Induction of Multinuclear Cells in the Apical Meristems of Allium cepa by Geomagnetic Field Outrages

This paper is a result of a long-term study of the apical meristems of the Allium cepa L. seedlings against a background of naturally changing geomagnetic field. Multinuclear cells, large cells with large nuclei, and giant cells with giant nuclei were detected. Changes in cellular structures were revealed, and the time-course of this process was followed. Changes in the cellular structure of meristems were correlated with the fluctuations of magnetic field of the Earth. The experiments conducted with in vitro culture of apical meristems showed that the observed changes were regulated on the local level.     

GROWTH AND CELL CYCLE OF TWO CLOSELY RELATED RED TIDE-FORMING DINOFLAGELLATES: GYMNODINIUM NAGASAKIENSE AND G. CF. NAGASAKIENSE1

Small-sized vegetative cells were found to co-occur with normal-sized cells in populations of the European bloom-forming dinoflagellate Gymnodinium cf. nagasakiense Takayama et Adachi, currently known as Gyrodinium aureolum Hulburt, but not in populations of the closely related Japanese species Gymnodiniumn agasakiense. We examined how cell size differentiation may influence growth and cell cycle progression under a 12:12-h light: dark cycle in the European taxon, as compared to the Japanese one. Cell number and volume and chlorophyll red fluorescence in both species varied widely during the photocycle. These variations generally appeared to be related lo the division period, which occurred at night, as indicated by the variations of the fraction of binucleated cells (mitotic index) as well as the distribution of cellular DNA content. “Small” cells of G. cf. nagasakiense divided mainly during the first part of the dark period, although a second minor peak of dividing cells could occur shortly before light onset. In contrast, “large” cells displayed a sharp division peak that occurred 9 h after the beginning of the dark period. The lower degree of synchrony of “small” cells could be a consequence of their faster growth. Alternatively, these data may suggest that cell division is lightly controlled by an endogenous clock in “large” cells and much more loosely controlled in “small” cells. Cells of the Japanese species, which were morphologically similar to “large” cells of the European taxon, displayed an intermediate growth pattern between the two cell types of G. cf. nagasakiense, with a division period that extended to most of the dark period.     

Water movement between epidermal cells of barley leaves – a symplastic connection?

Using the cell-pressure probe the possibility of symplastic water flow between cells of the upper epidermis of barley leaves was investigated. Cells analysed had either an intact or a more or less damaged cellular environment. Cell damage caused large pressure differentials (0·9 MPa) between damaged and adjacent intact cells. Turgor in cells adjacent to damaged cells decreased significantly. Turgor decreases were the larger the more the adjacent, damaged cell was leaking (decreases by 2·5–4·4%). In cells surrounded by a patch of leaking cells, turgor decreased the most, by 18·1–20·4%. In contrast, half-times of water exchange (T_1/2) of cells were not affected by a damaged cellular environment. Assuming that in the barley leaf epidermis, plasmodesmata close at pressure-differentials at or exceeding 0·2 MPa as shown for other plant cells (The Plant Journal 2, 741–750; Canadian Journal of Botany 65, 509–511), it is concluded that symplastic water flow contributes insignificantly to water exchange between cells. Mechanical damage to one individual cell is enough to induce significant turgor changes in neighbouring cells.     

Induction of cell fusion by a factor released by the cellular slime mold Polysphondylium pallidum

Heterokaryons and hybrid cells, which are extremely useful for research in cell biology, can be produced artificially by treating cells with either polyethylene glycol or certain inactivated viruses that alter the plasma membrane. We report here a novel cell-fusion inducing factor secreted by CK-8 strain cells of cellular slime mold Polysphondylium pallidum. Treatment of other strains or other species of cellular slime molds, such as NC-4 of Dictyostelium discoideum with the diluted fraction, containing molecules larger than 50 kDa, of the conditioned medium of CK-8 cell culture induces cell fusion at high frequency and produces multinucleated large cells. This cell fusion is inducible between cells of either a single strain or of two different strains of cellular slime molds.Abbreviations BSS Bonner's salt solution - CM conditioned medium - EDTA ethylenediaminetetraacetic acid - F2 fraction containing cell-fusion induction factor - Mr molecular mass     

Sexual dimorphism among calbindin-D28K immunoreactive cells in the rat pineal body

Calbindin antibodies have been used in neuroanatomical studies to give excellent cytoarchitecural staining and visualization of a Golgi-like cellular morphology. Calbindin-D28K immunoreactivity used in rat pineal gland as a marker detected two classes of pineal cells. One class of small cells representing exclusively glial cells was strongly immunoreactive, and presented a large variety of individual shapes. The majority were a pyramidal shape with one or more processes while others displayed a cytoplasmic lipid droplet. Some small cells occurred around pericapillary spaces. The second class of calbindin-D28K positive cells corresponding to type II pinealocytes were characterized by their large size and less intensive labelling. Type II pinealocytes were round or rectangular; the nucleus was infolded and large with a prominent nucleolus. These large cells were preferentially distributed in the vicinity of vessels and assembled in a cluster of more than ten cells. The lack of S-100 and myeloperoxidase immunoreactivities in large calbindin-D28K cells excluded their possible characterization as glial cells and mononuclear phagocytes, while their size (>15 m) excluded microglial cells. A sex difference was detected between large calbindin-D28K positive cells. The mean calculated number of large positive cells for males was 6361±1504 (n=8) compared to 2162±1235 (n=7) for females. No significative difference was detected between males and females for small calbindin-D28K positive cells.     

Effect of Helicobacter pylori’s vacuolating cytotoxin on the autophagy pathway in gastric epithelial cells

Host cell responses to Helicobacter pylori infection are complex and incompletely understood. Here, we report that autophagy is induced within human-derived gastric epithelial cells (AGS) cells in response to H. pylori infection. These autophagosomes were distinct and different from the large vacuoles induced during H. pylori infection. Autophagosomes were detected by transmission electron microscopy, conversion of LC3-I to LC3-II, GFP-LC3 recruitment to autophagosomes, and depended on Atg5 and Atg12. The induction of autophagy depended on the vacuolating cytotoxin (VacA) and, moreover, VacA was sufficient to induce autophagosome formation. The channel forming activity of VacA was necessary for inducing autophagy. Intracellular VacA partially co-localized with GFP-LC3, indicating that the toxin associates with autophagosomes. The inhibition of autophagy increased the stability of intracellular VacA, which in turn resulted in enhanced toxin-mediated cellular vacuolation. These findings suggest that the induction of autophagy by VacA may represent a host mechanism to limit toxin-induced cellular damage.     

Ribosomal DNA clusters in pulsed-field gel electrophoretic analysis of human acrocentric chromosomes

For determination of the extent to which ribosomal DNA (rDNA0 is organized in tandemly repeated arrays, cellular DNA was digested with a restriction enzyme (EcoRV) that does not cut within the single 44-kb rDNA unit, and fragments separated by PFGE were hybridized to specific rDNA probes. A series of bands large enough to contain 15 to more than 30 rDNA repeat units was observed. In YACs containing cloned rDNA, however, such clusters were not observed, presumably because, as shown here for a clone starting with 1.5 tandem repeat units, there is a tendency for repeat units to delete out of the insert. By comparative gel electrophoretic analyses of DNAs from rodent hybrid cells containing singly isolated human chromosomes, most of the bands seen in total human DNA were assigned to at least one of the acrocentric chromosomes. Thus, large characteristic assemblies of DNA containing rDNA and lacking EcoRV sites were stable enough to be conserved in some human/rodent hybrid lines. When further digested with HindIII, which cuts rDNA at several points, the rDNA in each band yielded the expected fragments. If the large species consist completely of clusters of tandemly repeated rDNA units, they account for about half of the total cellular rDNA content estimated by saturation hybridization measurements.     

ULTRASTRUCTURE OF MAIZE ENDOSPERM SUSPENSION CULTURES

Suspension cultures derived from developing maize (Zea mays L.) endosperm were examined by electron microscopy, after both glutaraldehyde-OsO₄ and KMnO₄ fixation, and compared with intact endosperm. Tissue clumps consisted of interconnected cell clusters without any organization of the different cell types. The cultures were comprised of cells with dense cytoplasm and small vacuoles, large vacuolate cells, and cells in which storage products (starch, protein bodies, or lipid) accumulated. The endomembrane system of cultured cells was more highly developed than that of cells of the intact endosperm. In particular, arrays of smooth endoplasmic reticulum were seen only in the cultured cells. An abundance of endoplasmic reticulum, dictyosomes, and ribosomes is consistent with the recently reported extracellular secretion of enzymes by these cultures. Cell wall ingrowths, a characteristic of basal endosperm transfer cells, were observed occasionally in cultured cells, but cells with ingrowths had no histological organization. Some of the observed features may have resulted from perturbation of normal cellular events caused by the conditions of in vitro growth. These cultures are a useful tool for studying cellular mechanisms of protein secretion and storage product accumulation in developing maize endosperm.     

Understanding Saline and Osmotic Tolerance of Populus euphratica Suspended Cells

Tolerance of Populus euphratica suspended cells to ionic and osmotic stresses implemented respectively by NaCl and PEG (6000) was characterized by monitoring cell growth, morphological features, ion compartmentation and polypeptide patterns. The cells grew and proliferated when submitted to stresses of 137 mM NaCl or 250 g l⁻¹ PEG, and survived at 308 mM of NaCl, showing tolerance to saline and particularly osmotic stress. They were resistant to plasmolysis and had dense cytoplasms, large nuclei and nucleoli, and evident cytoplasmic strands under high saline and osmotic stress. The sequestration of Cl⁻ into the vacuoles was observed in the cells stressed with 137 and 223 mM NaCl. The cellular protein profile was modified by high salt and osmotic stress and showed 28 kDa polypeptides up-regulated by both NaCl and PEG, and 66 and 25 kDa polypeptides up-regulated only by high NaCl stress. The salt tolerance of P. euphratica cells might be related to their capacity of adapting to higher osmotic stress by maintaining cell integrity, sequestrating Cl⁻ into vacuoles and modulating polypeptides that reflect cellular metabolic adaptations.     

Changes in <Emphasis Type="Italic">Synechococcus</Emphasis> Population Size and Cellular Ribosomal RNA Content in Response to Predation and Nutrient Limitation

A mathematical model of predator-prey interactions was used to predict the relationship between population size and cellular growth rate in a two-tiered trophic system consisting of Synechococcus PCC 6301 and Tetrahymena pyriformis. As predicted, axenic chemostat cultures of Synechococcus responded to increased nutrient availability by expanding the equilibrium population size without a concurrent change in growth rate. Likewise, the addition of the predator Tetrahymena pyriformis decreased the Synechococcus population size by 85% and increased the Synechococcus growth rate. Synechococcus populations in the surface waters of the Gulf of Mexico were sampled to ascertain whether the relationship between population size and cellular 16S rRNA concentration conformed to that predicted by the model. Direct counts of autofluorescent cells in size-fractionated seawater samples provided an estimate of Synechococcus population size. The growth rate of in situ populations was estimated by measuring the extent of hybridization of an oligonucleotide probes complementary to Synechococcus 16S rRNA, based on evidence that ribosomal RNA content increases concurrently with growth rate. The comparison of in situ population sizes and specific growth rates revealed that relatively large Synechococcus populations were growing slowly, indicative of nutrient limitation, and that quickly growing populations were relatively small, as predicted for predator-limited populations.     

Immunocytochemical localization of cathepsins B,H, and L in the rat gastro-duodenal mucosa

Summary Cathepsins B, H, and L are representative cysteine proteinases in lysosomes of a large variety of cells. Previous immunochemical studies indicated the presence of these enzymes also in the gastrointestinal wall. Using specific antisera, the cellular and subcellular distribution of cathepsins B, H, and L in rat gastric (oxyntic and pyloric part) and duodenal mucosa was investigated by light and electron microscopical immunocytochemistry. The subtypes of cathepsins were distributed differently in the cellular constituents of the epithelia: Cathepsin B was localized to lysosomes of all cells except goblet cells. Cathepsin H was found predominantly in gastric parietal cells (lysosomes) and in secretion granules of pyloric gastrin and duodenal cholecystokinin cells. Cathepsin L immunoreactivities were weak and restricted to a minority of cells (gastric mucous cells, enterocytes). Interstitial cells of the lamina propria immunoreactive for cathepsins H and L were identified as macrophages. The present findings suggest a dual function of cathepsins in the gastro-duodenal mucosa. They (1) cleave enzymatically proteins and peptides ingested in lysosomes, and (2) they may be involved in the processing of biologically active peptides (enteric hormones) from their precursor proteins.     

The loss of contact inhibition and anchorage-dependent growth are key steps in the acquisition of Listeria monocytogenes susceptibility phenotype by non-phagocytic cells

Summary— We have previously demonstrated that intestinal and kidney finite cell lines were resistant to L monocytogenes invasion (ie allowed low bacterial entry and no intracellular multiplication) in contrast to the continuous cell lines which were susceptible to Listeria invasion (ie allowed high bacterial entry and intracellular multiplication) (Velge et al (1994a) Med Microbial Immunol 183, 145). The aim of this study was to discover whether epigenetic or genetic cellular modifications could convert L monocytogenes resistant cells into a susceptible phenotype and to determine the cellular steps involved in Listeria susceptibility. Among the 5-azacytidine treated finite cell lines, the untransformed immortal cell lines established remained resistant to L monocytogenes invasion whereas the weakly transformed continuous cell lines established were converted into a susceptible phenotype. Transfection of resistant cells by SV40 large T antigen induced only highly transformed continuous cell lines displaying a susceptible phenotype. Taken together these data show that cell transformation enhanced Listeria invasion. This conclusion was supported by the observation that L monocytogenes was able to induce cell foci within murine finite cell monolayers. This morphological cell transformation was completely reversible and required live bacteria inside cells. In conclusion, we may speculate that the L monocytogenes intracellular multiplication observed within cell foci could be explained by the loss of contact inhibition of the finite cell monolayer. Indeed, the loss of both contact inhibition and anchorage-dependent growth are the key steps involved in the L monocytogenes susceptibility phenotype.     

Ultrastructural aspects of kranz anatomy inDigitaria sanguinalis andSetaria viridis (poaceae)

Structural differentiation of Kranz anatomy has been investigated in leaf cross sections of two C-4 Poaceae:Digitaria sanguinalis andSetaria viridis. The study mainly focused on cellular and interfacial features of bundle sheath (BS) and mesophyll (MS) cells of the C-4 structure. Prominent BS, spaced by only two MS cells apart, were surrounded concentrically by a layer of MS cells. BS cells ofS. viridis had centrifugally arranged relatively large chloroplasts containing much starch, but the chloroplasts had agrana to rudimentary grana. Structural and size dimorphisms, when starch was present, were detected between BS and MS chloroplasts. Loosely arranged MS cells had peripherally displaced smaller chloroplasts containing little to none starch. BS chloroplasts ofD. sanguinalis were similar to those ofS. viridis, but had very little starch and well-developed long agranal stroma lamella. Features of MS cells were similar in both species, but well-defined peripheral reticulum (PR) was easily recognized in MS chloroplasts ofS. viridis. Virtually no PR was developed in BS chloroplasts examined. BS cells contained more mitochondria and microbodies, but no structural dimorphism was noticed. The electron-dense suberized lamella were often observed between BS and MS cells, especially in the primary wall of BS cells. It was most frequently found at the BS and MS cell interfaces and terminated in radial walls of the adjacent BS cells. Prominent pits with plasmodesmata (pd) were seen in the walls of both cells. There also were numerous pd in outer tangential walls of the BS cells. The number of pd ranged from 20 to 60. The pd trasversed a segment of cell wall much thinner than the adjacent wall. The current cellular data have been compared to the ultrastructural features known in leaves of other C-4 plants, especially NADP-ME species.     

Cyclic AMP and calcium exchange in a cellular slime mold

Cyclic AMP is known to be an effective chemotactic agent for amebae of the cellular slime mold D. discoideum. A large amount of information from experiments on metazoa suggested that one cellular effect of cyclic AMP might be to alter the permeability of the cell membrane to Calcium or Sodium ions. On the basis of this information experiments were designed to test the effect of cyclic AMP on outflow of labeled Calcium or Sodium ions from amebae of D. discoideum. It was found that addition of cyclic AMP at 10^?4M resulted in a large increase of Ca⁴⁵ outflow from cells at the pre-aggregative or aggregative stage of development. No effect was found on Na²² outflow. It is suggested that this effect on Calcium permeability of the membrane is related to the chemotactic influence of ATP by some action on the contractile mechanism for ameboid movement. The phenomenon may be distinct from the enzyme inductive activity of cyclic AMP known for bacteria, and perhaps occurring in the cellular slime molds as well.     

Glial cells phagocytose neuronal debris during the metamorphosis of the central nervous system in Drosophila melanogaster

 Using electron microscopy we demonstrate that degenerating neurons and cellular debris resulting from neuronal reorganization are phagocytosed by glial cells in the brain and nerve cord of the fruitfly Drosophila melanogaster during the first few hours following pupariation. At this stage several classes of glial cells appear to be engaged in intense phagocytosis. In the cell body rind, neuronal cell bodies are engulfed and phagocytosed by the same glial cells that enwrap healthy neurons in this region. In the neuropil, cellular debris in tracts and synaptic centres resulting from metamorphic re-differentiation of larval neurons is phagocytosed by neuropil-associated glial cells. Phagocytic glial cells are hypertrophied, produce large amounts of lysosome-like bodies and contain a large number of mitochondria, condensed chromatin bodies, membranes and other remains from neuronal degeneration in phagosomes. Received: 23 January 1996 / Accepted in revised form: 21 May 1996     

Physiological and Optical Responses of the Harmful Dinoflagellate Heterocapsa circularisquama to a Range of Salinity

The responses of division rate, cell volume, cellular carbon (C) and nitrogen (N), cellular pigments and optical characteristics to changes in salinity were examined in the dinoflagellate Heterocapsa circularisquama. The physiological and optical characteristics of H. circularisquama were significantly affected by changes in salinity. When cells were exposed to different salinities, they exhibited physiological acclimation by adjusting their cellular properties associated with growth. This could be a beneficial tactic for ensuring proliferation and limiting damages induced by adverse environmental factors. The results of this study could be essential for assisting in the development of growth models for H. circularisquama.     

Two-dimensional gel electrophoretic analysis of Escherichia coli proteins: influence of various anaerobic growth conditions and the fnr gene product on cellular protein composition

Two-dimensional gel electrophoresis was used to examine the response of the cellular proteins of Escherichia coli to various anaerobic growth conditions and to the presence or absence of a functional Fnr protein. The steady-state levels of 125 polypeptides were found to vary in either a positive or negative manner, with many polypeptides being affected under a number of conditions. A large number (21) of the anaerobically inducible polypeptides were shown to be totally independent of the presence of Fnr while 22 were shown to be reduced in a fnr mutant under all anaerobic growth conditions tested. A total of 8 proteins were shown to be reduced in a fnr mutant only in aerobically grown cells indicating that the Fnr protein has a function in the presence of oxygen. This was further confirmed by the observation that 15 anaerobically inducible polypeptides were also found to show an increase in aerobically grown cells, however, only in a fnr strain. This latter finding implies that Fnr may also exhibit repressor function. This effect of Fnr-dependent repression was also observed with several polypeptides in anaerobically grown cells.Abbreviation CRP cyclic AMP receptor protein     

Anti-infective control in human bronchiolar epithelial cells by mucin phenotypic changes following uptake of N-acetyl-L-cysteine

Purpose. Aspiration pneumonia is infection of the respiratory tract resulting from accumulation of sputum in the larynx. N-acetyl-L-cysteine (NAC) might regulate mucin (MUC) expression and activate inherent anti-infective system in bronchiolar epithelial cells after cellular uptake, and therefore, serve as the preventative agent for chronic lung disease including aspiration pneumonia. The purpose of this in vitro study was to evaluate the effect of uptake of NAC by human bronchiolar epithelial cells on bacterial infection and regulations of mucin expression in association with cellular redox status under co-culture with a representative pathogen for hospital- and community-acquired pneumonia, Streptococcus pneumoniae. Materials and methods. Human bronchiolar epithelial cells preincubated with or without 20 mM NAC for 3 h were co-cultured with or without bacteria for 8 h and evaluated with respect to cellular redox balance, expressions of various types of MUC, proinflammatory cytokines and mediators, and bacterial infection state by biochemical, genetic, and immunofluorescent assays. Results. Markedly increased intracellular reactive oxygen species and oxidized glutathione levels plus increased release and expression of proinflammatory cytokines and mediators were observed in cells co-cultured with bacteria. These bacteria-induced cellular redox disturbance and proinflammatory events were prevented and alleviated by pretreatment with NAC. Cells co-cultured with bacteria did not increase expression of anti-infective membranous MUC4 but exhibited increases in gel-forming MUC5AC expression and bacterial infection. However, NAC-pretreated cells avoided bacterial infection along with enhancement of MUC4, but not MUC5AC, expression. Conclusion. Uptake of NAC by human bronchiolar epithelial cells prevented bacterial infection and upregulated membranous, but not gel-forming, MUC expression along with the increase in intracellular antioxidant level under co-culture conditions with S. pneumoniae.     

Effects of 47C allele (rs4880) of the SOD2 gene in the production of intracellular reactive species in peripheral blood mononuclear cells with and without lipopolysaccharides induction

Abstract

Challenging of peripheral blood mononuclear cells (PBMCs) with lipopolysaccharides (LPS) has been shown to activate monocytes and macrophages, leading to the production of pro-inflammatory cytokines and reactive oxygen species (ROS). Manganese superoxide dismutase (MnSOD) is an important enzyme that may play a central role in the response to oxidative stress. 47C> T SNP of the SOD2 gene, the -9Val MnSOD is less efficient than the -9Ala version. We have previously characterized the cellular redox status of human PBMCs expressing either -9Ala (CC) or -9Val (TT) SOD2 and analyzed the responses of these cells to oxidative stress induced by LPS. Due to the observed alterations in the activities of these antioxidant enzymes, we decided to investigate their immunocontent and analyze the production of intracellular oxidants, as well as any resulting DNA damage. PBMCs were isolated from the blood of 30 healthy human volunteers (15 volunteers per allele). We then analyzed levels of nitrite, DNA damage by comet assay, TNF-α, carboxymethyl lysine and nitrotyrosine and assessed production of intracellular reactive species by the DCFH-DA-based assay and western blots were used to analyze protein levels. Our results show that there occurs an increase in nitric oxide production in both allele groups after challenge with LPS. A significant increase in DNA damage was observed in PBMCs after an 8-h LPS challenge. Cells expressing the SOD2 47C allele quickly adapt to a more intense metabolism by upregulating cellular detoxification mechanisms. However, when these cells are stressed over a long period, they accumulate a large quantity of toxic metabolic byproducts.