparatus in rats treated with vanadium salts revealed a slightly increased secretory activity. In response to various agents used in experiments, the Golgi complexes were generally reduced in size, except for the (C+M) group. Not only cisternae, but also vacuoles and associated vesicles on both sides of stacks were reduced in almost all Golgi structures. Ultrastructural findings were generally in agreement (except for pVC group) with biochemical results (yields of liver Golgi-rich fractions, activity of galactosyltransferase) obtained in the same rats.

Biochemical and morphological study on liver Golgi complex in streptozotocin-diabetic and control rats treated with bis(kojato)oxovanadium(IV) [VO(ka)2]x2H2O. Part I. One week treatment with vanadium compound.

In comparison with untreated control, reduced body and liver weights were found in two groups of rats (such as control and STZ-diabetic) treated orally with bis(kojato)oxovanadium(IV) solution. Free blood sugar in STZ-diabetic rats was lower by about 38%, but did not achieve euglycemic values. Yields of Golgi-rich fraction were lower than those in untreated controls, similar to the activity of galactosyl transferase (GalT) in both vanadium treated groups (control and diabetic). Under electron microscope in the control kojate-treated group, subcellular changes were observed. The morphology of Golgi apparatus was typical, resembled that in untreated animals. In diabetic animals treated with kojate subcellular changes were less severe. Golgi apparatus was usually semicircular or arched in shape similar to that observed previously in diabetic untreated rats.

Biochemical and morphological study on liver Golgi complex in streptozotocin-diabetic and control rats treated with bis(kojato)oxovanadium(IV) [VO(ka)2]x2H2O. Part II. Prolonged treatmentwith vanadium compound.

In this study oral treatment with bis(kojato)oxovanadium(IV) solution was administered twice. The first, short vanadium treatment (so called pretreatment) was used to accustom animals to the flavour of this liquid. After 2-2.5 weeks the second treatment, in high concentration and for a longer time served as a "drug". The physiological parameters such as weights of animals and their livers, reduction of liquid and food intake were lower than in control. Activity of Golgi marker enzyme GalT was significantly lower in both vanadium treated groups (p < 0.01), but in diabetic vanadium treated group it was higher, albeit not significantly, than in control vanadium treated rats. The morphology of Golgi complexes in diabetic rats on prolonged vanadium treatment was similar to that in the one week treated rats (see Part I). Rounded stacks of cisternae characteristic of untreated streptozotocin (STZ)-diabetes were also seen in vanadium treated diabetic rats, but in comparison with untreated diabetic livers, the secretory activities of Golgi complexes were preserved or even stimulated.

[Familial LCAT deficiency--clinical picture. Case report]. / Rodzinny niedobór LCAT--obraz kliniczny. Opis przypadku.

We report a case of familial LCAT deficiency, diagnosed in a 35 year old women. The disease manifested itself by a presence of proteinuria, corneal opacities and haemolytic anaemia with target cells. Suspecion of familial LCAT deficiency was based on renal biopsy, which revealed characteristic serpiginous fibrillar deposits in electron microscopy. The diagnosis was confirmed by a marked decrease of estrified cholesterol, low HDL-cholesterol concentration, decrease of LCAT activity in serum, typical "stacked coins" picture of HDL lipoproteins in electron microscopy examination and positive familial history--diagnosis of LCAT deficiency in dialysed brother of patient.

Fabry's disease. Report of the case diagnosed on the basis of routine ultrastructural examination of the renal biopsy.

A 45-year-old man who developed proteinuria was diagnosed as having Fabry's disease on the basis of renal histological findings and prominent decreases in alpha-galactosidase A activity in blood leukocytes.

Influence of bis(maltolato)oxovanadium(IV) on activity of galactosyltransferase (GalT) and morphology of rat liver Golgi apparatus in control and streptozotocin diabetes.

The relation between bis(maltolato)oxovanadium(IV) (BMOV) influencing the biochemical activity of rat liver Golgi apparatus and the morphology of this organelle was studied in normal and streptozotocin-diabetic rat livers. Ultrastructural examinations revealed marked differences in the morphology of Golgi apparatus in three groups of animals. In the control rats treated only with 0.5% NaCl we did not find any biochemical and morphological changes. Marked changes were found in the rat liver after 1.8 mmol BMOV in 0.5% NaCl (as drinking solution) applied for 7 days, so-called "control" group for vanadium. In this group Golgi apparatus seemed shorter than in the diabetic animals. Finally, the same treatment of rats with previously induced SZ-diabetes, showed relatively small morphological alterations. The ultrastructural observation was compatible with the activity of galactosyltransferase (GalT), the Golgi marker enzyme. In diabetic rats treated with BMOV the activity of this enzyme was almost the same as in controls. Summing up dramatic alterations, previously found in diabetic-untreated rats [22], normalized after orally applied BMOV solution, even after a short time.

ECL cells involvement in the isoproterenol-induced gastroprotection. An ultrastructural study.

The central and peripheral adrenergic systems are involved in the regulation of several functions in the gut including the maintenance of gastric microcirculation and gastric secretion but little is known about the role of the adrenergic system, in particular, beta-adrenoceptors in the phenomenon of gastroprotection. In this study acute gastric lesions were provoked by an intragastric (i.g.) application of 100% ethanol in rats with topical application of isoproterenol (ISO) (1 mg/kg) or subcutaneous (s.c.) administration of ranitidine (RAN) (40 mg/kg) or both. An area of gastric lesions was determined by planimetry, gastric blood flow (GBF) was determined by H2-gas clearance technique and gastric specimens were taken for histology and electron microscopy. It was found that ISO reduced ethanol-induced gastric mucosal lesions and this effect was accompanied by a rise in GBF. In contrast, RAN applied s.c. in a dose that produced almost complete achlorhydria, failed to affect ethanol-lesions but attenuated significantly ISO induced gastroprotection and abolished the increase in the GBF induced by this beta-adrenoceptor agonist. Histology and ultrastructural study revealed that pretreatment with ISO influenced HCl production reflected by the elaborated secretory surface of parietal cell intracellular canaliculi. All these changes were accompanied by the ultrastructural changes in Golgi apparatus in ECL-like cells (histamine storing cells). Pretreatment with ISO caused the collapse of Golgi profiles and only peripheral sacs were not compressed but such a change was not observed after RAN treatment. The best developed Golgi was, however, seen in the control rats without any treatment. Secretory granules in ECL cells were significantly expanded after pretreatment with ISO but did not show significant morphological changes in rats pretreated with RAN. We conclude that 1. ISO protects the gastric mucosa injured by ethanol presumably due to increased gastric microcirculation, and 2. ECL-like cells are actively involved in ISO-induced gastroprotection possibly by the increased histamine release which is reflected by ECL-cell and Golgi apparatus morphology.

Simvastatin-induced myopathy in a patient treated for hypercholesterolemia. Morphological aspects.

Functional and histological changes in skeletal muscle developing during hypolipemic therapy, especially with 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase inhibitors are rare. This paper reports a case of simvastatin-induced myopathy confirmed histopathologically and ultrastructurally.

Influence of LEPK on biochemical activity and morphology in situ of liver Golgi apparatus from control and streptozotocin-diabetic rats.

This paper presents yields of Golgi-rich membrane isolation, the activity of galactosyltransferase (GalT), the marker enzyme of Golgi apparatus as well as the morphology of the organelle from the livers in situ, in two groups of rats. One group consisted of control rats injected twice intraperitoneally with LEPK. Second group consisted of rats injected with LEPK and additionally after 24hrs given streptozotocin (SZ) to induce experimental diabetes. The results were compared with our previous investigations in control and diabetic rats. In the latter the activity of GalT was diminished, therefore diminishing glycosylation ability, and destructing Golgi apparatus morphology. This experiment shows that two-fold injection of LEPK prior to SZ does not prevent from changes in such biochemical parameters as free blood glucose level, yield of liver Golgi membranes isolation or total activity of GalT. For the first time in c. 30% of investigated rats the inactive enzyme of Golgi apparatus was found in the rats treated with LEPK+SZ. Morphological investigations of liver Golgi apparatus in rats treated with LEPK show slightly increased secretory activity with similar to untreated control rats morphological structure of this organelle. In the rats treated with LEPK and SZ the same morphological changes as in diabetic liver were found, however, such dramatic alterations as in SZ-diabetic rats were never found, irrespective of active or inactive GalT.

The use of congo red as a lyotropic liquid crystal to carry stains in a model immunotargeting system--microscopic studies.

The lyotropic liquid crystal dye-Congo Red was used as a carrier in a model immunotargeting system constructed from sheep red blood cells (SRBC) representing the antigen target and rabbit IgG anti-SRBC as the specific driving immunoglobulin. Rhodamine B and Hemin stains were chosen as example chemicals carried to the target. The carried stains were introduced to the micellar organization of Congo Red by intercalation. Preserving its supramolecular organization, Congo Red binds spontaneously and selectively to antibodies that have altered structure extorted by interaction with the antigen in the immune complex. The functionality of the studied immunotargeting model was verified by fluorescence and electron microscopy. The results indicate that the supramolecular nature of protein ligands offers new ligation capabilities possibly useful for carrying stains or drugs in immune-oriented systems.

Myocardial damage in thyrotoxicosis--ultrastructural studies.

Lack of uniform views concerning subcellular image and possible meaning of free radicals in the myocardial damage prompted us to study rat hearts submitted for chronic thyrotoxicosis. The studies were performed on 30 Wistar white rats. The material was obtained from anterior wall of the left ventricle of the heart. To confirm the role of free radicals one group of rats was given ascorbic acid, a well-known substance with anti-oxidation properties (free radical scavenger). In the control group we observed a typical structure of the myocardium. In the group with thyrotoxicosis a great degree of subcellular structural damage was noticed. These alterations correspond to nonspecific myocardial damage that can be meet in hypoxia, reperfusion or in toxic myocardial damage. However, in case of vitamin application generally known to have anti-oxidation properties, damages were significantly less intensive and occurred more rarely. It seems to indicate a significant role of free radicals in the generation of these alterations.

The mast cells and an experimental gastric ulcer. Histochemical, ultrastructural and quantitative study in the rat.

The role of mucosal mast cells (MMC) in chronic non-allergic reactions of the gastric mucosa is unknown. The present study aimed to investigate the reaction of gastric MMC during healing of the acetic acid-induced gastric ulcer in the rat. Mast cells were stained with toluidine blue and alcian blue+safranin and their density and distribution were assessed. Morphometry of MMC and documentation of their contact with eosinophils were based on electron microscopy. In the normal non-injured rat oxyntic mucosa MMC are grouped in two populations: MMC1 concentrated near the glandular necks, and MMC2 in the basal lamina propria. There was a significant decrease in the number of MMC2 near the ulcer, whereas MMC1 lost their concentration in the neck zone. Also the MMC/connective tissue mast cells ratio was increased in the gastric wall adjacent to the ulcer. Eosinophils were commonly in close contact with MMC. Eosinophil cytoplasm adjacent to MMC was devoid of organelles, which were accumulated in the central cytoplasm. The significant redistribution of mast cell population as well as numerous close contacts between mucosal mast cells and eosinophils, taking place in the neighborhood of the chronic gastric ulcer, seem to be not only of morphological but also of functional significance.

Ultrastructure of donor heart muscle prior to transplantation.

Biopsy samples obtained from right ventricle (septum) of donor heart just at the moment of procurement and few hours later after implantation, was examined under electron microscopy. Our study revealed in both biopsy groups the presence of mild cytoplasmic changes similar in character but different in the intensity and extent. The anomalies are unspecific and similar in their character to changes described in disease states or toxic damage. Lack of damage to the membrane system in the cardiac myocytes indicates a reversible character of changes. Further studies are required to compare the present findings with ultrastructural myocardial features after reperfusion.

Apoptosis: genetically programmed physiologic cell loss in normal gastric oxyntic mucosa and in mucosa of grossly healed gastric ulcer.

Maintenance of gastric mucosal structure depends on a dynamic balance between cell loss and cell renewal. The surface epithelial cells exfoliate at a rapid rate (usually in response to luminal contents) and are entirely replaced within 3-5 days. The loss of parietal, chief, and endocrine cells is much slower, but there is little information concerning the morphologic aspects of this process. Because in other tissues cells are physiologically eliminated through a process of apoptosis--genetically programmed cell self-destruction and loss--we studied whether this process occurs in normal gastric mucosa and in the mucosa of recently healed gastric ulcers in the rat. Degeneration and loss of the surface epithelial cells into the gastric lumen occurs in normal oxyntic mucosa, and more extensive desquamation of poorly differentiated mucous cells in the dilated gastric glands takes place within mucosal scars of grossly healed gastric ulcers. Apoptosis of parietal, chief, and endocrine cells in normal oxyntic mucosa and apoptosis of poorly differentiated cells lining dilated glands in mucosal scar were assessed and characterized by electron microscopy. Apoptosis affects single glandular cells and involves a rapid initial condensation of both nucleus and cytoplasm, with subsequent fragmentation. Finally, the cell is converted into a cluster of membrane-bound apoptotic bodies, which usually are engulfed by adjacent cells or disposed into a glandular lumen. Desquamation of surface epithelium and apoptotic self-destruction of glandular epithelium stimulate a constant cell renewal and thus contribute to the maintenance of the ulcer healing process. Rapid and "excessive" proliferation of regenerating gastric epithelium enables re-epithelialization of ulcer crater.(ABSTRACT TRUNCATED AT 250 WORDS)

Gastric injury and invasion of parietal cells by spiral bacteria in rhesus monkeys. Are gastritis and hyperchlorhydria infectious diseases?

The possibility of using the rhesus monkey as a model for studying gastric function in the presence of infection with spiral bacteria was studied. Endoscopic evaluation of the gastric mucosa was performed under general anesthesia in 29 colony-bred rhesus monkeys, and gastric pinch biopsy specimens were obtained from each animal. On a separate day, gastric emptying and acid output were determined using a 99mTc dilution technique. Biopsy samples were fixed for light microscopy (H&E, Gram, and Warthin-Starry stains) and for transmission electron microscopy. The presence of spiral bacteria and gastritis was assessed and rated on coded slides. In 8 of 29 monkeys, Helicobacter pylori-like organisms were observed in close proximity to the mucosal epithelial cells or in the lumen of the gastric pits. In 14 other monkeys, "Gastrospirillum hominis"-like organisms were observed in the mucus covering the surface of epithelial cells, in the lumina of the gastric glands, and overlying parietal cells. Gastritis was present in 8 of 8 animals positive for H. pylori-like organisms, in 2 of 14 animals positive for "G. hominis"-like organisms, and in none of the uninfected monkeys, and the mean gastritis index was significantly greater in animals positive for H. pylori-like organisms. Moreover, acid output was significantly higher in monkeys positive for "G. hominis"-like organisms than in controls or animals positive for H. pylori-like organisms. Gastric emptying was not significantly different in the three groups. In conclusion, (a) H. pylori-like, but not "G. hominis"-like, organisms cause gastritis while not modifying acid output; (b) "G. hominis"-like, but not H. pylori-like organisms, invade and on occasion damage parietal cells while apparently causing hyperchlorhydria; and (c) the rhesus monkey appears to be a good model for the study of gastric infection with spiral bacteria.

Close contacts between mast cells and nerve fibres in the human oxyntic mucosa.

The study of mast cells and nerve fibres in routine biopsies of human oxyntic mucosa was performed with the use of microscopic and ultrastructural methods. Simultaneous visualization of mast cells with Alcian blue and nerves with anti S-100 antibody allowed to study contacts between these two elements of lamina propria. Approximately 17% of mast cells appose nerves in gastric mucosa, which is less than the number of such contacts in the gut reported by other authors. We have found no differences between histologically normal mucosa and gastritis in the aspect of the total number of mast cells per 1 mm2 of lamina propria, number of their contacts with nerves and the ratio of mast cells apposing nerves to the total amount of mast cells. The ultrastructural study revealed significant polymorphism of mast cell-neuron contacts as well as the absence of any specialized structures at the site of adhesion between these two types of cells. The mode of degranulation of mast cells suggests that they are actively engaged in the reaction to noxious stimuli challenging the oxyntic mucosa.