The three-dimensional ultrastructure ofCryptococcus neoformans was studied by quick-freezing and deep-etching (QF-DE) method.C. neoformans, strain CDC551, was cultured on agar. The viable yeast cells (107 cells) were inoculated into each mouse from the tail vein. Three weeks after the inoculation, the brains of the mice were perfused with fixatives, quickly frozen, freeze-fractured, deeply etched and rotary shadowed with platinum and carbon. In addition, the viable cells ofC. neoformans on agar were picked up and quickly frozen, and replica membranes were prepared as described above. The ultrastructure ofC. neoformans was three-dimensionally demonstrated by the QF-DE method. The capsule was composed of fine meshworks of microfibrils (10–13 nm in diameter), which were directly attached to the cell walls. The capsule of the in vivo yeasts (yeast cells in the brain lesion) was thicker than that of the in vitro yeasts (yeast cells on agar culture). At the outer part of the cell wall, a particle-accumulating layer was observed. This layer in vivo was thicker than that in vitro. Occasionally, the yeast cells were ingested by phagocytes in the mouse brain. Although the cytoplasm of such yeast cells was destroyed, the capsular meshworks were well preserved. The ultrastructure of the capsule was the same both in cultured and phagocytized yeasts in the cystic lesions of the brains. This lack of morphological changes of the capsular meshworks suggests that they are resistant to the digestion by phagocytes. This stability of capsular structures may provide one of the important pathogenic factors in cystic lesions byC. neoformans. 相似文献
Gas vesicle formation and buoyancy regulation in Pelodictyon phaeoclathratiforme strain BU1 (Green sulfur bacteria) was investigated under various laboratory conditions. Cells formed gas vesicles exclusively at light intensities below 5 mol · m-2 · s-1 in the stationary phase. No effect of incubation temperature or nutrient limitation was observed. Gas space of gas vesicles occupied always less than 1.2% of the total cell volume. A maximum cell turgor pressure of 330 kPa was determined which is comparable to values determined for cyanobacterial species. Since a pressure of at least 485 kPa was required to collapse the weakest gas vesicles in Pelodictyon phaeoclathratiforme, short-term regulation of cell density by the turgor pressure mechanism can be excluded.Instead, regulation of the cell density is accomplished by the cease of gas vacuole production and accumulation of carbohydrate at high light intensity. The carbohydrate content of exponentially growing cells increased with light intensity, reaching a maximum of 35% of dry cell mass above 10 mol · m-2 · s-1. Density of the cells increased concomitantly. At maximum density, protein and carbohydrate together accounted for 62% of the total cell ballast. Cells harvested in the stationary phase had a significantly lower carbohydrate content (8–12% of the dry cell mass) and cell density (1010–1014 kg · m-3 with gas vesicles collapsed) which in this case was independent of light intensity. Due to the presence of gas vesicles in these cultures, the density of cells reached a minimum value of 998.5 kg · m-3 at 0.5 mol · m-2 · s-1.The cell volume during the stationary phase was three times higher than during exponential growth, leading to considerable changes in the buoyancy of Pelodictyon phaeoclathratiforme. Microscopic observations indicate that extracellular slime layers may contribute to these variations of cell volume. 相似文献
Microbial cell surface glycans in the form of glycolipids and glycoproteins frequently play important roles in cell-cell interaction and host immune responses. Given the likely importance of these surface structures in the survival and pathogenesis of Campylobacter jejuni, a concerted effort has been made to characterise these determinants genetically and structurally since the genome was sequenced in 2000. We review the considerable progress made in characterising the Campylobacter glycome including the lipooligosaccharide (LOS), the capsule and O- and N-linked protein glycosylation systems, and speculate on the roles played by glycan surface structures in the life-cycle of C. jejuni. 相似文献
Summary The lenses of albino rats were studied after freeze-drying with scanning electron microscopic technique.The multilamellated lens capsule is covered by adhering fibers in the equatorial region but looks otherwise almost smooth. The cuboidal, polyhedronal cells on the anterior surface of the lens are characterized by their complex interdigitations and their paucity of extracellular space. The cortical polyhedronal, mostly hexagonal lens fibers have several processes interconnecting neighbouring cells. The plasma membrane appears slightly undulated. It is covered by extracellular material. There is a well-delimited extracellular space between the lens fibers in the cortical layer. The results are discussed in relation to previous studies based upon light and electron microscopy.Supported by grants from the Swedish Medical Research Council (B70-12X-2543-02) and Magn. Bergwalls Stiftelse. 相似文献
Abstract Streptococcus suis capsular type 2 has a capsule rich in sialic acid (NANA). Sialic acid, known to be an antiphagocytic factor for many bacterial species, inhibits the activation of the alternative complement pathway. The role of capsular NANA in virulence, resistance to phagocytosis and intracellular survival of S. suis capsular type 2 was evaluated. In general, a low concentration of NANA was observed for all the S. suis strains tested. In addition, no difference could be found in NANA concentrations between strains of different virulence degrees. Sialic acid concentration increased in the virulent strain 89–1591 and the avirulent strain 90–1330 after in vivo growth with an increased capsular material thickness compared to growth in vitro. No significant difference could be found in the phagocytosis rate by porcine blood monocytes of either strain and strain 89–1591 treated with sialidase or the sialic acid-binding lectin from Sambucus nigra (SNA I). Intracellular survival of strain 89–1591 decreased after treatments with sialidase or lectin, becoming comparable to that of strain 90–1330. Finally, no difference could be seen in virulence using a murine model, even if strain 89–1591 was treated with the enzyme or the lectin. Thus, NANA does not seem to be a critical virulence factor for S. suis capsular type 2. 相似文献
The capsule polysaccharide-protein-peptidoglycan complex (insoluble in boiling sodium dodecyl sulfate and hot phenol-water) from cell envelopes of Rhodobacter capsulatus St. Louis was characterized. Hydrofluoric, hydrochloric acid or alkaline hydrolysis solubilized the polysaccharide moiety, whereas the protein-peptidoglycan moiety remained insoluble. On treatment of the protein-peptidoglycan moiety with lysozyme, the protein with peptidoglycan-residues bound was solubilized. It showed a single, broad peptide band (Mr=about 17,000) on sodium dodecyl sulfate polyacrylamide gel-electrophoresis. The same protein was obtained by lysozyme digestion (without preceding hydrofluoric or hydrochloric acid treatment) of the protein-peptidoglycan complex of the phage-resistant mutant Rhodobacter capsulatus St. Louis RC1-, in which the capsule polysaccharide is present in a free form. A protein-peptidoglycan complex was isolated also from the capsulefree Rhodobacter capsulatus 37b4. Covalent binding between the protein and peptidoglycan moieties is likely for all three strains as is the lipoprotein nature of the protein moiety. The polysaccharide moiety of the complete complex from the wild-type Rhodobacter capsulatus St. Louis was at least partly removable from the complex in the presence of high salt concentrations or ethylene diamine tetraacetate. A specific amino acid pattern (with Ser, Gly, Glu, and Ala dominating) remained constantly associated with the capsule polysaccharide moiety independent of the separation procedure.Abbreviations A2pm
diaminopimelic acid
- Cetavlon
cetyltrimethyl-ammonium bromide
- EDTA
ethylene-diaminetetraacetate, disodium salt
- HF
hydrofluoric acid
- HPLC
high-performance liquid chromatography
- PAGL
polyacrylamide gel-electrophoresis
- SDS
sodium dodecyl sulfate
- TCA
trichloroacetic acid 相似文献
This light and electron microscope study reveals considerable uniformity in hornwort stomata morphology and density in contrast to common spatial and developmental abnormalities in tracheophytes and mosses. Stomata arise from a median longitudinal division of sporophyte epidermal cells morphologically indistinguishable from their neighbours apart from the retention of a single chloroplast whilst those in the other epidermal cells fragment. Chloroplast division and side-by-side repositioning of the two daughter chloroplasts determines the division plane in the stomatal mother cell. The nascent guard cells contain giant, starch-filled chloroplasts which subsequently divide and, post aperture opening, regain their spherical shape. Accumulation of wall material over the guard cells and of wax rodlets lining the pores follows opening. While the majority of stomata are bilaterally symmetrical those lining the dehiscence furrows display dextral and sinistral asymmetry due to differential expansion of the adjacent epidermal cells.
The ubiquity of open stomata suggests that these never close with the maturational wall changes rendering movement extremely unlikely. These structural limitations, a liquid-filled stage in the ontogeny of the intercellular spaces, and spores already at the tetrad stage when stomata open, suggest that their primary role is facilitating sporophyte desiccation leading to dehiscence and spore dispersal rather than gaseous exchange. Stomata ontogeny and very low densities, like those in Devonian fossils, suggest either ancient origins at a time when atmospheric carbon dioxide levels were much greater than today or a function other than gaseous exchange regulation. We found no evidence for stomatal homology between hornworts, mosses and tracheophytes. 相似文献
Cryptococcus neoformans (C. neoformans) is an opportunistic fungal pathogen that mainly infects immunocompromised individuals such as AIDS patients. Although cell surface receptors for recognition of C. neoformans have been studies intensively, cytoplasmic recognition of this pathogen remains unclear. As an important detector of pathogen infection, inflammasome can sense and get activated by infection of various pathogens, including pathogenic fungi such as Candida albicans and Aspergillus fumigatus. Our present study showed that acapsular C. neoformans (cap59Δ) activated the NLRP3-, but not AIM2-nor NLRC4- inflammasome. During this process, viability of the fungus was required. Moreover, our in vivo results showed that during the pulmonary infection of cap59Δ, immune cell infiltration into the lung and effective clearance of the fungus were both dependent on the presence of NLRP3 inflammasome. In summary, our data suggest that the capsule of C. neoformans prevents recognition of the fungus by host NLRP3 inflammasome and indicate that manipulation of inflammasome activity maybe a novel approach to control C. neoformans infection. 相似文献