Antagonistic effect of bacteriocinogenic Lactobacillus acidophilus on Klebsiella pneumoniae, Citrobacter freundii and Proteus mirabilis cells

Data of the ultrastructural cellular changes of conditionally pathogenic enterobacteria, including K. pneumoniae, C. freundii and P. mirabilis cells impacts to bacteriocin-producing L. acidophilus are presented. Enterobacteria in response to the bacteriocinogenic effect of lactobacilli are manifestated by expressive destructions of sensitive to pore formation bacteriocin cells. Various morphological types of enterobacteria cells with increase of involution, lysing and resting forms are revealed. The specific ultrastructural changes of enterobacteria cells which evidencing the significant destructive processes of the cells membranes are detected. The destabilization of cellular wall in expansion periplasmic spaces and appearance of the ultrastructural reorganization of bacterial cells nucleoid also are registrated. Revealing the mechanism of lactobacilli secreted bacteriocin action to conditionally pathogenic enterobacteria might provide new ways to select the effective highly antagonistic probiotic strains.     

Ultrastructure of epithelial cells in rat's epididymal caput in experimental hyperprolactinemia induced by metoclopramide

The aim of the performed studies has been to find out wether or not the ultrastructural alterations of the epithelial cells in the rat's epididymal caput occur in hyperprolactinemia induced by metoclopramide and to see if the observed changes are of reversible character. It has been revealed that prolactin concentration was twice as high as in control animals due to peritoneal administration of metoclopramide in a dose of 2.2 mg/kg body mass, given for 14 days. The ultrastructural alterations in the principal cells of epididymal caput affected cellular organelles being involved in proteins synthesis, glycosylation, secretion as well as energetic processes. They were manifested by decreased amount of rough endoplasmic reticulum, widening of its cisternae combined with degranulation, distension of Golgi apparatus cisternae, elevated number of vesicles in apical part of the cells, and changes in mitochondria. The termination of metoclopramide administration made prolactin concentration exhibit values being almost similar to those determined in control rats, whereas the ultrastructural changes in the principal cells were found to be reversible.     

Ultrastructural characteristics of the incretory granules of the EC- and ECL-cells of the gastrointestinal tract

At least three varieties of EC-cells differing in their density, form and fine structure can be revealed at the ultrastructural study of EC- and ECL-cells of the intestinal tract. Different forms of the incretory granules of the ECL-cells and heterogenous ultrastructural organization of their dense contents in man and animals make is possible not only to distinguish several forms of these cellular elements, but to speak of their specific peculiarities. It is not impossible that the ultrastructural peculiarities revealed in the incretory granules of the endocrine cells explain differences in the nature of histologically active substance which they produce.     

Intracellular and extracellular activity of cathepsin D in the liver in cirrhosis and its involution

The distribution of cathepsin D in liver with CCl4 induced cirrhosis and its involution in rats was investigated by ultrastructural cytochemistry. Besides intracellular, it was revealed the extracellular activity of cathepsin D. The reaction product was on collagen fibers near the hepatocytes and connective tissue cells as well as on the hepatocytes microvilli and on the outside part of cellular membrane of connective tissue cells (macrophage, fibroblast, Ito cells). Hence the source of extracellular cathepsin D in liver are the parenchymatous as well as nonparenchymal cell elements. The results testify that under the cirrhosis and its involution, the cathepsin D takes part in intracellular proteolysis and is secreted by hepatocytes and connective tissue cells in the intracellular space; it also takes part in extracellular catabolism of connective tissue.     

The electron microscopic study of cell-to-cell interactions between antagonistic microorganisms

The electron microscopic study of thin sections and positively stained specimens of cells taken from particular cocultures of Lactobacillus acidophilus D75, Lactobacillus casei YIT 9018, Shigella flexnery 2a, Bacillus subtilis ATCC 6633, and Staphylococcus aureus ATCC 25923 (some of these bacteria are antagonistic to others) showed the presence of specific ultrastructural elements indicating cell specialization and cooperation. The responses of antagonistic bacteria manifested themselves at the cellular and population levels.     

Electron microscopy and autoradiography study of bronchoalveolar lavage cells in nonspecific pneumonia

Cells obtained by bronchoalveolar lavage from patients with chronic nonspecific pulmonary inflammatory diseases were studied using light and electron microscopy and radioautography. Five morphological forms of alveolar macrophages, distinct in their structure and 3H-uridine content were described. A higher level of RNA synthesis has been revealed in alveolar macrophage forms 2 and 3 than in forms 1, 4 and 5; with it being lower in polymorphonuclear leukocytes and lymphocytes. It was shown that changes in the number of lavage cells and the structure-to-function characteristics in each cellular population depended on the phase of the inflammatory process. It was postulated that structural and metabolic heterogeneity of alveolar macrophages reflected the successive stages of cellular development from cell-precursors (through activation of protein synthesis) to cells with complete lysosomal cycle and the following phagocytosis.     

The ultrastructural changes in the glomeruli and juxtaglomerular apparatus at different periods of endotoxic shock]

It is shown that after endotoxin injection the ultrastructural changes in the glomeruli can favour development of the acute renal insufficiency. In the initial and intermediate periods of the endotoxin shock the granular and agranular forms of juxtaglomerular cells hyperfunction, respectively, are revealed as well as an increase of renin activity in plasma. At the stage of the late endotoxemia ultrastructural alterations are stabilized. The juxtaglomerular cells synthesize and accumulate secretory granules, but renin activity in plasma decreases almost to the initial level.     

Effects of wheat germ agglutinin on the cellular content of filamentous actin in Intestine 407 cells

Increasing experimental evidence suggests that gluten contains a toxic factor that may cause ultrastructural changes in the small intestine which mimic those found in patients with celiac disease. In addition, it has recently been proposed that the toxic factor of gluten is a protein very similar, if not identical, to a well known lectin, wheat germ agglutinin (WGA). Since the cytoskeleton forms the basis of the ultrastructural architecture of the enterocytes the present study was performed to investigate whether WGA has a direct effect on the cytoskeleton in Intestine 407 cells. Changes in the cellular content of filamentous actin (F-actin) in these cells were studied with the fluorescein isothiocyanate (FITC)-phallacidin assay. Cellular exposure to WGA led to a rapid reduction in the cellular content of F-actin (greater than 50%). Intracellular buffering of the cytosolic free calcium level using quin2 as a chelator of calcium totally abolished the WGA-induced reduction in F-actin content. However, increasing the cytosolic free calcium level by exposure to the calcium ionophore ionomycin did not affect the cellular content of F-actin. Ionomycin also failed to potentiate the effect of WGA on the cellular F-actin content. The present results show that WGA changes the organization of the cytoskeleton in Intestine 407 cells via a calcium-dependent mechanism, however, in addition to calcium, some other signal(s), possibly an increased turnover of the phosphatidylinositol cycle, is(are) also required.     

Ultrastructural changes in the thymus of the turtle Mauremys caspica in relation to the seasonal cycle

Summary Changes in the ultrastructure of the thymus of the turtle Mauremys caspica, with special reference to its non-lymphoid components, were studied in relation to the seasonal cycle. The thymic cortex contains framework-forming epithelial-reticular cells and free macrophages, while the medulla includes, in addition, mature and presumptive pro-interdigitating cells. The ultrastructural features of these cells are generally similar to those described for non-lymphoid components of the mammalian thymus. The turtle thymus undergoes cortical involution in spring, with recovery periods in May–June and during autumn. A moderate involution occurs in winter. At the beginning of spring, cortical (but not medullary) epithelial-reticular cells show degenerative changes, probably related to high levels of circulating testosterone. In spring and autumn, mature interdigitating cells are absent, but macrophages, monocytes, and pro-interdigitating cells are found. During May–June, the cortical epithelial-reticular population recovers and macrophages, monocytes, and interdigitating cells are actively phagocytic. In summer, the epithelial-reticular cells in both cortex and medulla display normal ultrastructural features; mature and immature interdigitating cells are absent and some macrophages are detected occasionally. The results suggest that non-lymphoid components of the reptilian thymus can play a role in governing T-lymphocyte differentiation, and that the thymic cortex and medulla exhibit different cycles of seasonal activity.     

The electron microscopic study of cell-to-cell interactions between antagonistic microorganisms

The electron microscopic study of thin sections and positively stained specimens of cells taken from particular cocultures of Lactobacillus acidophilus D75, Lactobacillus casei YIT 9018, Shigella flexnery 2a, Bacillus subtilis ATCC 6633, and Staphylococcus aureus ATCC 25923 (some of these bacteria are antagonistic to others) showed the presence of specific ultrastructural elements indicating cell specialization and cooperation. The responses of antagonistic bacteria manifested themselves at the cellular and population levels.     

Increased activity of plasminogen activators during involution of the rat ventral prostate.

Plasminogen activator was measured in the ventral prostates of non-castrated, castrated, and androgen-treated rats to determine whether changes in this activity correlated with the process of glandular involution. While the activity was very low in cytosolic extracts from the prostates of non-castrated rats, 2 days following castration the plasminogen activator activity increased in a near-linear fashion such that by day 7 it was 10-fold higher in terms of specific activity (per mg of protein) and cellular concentration (per mg of DNA). During this interval there was a rapid decrease in the cell population of the prostates. Treatment of the 7-day castrated rats with the potent androgen, dihydrotestosterone, both reduced the plasminogen activator activity and restored the cell number in a dose-related manner. Gel electrophoretic analysis revealed two major bands of plasminogen activator activity in the cytosolic extracts from 4- and 7-day castrated rats, plus additional minor bands in samples from 10- and 14-day castrated rats. Approx. 10% of the cellular concentration of plasminogen activator activity was recovered in association with an 18000g pellet fraction from the prostates; this fraction showed less heterogeneity of the plasminogen activator forms as observed by gel electrophoresis. Inhibitor studies indicated that the 18000g pellet fraction from the prostates of non-castrated rats possessed some plasminogen activator inhibitor activity, but the relative concentration of the inhibitor activity was small. We conclude that the involution of the prostate is probably associated with increased synthesis of plasminogen activators through a de-repression process which may involve loss of androgen receptors.     

Morphological changes of in-vitro-produced bovine blastocysts after vitrification,in-straw direct rehydration,and culture

Morphological signs of injury and regeneration following vitrification and warming of bovine embryos were studied by light and electron microscopy. In-vitro-produced Day 7 expanded blastocysts (Day 0 = day of insemination) were vitrified by a two-step equilibration method using ethylene glycol and dimethyl sulphoxide as cryoprotectants. Thawing was performed by in-straw direct rehydration, followed by in vitro culture on a granulosa cell monolayer. Embryos were processed for transmission electron microscopy immediately after warming (0 hr) as well as after 4 hr or 24 hr of culture following warming. A control group of unfrozen embryos was also processed. At 0 hr after warming, except for a rapid collapse of the blastocoele, only minor changes were detectable by stereomicroscope. However, at the ultrastructural level, signs of extensive injury were seen, including a general distension or shrinkage of mitochondria, disintegration of cell adhesions between adjacent trophoblastic cells, and complete rupture of some cells. At 4 hr, stereomicroscopic investigation revealed collapsed blastocoele and a darkened granular appearance of the cell mass. At the ultrastructural level, signs of regeneration were also observable: cells with minor injuries were re-assembled in a central area forming a small blastocoele, cell adhesion structures were re-established, and damage of mitochondria was less severe. The majority of irreversibly damaged cells or cell debris was accumulated in the perivitelline space. At 24 hr, stereomicroscopic investigation of surviving blastocysts showed no signs of the previous injury. At the ultrastructural level, cellular debris in the perivitelline space and some degenerated cells in the blastocoele were the only signs of previous injuries. In conclusion, ultrastructural investigation revealed unexpectedly extensive damage followed by a rapid regeneration and reorganization of the embryonic structure. Mol. Reprod. Dev. 48:9–17, 1997. © 1997 Wiley-Liss, Inc.     

Atypical cell forms overproducing extracellular substances in population of cycad cyanobionts

The ultrastructure of the cyanobionts of the greenhouse-grown cycads Cycas circinalis, Ceratozamia mexicana, and Encephalartos villosus was studied. The cyanobiont microcolonies grown in the intercellular space of the cyanobacterial zone of cortical parenchyma in the cycad coralloid roots contained two specific forms of vegetative cells with a reduced cell wall, namely, protoplasts and spheroplasts. The protoplasts and spheroplasts exhibited ultrastructural changes indicating the overproduction of two extracellular substances, one of which resembled the mucilage polysaccharides and the other was proteinous. The substances were likely to be synthesized intracellularly and then be excreted with the aid of surface vesicles or by channels in the cytoplasmic membrane to form, respectively, a slimy extracellular matrix and an additional electron-opaque envelope around the cell. At the late developmental stages, the excretion of these substances was accompanied by degradative changes in the cells, leading eventually to cell death. The physiological role of these specific cell forms and the factors that induce their development and death in the cell populations of cyanobionts are discussed.     

Fine structure of the parathyroid glands in baboons, Papio hamadryas in response to experimental hypercorticoidism

Female baboons maintained under laboratory conditions were subjected to a series of 10 weekly injections (4 mg/kg body weight) of the synthetic glucocorticoid, triamcinolone hexacetonide. In response to the treatment, serum immunoreactive parathyroid hormone (PTH) levels were raised, though blood calcium levels remained within normal physiological limits. Light and electron-microscopic studies were made on the parathyroid glands at the end of the experimental period. The baboon parathyroid glands were composed of 'light' and 'dark' forms of the chief cells in varying ratios from gland to gland even within a single animal. Glucocorticoid-induced parathyroid hyperactivity as measured by circulating PTH levels was not accompanied by cellular hypertrophy, though there was an increase in the relative number of 'light' cells. At the ultrastructural level, after treatment, many of the 'light' cells were found to contain more free ribosomes, larger profiles of granular endoplasmic reticulum and had better developed mitochondria. Interdigitations between adjacent chief cells were more complex in treated glands. Apart from these features, chief cells of treated glands were basically similar to those of untreated controls. Our study showed that functional parathyroid hyperactivity in baboons is not necessarily accompanied by significant ultrastructural changes in chief cells.     

Morphofunctional features of the secretory cells of the adenohypophysis and adrenal glands of the rat during aging

Karyometric and electron microscopic investigation has been performed in the adenohypophyseal secretory cells, in the cortex and medulla of the adrenal glands in mature (6 months) and ageing (25-26 months) white male rats. The diameter of the corticotropic cell nuclei of the adenohypophysis significantly increases with age. In the secretory cells of the adrenal cortex and medulla no important changes in their parameters are revealed. With age destructive changes of the ultrastructures are manifested variously in different types of the secretory cells in the endocrinic glands studied. In the corticotropic, somatotropic, thyreotropic cells of the adenohypophysis, in spongiocytes of the adrenal cortex, compensatory ultrastructural rearrangements develop; they are aimed to preserve functional activity of the cells mentioned. No age changes are revealed in ultrastructure of chromaffin cells of the adrenal medulla.     

Gonadotrophs following removal of the ovaries: a fine structural study of human pituitary glands.

The fine structure of gonadotrophs has been investigated in surgically removed pituitary glands of 12 women who because of disseminated breast cancer, underwent bilateral ovariectomy at various periods before hypophysectomy. Compared with the adenohypophyses of 3 non-ovariectomized female subjects with diabetes mellitus, electron microscopy revealed that two cell types were affected by gonadectomy. These cell types corresponded to those which were regarded as FSH gonadotrophs and LH gonadotrophs in previous studies. In addition in the adenohypophyses stimulated by removal of the ovaries, intermediary cell types began to appear suggesting a transformation of LH gonadotrophs to FSH gonadotrophs. The most conspicuous change following gonadectomy was the formation of castration cells. These cells arose from FSH gonadotrophs and exhibited ultrastructural features interpreted as representing the morphologic manifestations of sustained hypersecretion of gonadotrophins. It seemed that castration cells have a limited life span and in their advanced stages of development they show ultrastructural signs indicative of irreversible involution.     

IL-7 gene therapy in aging restores early thymopoiesis without reversing involution

Thymic involution begins early in life and continues throughout adulthood, resulting in a decreased population of naive T cells in the periphery and a reduced ability to fight off newly encountered infectious diseases. We have previously shown that the first step of thymopoiesis is specifically blocked in aging. This block at the DN1 to DN2 transition and the subsequent loss of thymic output in old age mirrors the changes seen in IL-7-deficient mice, and it is hypothesized that decreased intrathymic IL-7 is involved in age-related thymic involution. To separate the effect of IL-7 on thymic involution from its function as a peripheral lymphocyte growth cofactor, we injected IL-7-secreting stromal cells into the thymi of recipient mice. The increased local concentration of IL-7 maintained the first step of thymopoiesis at a level far higher than was seen in age-matched controls. However, despite this success, there was no decrease in thymic involution or increase in T cell output. The inability of IL-7 to prevent involution led us to the discovery of an additional age-sensitive step in thymopoiesis, proliferation of the DN4 population, which is unaffected by IL-7 expression.     

Truncation of the beta-catenin binding domain of E-cadherin precedes epithelial apoptosis during prostate and mammary involution

A potential target of hormone action during prostate and mammary involution is the intercellular junction of adjacent secretory epithelium. This is supported by the long-standing observation that one of the first visible stages of prostate and mammary involution is the disruption of interepithelial adhesion prior to the onset of apoptosis. In a previous study addressing this aspect of involution, we acquired compelling evidence indicating that the disruption of E-cadherin-dependent adhesion initiates apoptotic programs during prostate and mammary involution. In cultured prostate and mammary epithelial cells, inhibition of E-cadherin-dependent aggregation resulted in cell death following apoptotic stimuli. Loss of cell-cell adhesion in the nonaggregated population appeared to result from the rapid truncation within the cytosolic domain of the mature, 120-kDa species of E-cadherin (E-cad(120)). Immunoprecipitations from cell culture and involuting mammary gland demonstrated that this truncation removed the beta-catenin binding domain from the cytoplasmic tail of E-cadherin, resulting in a non-beta-catenin binding, membrane-bound 97-kDa species (E-cad(97)) and a free cytoplasmic 35-kDa form (E-cad(35)) that is bound to beta-catenin. Examination of E-cadherin expression and cellular distribution during prostate and mammary involution revealed a dramatic reduction in junctional membrane staining that correlated with a similar reduction in E-cad(120) and accumulation of E-cad(97) and E-cad(35). The observation that E-cadherin was truncated during involution suggested that hormone depletion activated the same apoptotic pathway in vivo as observed in vitro. Based on these findings, we hypothesize that truncation of E-cadherin results in the loss of beta-catenin binding and cellular dissociation that may signal epithelial apoptosis during prostate and mammary involution. Thus, E-cadherin may be central to homeostatic regulation in these tissues by coordinating adhesion-dependent survival and dissociation-induced apoptosis.     

Dynamics of acid phosphatase activity in the liver in the process of involution of cirrhosis

Changes in the total activity of acid phosphatase in the liver as well as changes in the enzyme activity in hepatocytes and connective tissue cells of fibrosis layers were investigated, using quantitative histochemical method, in the process of mouse cirrhosis involution. After discontinuation of CCl4 injection, the animals with cirrhosis were divided into two groups. In the first group the resection of the left lobe of the liver was performed. The animals of the second group were not subject to operation. The results demonstrate that there is a close correlation between lysosomal hydrolase activity of hepatocytes and connective tissue cells of the liver and collagen resorption during cirrhosis involution. The most intensive lysis of collagen takes place within the first three weeks of cirrhosis involution in both experimental groups. Partial resection in cirrhosis has no significant effect on the changes and level of total activity of lysosomal hydrolase enzymes in the liver during cirrhosis involution.     

Disorganization of mitosis in HeLa cells by deuterium oxide

We assessed, by light and electron microscopy, the influence of deuterium oxide on the dynamics of mitosis and on the morphology of the mitotic apparatus in HeLa cells grown in vitro. A 2-h incubation of HeLa monolayers with low concentrations of D2O (1%-25%) in the medium increased the frequency of multipolar divisions up to 20 times the control level. Substitution of 10% and 25% D2O for H2O induced changes in the proportions of mitotic phases. These changes could be fully reversed to the control pattern after 1 h of recovery in non-deuterated medium. Fifty % D2O strongly inhibited, and 75% D2O blocked the cell cycle before prophase and at (pro-)metaphase. In cells treated with 50% D2O conspicuous morphological changes of the interphase chromatin as well as ultrastructural abnormalities of all mitotic phases were regularly observed. Overall, these results confirmed the antimitotic activity of deuterium oxide and revealed that it could also influence the cell cycle before mitosis. It is suggested that interference with diverse cellular constituents rather than a specific influence on microtubule turnover could be responsible for the disorganization of the cell cycle in HeLa cells by D2O.