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1.
Abstract Monolayers of epithelial cells obtained by culture of isolated secretory alveoli from the bovine mammary gland were used as target cells in bacterial adhesion assays. The ability of two strains of Streptococcus uberis (EF20 and 0140J) to adhere to these cells was examined using scanning electron microscopy (SEM). The cultured monolayers consisted of two types of epithelial cell one of which possessed many microvilli and another which exhibited only sparse or no microvilli. Strain EF20 adhered more readily and in greater numbers to the cells without microvilli (MV) than to cells possessing microvilli (MV+). Strain 0140J also interacted with a greater proportion of MV cells but adhered to both MV and MV+ cell types in similar numbers.  相似文献   

2.
Adhesion of Salmonella dublin to HEp2 epithelial cells   总被引:1,自引:0,他引:1  
Two strains of Salmonella dublin , grown serially in brain heart infusion broth, were motile and produced mannose sensitive (MS) but not mannose resistant (MR) haemagglutinins; grown on phosphate buffered agar, the strains were poorly motile and phenotypically MSHA- MRHA +. In adhesion tests with HEp2 epithelial cells, broth grown bacteria that were motile and MSHA+ MRHA- adhered better than agar grown bacteria that were poorly motile and MSHA- MRHA+. Thus, in adhesion tests with HEp2 epithelial cells, strains of S. dublin behaved like S. typhimurium strains in that their HEp2 cell adhesiveness was associated with motility and production of MSHA.  相似文献   

3.
Aim:  To investigate the nisin Z innocuity using normal human gingival fibroblast and epithelial cell cultures, and its synergistic effect with these gingival cells against Candida albicans adhesion and transition from blastospore to hyphal form.
Methods and Results:  Cells were cultured to 80% confluence and infected with C. albicans in the absence or presence of various concentrations of nisin Z. Our results indicate that only high concentrations of nisin Z promoted gingival cell detachment and differentiation. Determination of the LD50 showed that the fibroblasts were able to tolerate up to 80  μ g ml−1 for 24 h, dropping thereafter to 62  μ g ml−1 after 72 h of contact, compared to 160  μ g ml−1 after 24 h, and 80  μ g ml−1 after 72 h recorded by the gingival epithelial cells which displayed a greater resistance to nisin Z. The use of nisin Z even at low concentration (25  μ g ml−1) at appropriate concentrations with gingival cells significantly reduced C. albicans adhesion to gingival monolayer cultures and inhibited the yeast's transition.
Conclusion:  These findings show that when used at non-toxic levels for human cells, nisin Z can be effective against C. albicans adhesion and transition and may synergistically interact with gingival cells for an efficient resistance against C. albicans .
Significance and Impact of the Study:  This study suggests the potential usefulness of nisin Z as an antifungal agent, when used in an appropriate range.  相似文献   

4.
The reaggregation kinetics of frog ( Xenopus laevis ) cells prepared using several different dissociation procedures were monitored. Two distinct modes of cell adhesion were revealed, one mediated by Ca2+ dependent cell-cell adhesion system (CDS) and the other mediated by Ca2+ independent one (CIDS).
CDS was detected in frog embryonic (two-cell stage to neurula) cells and in cells of epithelial cell lines (A6 and A8), while CIDS was detected only in A6 cells. Frog CDS was resistant to 0.01% tryptic digestion in the presence of 1 mM Ca2+ and CIDS was resistant to dilute (0.0001%) tryptic digestion. Cells with both mechanisms were prepared by dissociation with 1 mM EDTA and both mechanisms were absent in cells dissociated with 0.01% trypsin and 1 mM EDTA. Frog CDS and CIDS functioned in a temperature dependent and independent manner, respectively. Properties of frog CDS and CIDS revealed in this study were the same as those of mammalian or avian CDS and CIDS, respectively. Frog cells with CDS cross-adhered to mammalian epithelial (F9) cells but not to fibroblastic (V79) cells Ca2+ dependently. Monoclonal antibody ECCD-1 raised against mouse epithelial CDS blocked aggregation of frog A8 cells.
These results suggest the similarity between frog CDS and mammalian epithelial type CDS.  相似文献   

5.
Abstract In contrast to the phorbol ester oxidative response, which only develops during dimethyl-sulphoxide (DMSO)-induced differentiation of the human leukemic myeloblast HL-60 cell-line, the endotoxin response was observed in undifferentiated and differentiated cells. The Ca2+ response to endotoxin, detected in both differentiated and undifferentiated HL-60 cells, consisted of a transient 10–50 nM increase in intracellular Ca2+. A very slow, irreversible increase in intracellular Ca2+ was detected at high 1–100 μg/ml endotoxin concentrations, and this effect, and the inositol phosphate response, correlated with the surfactant activities of various endotoxins and Lipid A. Arachidonic acid and sodium arachidonate 1–50 μM stimulated a large 200–500 nM and transient Ca2+ response in undifferentiated HL-60 cells, which was significantly greater than that elicited by 1–50 μM eicosapentaenoic acid, and was not observed at similar concentrations of arachidonic acid methyl ester or myristic acid. These concentrations (1–50 μM) of arachidonic acid were observed to have surfactant activities on the plasma membrane. At lower arachidonic acid concentrations a marked potentiation of both Ca2+ and oxidative responses to the chemotactic peptide fMet-Leu-Phe was detected. It is possible that the arachidonic acid released during phospholipase A2 activation of neutrophils may be involved in cellular cross-talk and, at higher concentrations, in directly activating Ca2+ and superoxide production. It is also possible that previously reported effects of endotoxin at high concentrations are an vitro artefact of surfactant properties of endotoxin.  相似文献   

6.
Abstract: It is widely accepted, although never demonstrated, that the formation of a disulfide bond in the majority of immunoglobulin (Ig)-like domains stabilizes their final conformation and thus is essential to their functioning as adhesion/recognition molecules. The myelin P0 protein, which has been shown directly to behave as a homophilic adhesion molecule, contains a single Ig-like domain, stabilized by a putative Cys21-Cys98 disulfide bond. To test if this bond is indeed necessary to the adhesive function of P0, the nucleotides in the P0 cDNA coding for Cys21 were altered to code for an alanine. The mutated P0 cDNA was transfected into Chinese hamster ovary cells, expression of the mutated P0 protein was characterized, and the adhesiveness of Cys21-mutated P0-expressing cells and that of cells expressing equivalent surface amounts of the unmutated protein were compared. It was found, as we previously reported, that incubation of a single cell suspension of the unmutated P0-expressing cells resulted in the rapid formation of large aggregates. In contrast, after a similar incubation the cells expressing the Cys21-mutated P0 were still mostly single cells, a result indistinguishable from that observed with the control transfected cells. This suggests that the P0 protein, when mutated at Cys21, does not behave as a homophilic adhesion molecule, which in turn implies that the formation of an Ig domain disulfide bond is essential to the functioning of this molecule.  相似文献   

7.
Mesophyll cells isolated from Phaseolus vulgaris and Lycopersicon esculentum show decreasing photosynthetic rates when suspended in media containing increasing concentrations of osmoticum. The photosynthetic activity was sensitive to small changes in osmotic potential over a range of sorbitol concentrations from 0.44 M (−1.08 MPa) to 0.77 M (−1.88 MPa). Photorespiration assayed by 14CO2 release in CO2-free air and by 14CO2 release from the oxidation of [1–14C] glycolate also decreased as the osmotic potential of the incubation medium was reduced. The CO2 compensation points of the cells increased with increasing concentration of osmoticum from approximately 60 μ I−11 at −1.08 MPa to 130 μl 1−1 for cells stressed at −1.88 MPa. Changes in photosynthetic and photorespiratory activities occurred at moderate osmotic potentials in these cells suggesting that in whole leaves during a reduction in water potential, non- stomatal inhibition of CO2 assimilation and glycolate pathway metabolism occurs simultaneously with stomatal closure.  相似文献   

8.
Epithelial cells are the initial sites of host invasion by group A Streptococcus pyogenes (GAS), and their infection of epithelial cells has been suggested to induce apoptosis. However, the mechanism responsible for bacteria–host interaction and the induction of apoptosis has not been clearly understood. We demonstrate here that human pharyngeal epithelial HEp-2 cells became apoptotic with DNA fragmentation by invasion of GAS strains JRS4 (M6+, F1+) and JRS145 (M6, F1+ mutant of JRS4), whereas apoptotic cellular changes were not observed in SAM1 (M6+, F1 mutant) or SAM2 (M6, F1 mutant) infected HEp-2 cells. Confocal microscopy revealed that Bax translocation to mitochondria and cytochrome c release occurred after 4 h of infection. Western blot analyses showed that the amounts of Bcl-2 and Bcl-xL were decreased in the mitochondria of infected cells. In addition, we demonstrated that the release of nuclear histone from infected cells was prevented by the addition of caspase-9 inhibitor (Ac-LEHD-CHO). We conclude that the internalization of GAS in epithelial cells is necessary and sufficient for the induction of apoptosis, which is initiated by mitochondrial dysfunction, and the mechanism of GAS-induced apoptosis is clearly different from that induced by other intracellular invasive bacteria, e.g. Shigella and Salmonella species.  相似文献   

9.
Primary productivity of algal epiphytes on the surfaces of Phragmites, Potamogeton , and Nuphar was measured seasonally from June 1978, through June 1979, in the littoral zone of Lake Ohrid, using 14C methodology. Surface areas of individual macrophytes were determined throughout the study period through the use of a non-miscible surfactant and a calibration curve of surfactant weight versus known, calculated surface areas.
Mean total surface area available for epiphytic colonization during the study period was 1.032 m2 macrophyte surface per m2 of littoral zone for Phragmites , 0.810 m2 for Potamogeton , and 0.167 m2 for Nuphar . Seasonal rates of mean primary productivity of algal epiphytes on Phragmites from the surface to the light-compensation depth ranged from 84–1406 mg C m−2 littoral zone d−1; ranges for epiphytes on Potamogeton and Nuphar were 77–586 and 69–268 mg C m−2 littoral zone d−1, respectively. Maximum rates were observed typically during June; minimum rates were observed typically during August to December. Mean daily productivity rates over the 12 month period were for epiphytes on Potamogeton 167.0, on Nuphar 100.4 and on Phragmites 671.2 mg C m−2 littoral zone d−1. Calculated annual production for epiphytes on Nuphar was 36.65, on Potamogeton 60.95 and on Phragmites 245.0 g C m−2 littoral zone yr−1. Epiphytic production data were typically considerably higher than production data obtained for littoral and pelagial planktonic algae and compare favorably with published data for epiphytic and periphytic production in Lawrence Lake, Marion Lake, and Borax Lake.  相似文献   

10.
Rapidly dividing photoautotrophic cell suspensions from Chenopodium rubrum L. assimilated about 85 μmol CO2 (mg chlorophyll)−1 h−1. During the late stationary phase of culture growth, CO2 fixation rate was reduced to about 60 μmol CO2 (mg chlorophyll)−1 h−1. Actively dividing cells characteristically incorporated a smaller proportion of 14C into starch than cells from non-dividing stationary phases. In rapidly dividing cells, [14C]-turnover from free sugars, sugar-phosphates, organic and amino acids was substantially higher compared to non-dividing cells from stationary growth phase. Higher proportions of photosynthetically fixed carbon were channelled into proteins, lipids and structural components in actively dividing cells than in non-dividing cells. In the latter. 14C was preferentially channeled into starch, and a striking increase in starch accumulation was observed. The transfer of non-dividing, stationary growth-phase cells into fresh culture medium resulted in an increase in the maximum extractable activities of some enzymes involved in the glycolytic and dark respiratory pathways and in the citric acid cycle. In contrast, the maximum extractable activities of the chloroplastic enzymes, ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.38) and NADP+-glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.13) were highest after the cells had reached the stationary growth phase.  相似文献   

11.
In this study, we characterize the interaction between non-piliated (P) Neisseria gonorrhoeae and human epithelial cells. P mutants lacking the pilus subunit protein PilE attach at low levels to cells. Although the binding may not lead to heavy inflammatory responses, the interaction between P Neisseria and host cells most probably play a role in colonization and asymptomatic carriage of the pathogen. Here we show that the adherence of P N. gonorrhoeae is blocked by GDP-β-S [guanosine 5'- O -(thio)diphosphate], a non-hydrolyzable GTP analogue, and by C3 exotoxin, an inhibitor of the small G-protein Rho. G-protein activators such as cholera toxin, that activates Gs, and fluoroaluminate, a general G-protein activator, induced bacterial adherence. Furthermore, increase of the extracellular free [Ca2+] dramatically enhanced adherence of non-piliated Neisseria . The pharynx and the urogenital tract are natural entry sites of the pathogenic Neisseria species, and at both sites the epithelial cells can be exposed to wide variations in Ca2+ concentration. Taken together, these data show the importance of extracellular Ca2+ in the pathogenic Neisseria -host interaction, and reveal a novel function of cholera toxin, namely induction of bacterial adherence.  相似文献   

12.
Abstract. For Chlorella emersonii , plausible membrane potentials between –80 and –120 mV were calculated from the distribution of the lipophilic cation tetraphenylphosphonium (TPP+) between the cells and the medium. Furthermore, these calculated membrane potentials were influenced in a way expected from the literature, by different metabolic conditions induced by light or dark, anaerobiosis, glucose, and by inhibition or uncoupling of electron transport.
Nevertheless, the experiments presented here indicate that TPP+ is unsuitable as a probe for electrical potentials, at least in Chlorella emersonii. The reasons for this conclusion are as follows:
  • 1. 

    Much of the incorporated TPP+-14C could not be exchanged against unlabelled TPP+.

  • 2. 

    The uptake of TPP+-14C was very slow and exhibited complex rather than simple saturation kinetics.

  • 3. 

    A large adsorption of TPP+-14C took place even after the cells were killed; the adsorption by living cells was only 20–60% higher than with killed cells. Furthermore, the adsorption by killed cells showed kinetics similar to living cells.

  相似文献   

13.
Abstract The reversibility of adhesion of 3 representative strains of oral streptococci from a phosphate-buffered suspension onto 5 different solid substrata was studied.
Streptococcus mitis T9 (surface free energy γb= 39 mJ · m−2). Streptococcus sanguis CH3 (γb= 95 mJ · m−2) and Streptococcus mutans NS (γb= 117 mJ · m−2) were selected on basis of their surface free energy. Solid substrata were employed with a surface free energy γs ranging from 20 mJ · m−2 for polytetrafluorethylene to 109 mJ · m−2 for glass. Bacterial suspensions containing 2.5 × 109 cells per ml were incubated with 2 samples of each substratum. After 1 h the number of adhering bacteria was evaluated on one sample, while the second sample was kept for another hour at a 10-fold lower bacterial concentration. Bacteria with a low surface free energy desorbed only from substrata with a high surface free energy, while bacteria with a high surface free energy desorbed from substrata with a low surface free energy. Thus low energy bacterial strains adhered reversibly to high energy substrata and vice versa. Similar observations were made with polystyrene particles. Calculation of the interfacial free energy of adhesion (Δ F adh) for each bacterial strain as well as for the polystyrene particles showed that a reversible adhesion was associated with a positive Δ F adh, denoting unfavourable adhesion conditions upon a thermodynamic basis.  相似文献   

14.
Abstract We have recently demonstrated that the calmodulin antagonist trifluoperazine has antitubercular activity in vitro against Mycobacterium tuberculosis H37Rv susceptible and resistant to isoniazid. It is shown that trifluoperazine at a concentration of 50 μ g ml−1 when added to the cells along with the labelled precursors inhibited the incorporation of [14C]acetate into lipids (63%) and uptake of [14C]glycine (74%) and [3H]thymidine (52%) bu whole cells of M. tuberculosis H37Rv by 6 h of exposure. After 48 h, the inhibition was 87%, 97% and 74%, respectively. However, when the drug was added to cells taking up and metabolizing the labelled precursors at a later point (3 h for [14C]acetate and [3H]thymidine and 12 h for [14C]glycine) it inhibited completely the uptake of all the precursors, at least up to 24 h. The onset of inhibitory action was very rapid, i.e. 3 h. It is suggested that trifluoperazine has multiple sites of action and acts probably by affecting the synthesis of lipids, proteins and DNA.  相似文献   

15.
The rate of degradation of n -alkanes C12-C18, in petrol (Slovene diesel) in an aqueous system, by free and immobilized Pseudomonas fluorescens in shaking flasks was investigated. Cells were immobilized to a biosupport, Biofix, and a biosorbant, Drizit. Analysis of cellular growth of the free and immobilized bacteria over 8 d of incubation with diesel as the sole carbon source, showed a reduction in the lag phase in the immobilized cultures in comparison to the free system. The free system degraded 52·3% of C12 and 11·6% of C13, but C14-C18 were not degraded. In comparison to the free system and diesel which had not been exposed to experimental conditions (unexposed), the immobilized systems degraded significantly more of C13-C18. Biofix-immobilized cells degraded 14·8% of C12 and an average of 53·5% of C13-C18. Drizit-immobilized cells degraded 24·5% of C12, 52·4% of C13 and an average of 91·2% of C14-C18. This study shows the successful use of immobilized bacteria technology to enhance the degradation of diesel in an aqueous system.  相似文献   

16.
Windowpane flounder, Scophthalmus aquosus Mitchill, were exposed for 60 days to 5 or 10 μg 1−1 mercury and gill samples were examined by scanning electron microscopy. The response of the gill epithelium was different at the two levels of mercury exposure. The number of chloride cell apical pits and gill filaments bearing 'cratered' epithelial cells increased at the 5-μg 1−1 level and decreased at the higher exposure level.
Focal swellings demonstrated a dose-dependent relationship, their numbers being greatest at the higher exposure level. Marked fragmentation of pavement cell microridge patterns and swelling of the respiratory epithelial cells was evident at the 10-μg 1−1 exposure level.  相似文献   

17.
The purinergic receptor P2X7 is involved in cell death, inhibition of intracellular infection and secretion of inflammatory cytokines. The role of the P2X7 receptor in bacterial infection has been primarily established in macrophages. Here we show that primary gingival epithelial cells, an important component of the oral innate immune response, also express functional P2X7 and are sensitive to ATP-induced apoptosis. Porphyromonas gingivalis, an intracellular bacterium and successful colonizer of oral tissues, can inhibit gingival epithelial cell apoptosis induced by ATP ligation of P2X7 receptors. A P. gingivalis homologue of nucleoside diphosphate kinase (NDK), an ATP-consuming enzyme, is secreted extracellularly and is required for maximal suppression of apoptosis. An ndk -deficient mutant was unable to prevent ATP-induced host-cell death nor plasma membrane permeabilization in the epithelial cells. Treatment with purified recombinant NDK inhibited ATP-mediated host-cell plasma membrane permeabilization in a dose-dependent manner. Therefore, NDK promotes survival of host cells by hydrolysing extracellular ATP and preventing apoptosis-mediated through P2X7.  相似文献   

18.
Abstract The prostaglandin response of mouse intestinal epithelial cells after exposure to Escherichia coli heat-stable enterotoxin II was examined. The quantity of prostaglandin E2 produced by the intestinal cells was directly related to the dose of heat-stable enterotoxin II. The change in the amount of prostaglandin E2 over time correlated to that of the volume of fluid released into the intestinal lumen. We then demonstrated that administration of heat-stable enterotoxin II into the intestinal loops of mice induced elevation of arachidonic acid and phosphatidic acid levels in intestinal epithelial cells. These results show that heat-stable enterotoxin II stimulates arachidonic acid metabolism in intestinal epithelial cells and that the synthesized prostaglandin E2 functions as a mediator of fluid secretion induced by this enterotoxin.  相似文献   

19.
Neisseria gonorrhoeae has a repertoire of up to 11 opacity-associated (Opa) proteins that are adhesins. Most Opa proteins adhere to CEACAM antigens and when CEACAM molecules are present on the surface of transfected epithelial cells their binding by Opa is thought to induce invasion of these cells by gonococci. In this study, we investigated whether several malignant epithelial cell lines, normal cervical and fallopian tube epithelial cell cultures, as well as normal fallopian tube tissue express several of the CEACAM molecules, and whether gonococci use these molecules for adherence and invasion of these female genital epithelial cells. A primary cervical cell culture and metastatic cervical cell line ME180 both expressed CEACAM as shown by whole cell ELISA and flow cytometry, and increased the surface expression of total CEACAM during incubation with Opa+ gonococci. Opa+ gonococci both adhered to and invaded these cells; CEACAM-specific monoclonal antibody (MAb) partially abolished this interaction. Two primary fallopian epithelial tube cell cultures, a primary cervical cell culture and two malignant cell lines, HEC-1-B and HeLa, did not express CEACAM nor was CEACAM mRNA present. No evidence of either intracellular or secreted extracellular CEACAM was found with HEC-1-B and HeLa cells. Opa+ gonococci both adhered to and invaded CEACAM non-expressing cells; however, Opa+ gonococcal association with these non-expressing cell lines could not be inhibited with CEACAM-specific MAb. These data show that CEACAM is not always expressed on female genital epithelial cells and is not essential for gonococcal adherence and invasion. However, when CEACAM is expressed, Opa+ gonococci exploit it for the adherence to and invasion of these cells.  相似文献   

20.
Abstract: Three isoforms of catalytic α subunits and two isoforms of β subunits of Na+,K+-ATPase were detected in rat sciatic nerves by western blotting. Unlike the enzyme in brain, sciatic nerve Na+,K+-ATPase was highly resistant to ouabain. The ouabain-resistant α1 isoform was demonstrated to be the predominant form in rat intact sciatic nerve by quantitative densitometric analysis and is mainly responsible for sciatic nerve Na+,K+-ATPase activity. After sciatic nerve injury, the α3 and β1 isoforms completely disappeared from the distal segment owing to Wallerian degeneration. In contrast, α2 and β2 isoform expression and Na+,K+-ATPase activity sensitive to pyrithiamine (a specific inhibitor of the α2 isoform) were markedly increased in Schwann cells in the distal segment of the injured sciatic nerve. These latter levels returned to baseline with nerve regeneration. Our results suggest that α3 and β1 isoforms are exclusive for the axon and α2 and β2 isoforms are exclusive for the Schwann cell, although axonal contact regulates α2 and β2 isoform expressions. Because the β2 isoform of Na+,K+-ATPase is known as an adhesion molecule on glia (AMOG), increased expression of AMOG/β2 on Schwann cells in the segment distal to sciatic nerve injury suggests that AMOG/β2 may act as an adhesion molecule in peripheral nerve regeneration.  相似文献   

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