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1.
Background: Human β-defensin 2 (hBD2) gene expression is dependent on nuclear factor kappa B (NF-κB) activity. We have previously demonstrated that electrolytically generated acid functional water (FW) induces the expression of hBD2 in the human oral squamous cell carcinoma (OSCC) cell line Ca9-22. However, the induction was not dependent on NF-κB activity; in fact, FW inhibited NF-κB activity. Therefore, we hypothesized that FW might reduce spontaneous interleukin 8 (IL-8) secretion by Ca9-22 cells, which is heavily dependent on NF-κB activity. This study aimed at demonstrating the inhibitory effect of FW on NF-κB activity. Methods: Ca9-22 cells were incubated with FW, and spontaneous IL-8 secretion was observed by enzyme-linked immunosorbent assay. Luciferase assay was performed using the 5′-untranslated region of the IL-8 gene. The steps of NF-κB activation blocked by FW were evaluated by localization of the NF-κB subunits p65 and p50 by immunofluorescence staining. Western blotting was further performed to confirm the changes in NF-κB subunit localization. Results: The Ca9-22 cells spontaneously secreted IL-8, which was rapidly and drastically inhibited by FW treatment. The luciferase assay demonstrated the inhibitory action of FW, which was diminished by deletion of the NF-κB binding site from this construct. FW treatment altered the distribution of both the p65 and p50 subunits. P65, which was localized in the nucleus during the resting state, moved to the cytoplasm after FW treatment, whereas, p50, localized in the cytoplasm during the resting state, moved to the nucleus subsequent to FW treatment. Conclusions: The results from this study indicate that FW might inhibit spontaneous IL-8 secretion by redistribution of the NF-κB subunits within the cells.  相似文献   

2.
Systematic isolation of the cell constituents of Proteus mirabilis RMS–203 was performed to find out localization of antitumor principle only in the lipopolysaccharide (LPS) layer of the cell wall fraction.

LPS with strong antitumor activity was extracted from P. mirabilis RMS–203 by phenol-water method followed by purification on DEAE-Sephadex A–50 column chromatography.

The main components of purified LPS were galactose, hexosamine, 2-keto-deoxy-octonic acid (KDO), myristic acid, β-hydroxymyristic acid and α,ε-diaminopimelic acid.

The minimal effective dose of LPS against Ehrlich solid carcinoma in mice was 0.1~1.0 μg/mouse. LD50 in mice and pyrogenicity in rabbits were 28 mg/kg and 10?3–10?5 μg/rabbit, respectively.  相似文献   

3.
In order to elucidate the interaction of lactic acid bacteria with the immune system, immune responses to the lactic acid bacteria, Bifidobacterium longum and Lactobacillus acidophilus, were examined in mice fed with each organism. In mice fed with B. longum for more than 8 weeks, an antibody response was detected to the cytoplasm of B. longum, but not to the cell wall. On the other hand, in mice fed with L. acidophilus for more than 6 weeks, an antibody response was detected to both the cytoplasm and cell wall of L. acidophilus. Moreover, feeding each organism for 2 weeks enhanced the proliferative response of Peyer’s patch (PP) cells to the cell fraction against which the serum antibody was detected. However, this was not found with spleen cells. These results suggest that mucosal stimulation by lactic acid bacteria may induce a systemic immune response to them.  相似文献   

4.
The production of microbial cell substances from hydrocarbons has been attracting attention of people for many years. Production of bacterial cell from hydrocarbons is disadvantageous because of the difficulty in separating cell from the broth.

We have tested hydrocarbon-utilizing yeasts isolated from garden soil for cell production. The effect of medium composition on yeast growth and the utilization of individual hydrocarbon by yeast, strain Y-3, were investigated.

As a nitrogen source, urea was more effective than ammonium nitrate. When a very smal! amount of corn steep liquor was added, yeast growth was very improved. Aliphatic series of hydrocarbon lower than C9 were not or very slightly assimilated by this yeast.

Generally speaking, series of even-number hydrocarbons were more effective than those of odd-number hydrocarbons.

We found that the yeast Y-3 strain reported in the previous paper1) has a diterminal oxidation system of hydrocarbon.

This yeast capable of growing in mineral-salts solution with hydrocarbons as sole source of carbon produced a series of dioic acid from n-undecane. These acids are 1,11-undecane dioic acid, 1,9-nonane dioic acid (azelaic acid), 1,7-heptane dioic acid (pimelic acid) and 1,5-pentane dioic acid (glutaric acid). 1,10-Decane dioic acid (sebacic acid) was also isolated from n-decane cultures.

Azelaic acid was partially transformed into pimelic acid and glutaric acid by treating it with resting cells of this yeast.

1,11-Undecane dioic was also transformed into azelaic acid pimelic acid, and glutaric acid by the same treatment as described above.  相似文献   

5.
Abstract

Vitamin B12 and propionic acid that were simultaneous produced by Propionibacterium freudenreichii are both favorable chemicals widely used in food preservatives, medicine, and nutrition. While the carbon source and propionic acid accumulation reflected fermentation efficiency. In this study, using corn stalk as a carbon source and fed-batch fermentation process in an expanded bed adsorption bioreactor was studied for efficient and economic biosynthesis of acid vitamin B12 and propionic. With liquid hot water pretreated corn stalk hydrolysates as carbon source, 28.65?mg L?1 of vitamin B12 and 17.05?g L?1 of propionic acid were attained at 168?h in batch fermentation. In order to optimize the fermentation outcomes, fed-batch fermentation was performed with hydrolyzed corn stalk in expanded bed adsorption bioreactor (EBAB), giving 47.6?mg L?1 vitamin B12 and 91.4?g L?1 of propionic acid at 258?h, which correspond to product yields of 0.37?mg g?1 and 0.75?g g?1, respectively. The present study provided a promising strategy for economically sustainable production of vitamin B12 and propionic acid by P. freudenreichii fermentation using biomass cornstalk as carbon source and expanded bed adsorption bioreactor.  相似文献   

6.

In the present work, a positive effect was obtained by using low molecular weight chitosan compounds in combination with synthetic fungicides. Antifungal activity against Botrytis cinerea, determined by the radial growth method, was more than 75%, with a 25?×?10??10 g/L concentration of fludioxonil or difenoconazole in compounds. Metabolic activity of B. cinerea fungus was about 15% when using a chitosan compound containing fludioxonil at a concentration of 25?×?10??7 g/L. The combined action of chitosan with difenoconazole at a fungicide concentration of 25?×?10??4 g/L is 2–3 times more effective than the action of each component separately. Results of studies for artificially inoculated B. cinerea tomato fruit when treated with low molecular chitosan and chitosan conjugate with gallic acid reduced the frequency of rotting fruit by 50 and 83%, respectively. Chitosan-gallic acid conjugate were obtained from chitosans with Mw of 28 kDa (Ch28GA) was proved to be effective as a preventive treatment for 3 days and can potentially be used as a biofungicide against B. cinerea on tomatoes in the post-harvest period.

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7.
A column of insoluble concanavalin A was prepared by coupling the protein to cyanogen bromide-activated Sepharose. When autolysates of Bacillus subtilis 168 cell walls were passed over the column, the alpha glucosylated teichoic acid component of the cell wall was retained. The teichoic acid could be eluted with dilute alpha-methylglucopyranose. The teichoic acid prepared by affinity chromatography from cell wall autolysates had a higher sedimentation rate than teichoic acids obtained by conventional methods.

Several authors have shown that concanavalin A (con A) forms complexes with alpha-glucosylated teichoic acids1–3. Doyle and Birdsell1 found that the teichoic acid of Bacillus subtilis 168 (trp C2) would precipitate with con A at neutral pH in dilute buffer. The formation of a precipitate was inhibited by sugars which bind to the active site of con A. This observation suggested that it should be possible to purify the teichoic acid by affinity chromatography using insoluble con A as the affinity probe. Lloyd4 and Donnelly and Goldstein5 have successfully employed insoluble con A to purify polysaccharides and glycoproteins. In this communication, we describe conditions for the rapid purification of the alpha-glucosylated teichoic acid of B. subtilis 168. The teichoic acid prepared by this procedure appears to be less degraded than teichoic acids obtained by conventional methods.  相似文献   

8.
Abstract

The effect of sugars on growth of isolated cotyledons from germinating castor bean seeds. — The presence of sugars in the medium induces a number of biochemical and physiological changes in the cotyledons isolated from germinating castor bean seeds. In the present research, the extent and the mechanism of the effect of sugars on growth of the isolated cotyledons was investigated.

The results achieved may be summarized as follows:

  • Sucrose markedly stimulates the growth (as increase of fresh weight) of the isolated cotyledons; its action is already apparent when using a 10-2M concentration and reaches a maximum (stimulation by about 400%) for the concentration between 5.10-2M and 1,5.10-3M. The increase of sucrose induced growth is due in almost equal proportions to water uptake and to increase of dry weight.

  • The difference (Δ P.O.) between the osmotic pressure (P.O.) of the cell sap and that of the external medium is markedly higher for the cotyledons treated with sucrose (7–8 atm.) than the Δ P.O. of cotyledons incubated in water.

  • Analyses of cell contents in sugars show that sucrose is taken up by the cotyledon cells against a concentration gradient. The increase of the difference of osmotic pressure between cotyledon and the external medium is satisfactorily accounted for by this active accumulation of sugars.

  • When mannitol is added to the incubating medium in addition to sucrose, the active uptake of sugar is not disturbed; but the effect of sucrose on growth decreases, and it completly disappears when mannitol concentration in the medium is such, to make the value of Δ P.O. for the cotyledons in sucrose plus mannitol equal to the Δ P.O. for the cotyledons incubated in water.

  • Besides increasing the P.O. in the cotyledons, sucrose markedly accelerates the decrease of free aminoacid in the isolated cotyledons.

  • Auxin (β—indolacetic acid) does not stimulate, or stimulates weakly, growth of the cotyledon incubated in water; some stimulation can be observed only when sucrose is present. Gibberellic acid stimulates growth (though to a much lower extent than sucrose) in the cotyledons in water, while its action does no longer appear when sugar is present.

The decrease of free aminoacid together with other observations, indicates an effect of sucrose on protein metabolism; however, growth stimulation by sucrose seems to depend essentially on its active accumulation - against a concentration gradient - and thuson the increase of osmotic pressure. It is thus suggested that in these tissues osmotic pressure is the most important factor in limiting growth by cell extension.  相似文献   

9.
Morphological, histochemical and cytochemical changes were examined in honeybee larvae after infection with the bacterium Bacillus larvae. The results indicate cell necrosis in the midgut epithelium accompanied by increasing cell vacuolization and nuclear pyknosis following per os inoculation with B. larvae. Many autolysosomes were positive for acid phosphatase. Non-vacuolar acid phosphatase activity was also found in lysed cell compartments. No such activity was found in regenerative epithelial cells. Degradation of haemocytes, salivary glands and other tissues was also observed. Histochemical analyses after per cutaneous inoculation with B. larvae of three- and five-day-old honeybee larvae show intense non-vacuolar acid phosphatase activity followed by disintegration of infected salivary glands, epithelial cell cytoplasm and haemocytes.  相似文献   

10.
In P. tetraurelia each cell is determined to express only one of the two complementary mating types, O and E. This determination is under cytoplasmic control and seems to be achieved only by the commitment or noncommitment to the expression of mating type E. All the previously known mutations affecting the differentiation of mating type prevent the expression of the E mating type (O-restricted mutations) without affecting the determination process. An E-restricted mutation was obtained: mtFE. Its phenotypic properties indicate that the mutation affects the determination process itself. When an O cell becomes mtFE/mtFE it acquires the E mating type and an E-determining cytoplasm. We propose that this constitutive determination for the E mating type is due to the inefficiency of a factor which is normally active in an O cell. This factor would act like a repressor and stabilize the E functions under an inactive state.  相似文献   

11.
Abstract

Betulin (B) and betulinic acid (BA) are two triterpenes with diverse pharmacological and physiological actions. Elicitation of Betula pendula Roth cell cultures by elicitors is an excellent strategy to increase B and BA levels. Six abiotic and biotic elicitors were studied to improve accumulation of B and BA in the cell culture of B. pendula. The B and BA production in treated cells was verified by HPLC. The results showed the maximum growth index (7) on day 3 in cells treated with 0.5?mg L?1 chlorocholine chloride (CCC). The increased accumulation of BA in the cells treated with 200?mg L?1 of chitosan was found to be 5.9?×?(6.5?mg g?1 DW) higher over control cells. Treating the cells with 2?mg L?1 of CCC, after 7?days, led to 149.3× enhancement of B content (19.4?mg g?1 DW) over the controls. Production of this triterpenoid at a much shorter time with a much higher growth rate can be economic and lead to producing large amounts of B and BA for anti-cancer and HIV drugs preparation.  相似文献   

12.
A cell‐wall deficient strain of Chlamydomonas reinhardtii P. A Dang. CC‐849 was cotransformed with two expression vectors, p105B124 and pH105C124, containing phbB and phbC genes, respectively, from Ralstonia eutropha. The transformants were selected on Tris‐acetate‐phosphate media containing 10 μg · mL?1 Zeomycin. Upon further screening, the transgenic algae were subcloned and maintained in culture. PCR analysis demonstrated that both phbB and phbC genes were successfully integrated into the algal nuclear genome. Poly‐3‐hydroxybutyrate (PHB) synthase activity in these transgenic algae ranged from 5.4 nmol · min?1 · mg protein?1 to 126 nmol · min?1 · mg protein?1. The amount of PHB in double transgenic algae was determined by gas chromatography–mass spectrometry (GC–MS) when comparing with PHB standard. In addition, PHB granules were observed in the cytoplasm of transgenic algal cells using TEM, which indicated that PHB was synthesized in transgenic C. reinhardtii. Hence, results clearly showed that producing PHB in C. reinhardtii was feasible. Further studies would focus on enhancing PHB production in the transgenic algae and targeting the chloroplast for PHB accumulation.  相似文献   

13.
《Autophagy》2013,9(5):515-517
Larval salivary glands of bees provide a good model for the study of hormone-induced programmed cell death in Hymenoptera because they have a well-defined secretory cycle with a peak of secretory activity phase, prior to cocoon spinning, and a degenerative phase, after the cocoon spinning. Our findings demonstrate that there is a relationship between apoptosis and autophagy during physiological cell death in these larval salivary glands, that adds evidence to the hypothesis of overlap in the regulation pathways of both types of programmed cell death. Features of authophagy include cytoplasm vacuolation, acid phosphatase activity, presence of autophagic vacuoles and multi-lamellar structures, as well as a delay in the collapse of many nuclei. Features of apoptosis include bleb formation in the cytoplasm and nuclei, with release of parts of the cytoplasm into the lumen, chromatin compaction, and DNA and nucleolar fragmentation. We propose a model for programmed cell death in larval salivary glands of Apis mellifera where autophagy and apoptosis function cooperatively for a more efficient degeneration of the gland secretory cells.

Addendum to:

Programmed Cell Death in the Larval Salivary Glands of Apis mellifera (Hymenoptera, Apidae)

E.C.M. Silva-Zacarin, G.A. Tomaino, M.R. Brochetto-Braga, S.R. Taboga, R.L.M. Silva de Moraes

J Biosci 2007; 32:309-28  相似文献   

14.
Using the monoclonal antibody (MoAb) Xa5B6 as probe, the authors examined the mechanisms of cytoplasmic rearrangement occurring during maturation of theXenopus oocyte. The antigen molecules recognized by the MoAb are arranged in radial striations of the oocyte cytoplasm. The radial striations were disorganized in vitro by progesterone treatment, and the antigen molecules were uniformly distributed, predominantly in the animal hemisphere. Even when the germinal vesicle was mechanically removed or when germinal vesicle breakdown was suppressed in a K+-free medium, progesterone induced a disorganization of the radial striations. This progesterone-induced disorganization was inhibited by the protein synthesis inhibitor cycloheximide. When full-sized oocytes were treated with cytochalasin B, the radial striations were also disorganized, but the antigen molecules did not disperse into the large mass. Colchicine treatment had little effect. Antigen molecules were no longer arranged in radial striations and were completely dispersed when the oocyte was simultaneously treated with both drugs. These results indicate that the two compartments in the oocyte cytoplasm, the yolk-free cytoplasm and yolk column, are organized by different types of cytoskeletal system. It is also suggested that the maturation-promoting factor (MPF) activated during progesterone-induced maturation disrupts these cytoskeletal systems and disorganizes the radial striations. Correspondence to: A.S. Suzuki  相似文献   

15.
Dihydrodipicolinate (DDP)* synthetase and DDP-reductase were partially purified about 30 and 15 folds, respectively, from sonic extracts of Brevibacterium flavum.

In contrast with DDP-synthetase from Escherichia coli, the B. flavum enzyme was only slightly inhibited by α, ε-diaminopimelate, a precursor of lysine, but not by lysine itself. Single or simultaneous addition of any other amino acid(s) of aspartate family did not affect the activity significantly. Optimum pH for DDP-synthetase was 8.4 with Tris-HCl buffer. Kms for aspartic-β-semialdehyde and pyruvate at pH 7.5 were 2×10?4m and 1×10?4m, respectively. The formation of DDP-synthetase was not significantly repressed by lysine.

DDP-reductase of B. flavum required NADH or NADPH as the cofactor. This enzyme was not inhibited by single or simultaneous addition of aspartate family amino acid(s).

From the above results, the regulation mechanism of lysine biosynthesis in B. flavum was discussed.  相似文献   

16.
The effect of abscisic acid on growth, ultrastructure and nucleic acid biosynthesis was studied in tissue culture of spinach (Spinacia oleracea L.). Low concentration (0.01 mg l?1) of abscisic acid increased fresh and dry weight of calluses, whereas 1.0 mg l?1 was inhibitory. The stimulating effect was observed only in the presence of a relatively high concentration of kinetin (1 mg l?1). The inhibitory effect was partly overcome by the same kinetin concentration. The low concentration of abscisic acid probably accelerated the induction of callus growth after subculture and stimulated cell division in the exponential phase of growth. Electron microscopy showed the presence of numerous polysomes and rough endoplasmic reticulum in callus cells grown at the stimulating abscisic acid concentration. Control cells and cells at the inhibitory concentration had slightly hyaline cytoplasm and were more vacuolated. Incubation of callus tissue with 32P in the presence of stimulating concentration of abscisic acid showed a significant increase in the rate of biosynthesis of all nucleic acid classes after 8 h, whereas inhibitory concentration produced a decrease in 32P incorporation. However, when the tissue was grown in the presence of abscisic acid for 20 days, both concentrations decreased the rate of nucleic acid biosynthesis, as compared to the controls.  相似文献   

17.

Content of p‐coumaric (PCA) and ferulic (FA) acid was determined by the HPLC method in fourteen forbs with a potential utilization as forages (range of nutrient content per kg DM: 100 to 244gCP, 339 to 528 g NDF and 180–369 g ADF. PCA and FA were determined after methanol extraction in four fractions: free phenolic acids extracted into ether, ester‐bound phenolic acids after alkaline hydrolysis, glycoside‐bound phenolic acids after acid hydrolysis, and cell wall‐bound phenolic acids after alkaline hydrolysis of the solid residue after the extraction with methanol.

Cell wall‐bound phenols were quantitatively the most important fraction (50% of total PCA and 47% of total FA, respectively). The differences among plant species in total PCA plus FA content were significant (F‐value 775, P < 0.01). The range of total phenol content was 31.3 to 416.3 mg/100g DM, the overall mean was 84 mg/100g DM.

Content of phenolic acids was correlated neither with ADF, NDF or ADL content (R 2 = 1–3%, P > 0.05) nor with CP degradability (R 2 = 3% and R 2 = 1% for PCA and FA, respectively, P > 0.05).

95.4% and 30.9% of total PCA, and 98.3% and 72.5% of total FA disappeared in the rumen from the sample of Glechoma hederacea (species with the highest phenol content) and from the sample of Galium aparine (species with low phenol content), respectively, within the four hour incubation interval.

It is presumed that in comparison with grasses, PCA and FA concentration in tested forbs represents a much lower risk in potential ruminant nutrition.  相似文献   

18.
19.
Unfertilized eggs of the newt, Cynops pyrrhogaster, are arrested at the second meiotic metaphase, with activity of the M‐phase promoting factor (MPF) maintained at a high level. After fertilization, the eggs resume the cell cycle, and emit the second polar body. When the change in [Ca2+]i in the fertilized eggs was monitored by aequorin, an early increase in [Ca2+]i was observed 5–10 min after insemination and continued for about 30 sec. A late increase in [Ca2+]i then occurred 10–15 min after fertilization and continued for 30–40 min. The injection of 1,2‐Bis (2 aminophenoxy) ethane‐N,N,N′,N′,‐tetraacetic acid (BAPTA) into unfertilized eggs inhibited reinitiation of the cell cycle after fertilization. Western blot analysis with antibodies against cyclin B1 or Mos indicated that both cyclin B1 and Mos were present in unfertilized eggs, but both disappeared within 30 min after fertilization. Treatment with Ca2+‐ionophore decreased both cyclin B1 and Mos. Chymotryptic activity in Cynops egg extracts was not significantly increased after fertilization or activation by treatment with the Ca2+‐ionophore. No change in [Ca2+]i was observed following treatment with cycloheximide, but the amount of both cyclin B1 and Mos rapidly decreased. These results indicate that resumption of meiosis in Cynops eggs is induced by an increase in [Ca2+]i at fertilization, which causes degradation of both cyclin B1 and Mos by inhibition of de novo synthesis of those proteins. Mol. Reprod. Dev. 53:341–349, 1999. © 1999 Wiley‐Liss, Inc.  相似文献   

20.
Microbial transformations of selenite by methane-oxidizing bacteria   总被引:1,自引:0,他引:1  

Methane-oxidizing bacteria are well known for their role in the global methane cycle and their potential for microbial transformation of wide range of hydrocarbon and chlorinated hydrocarbon pollution. Recently, it has also emerged that methane-oxidizing bacteria interact with inorganic pollutants in the environment. Here, we report what we believe to be the first study of the interaction of pure strains of methane-oxidizing bacteria with selenite. Results indicate that the commonly used laboratory model strains of methane-oxidizing bacteria, Methylococcus capsulatus (Bath) and Methylosinus trichosporium OB3b, are both able to reduce the toxic selenite (SeO3 2?) but not selenate (SeO4 2?) to red spherical nanoparticulate elemental selenium (Se0), which was characterized via energy-dispersive X-ray spectroscopy (EDXS), X-ray absorption near-edge structure (XANES) and extended X-ray absorption fine structure (EXAFS). The cultures also produced volatile selenium-containing species, which suggests that both strains may have an additional activity that can transform either Se0 or selenite into volatile methylated forms of selenium. Transmission electron microscopy (TEM) measurements and experiments with the cell fractions cytoplasm, cell wall and cell membrane show that the nanoparticles are formed mainly on the cell wall. Collectively, these results are promising for the use of methane-oxidizing bacteria for bioremediation or suggest possible uses in the production of selenium nanoparticles for biotechnology.

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