• Restorative Dentistry and Endodontics
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• v.20 no.1
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• pp.71-95
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• 1995

Bacteria have been regarded as a one of major etiologic factors in root canal infections. In endodontic treatment the effective removal of pathogenic microorganisms in the root canal is the key to successful outcome. Bacterial cell wall components may play an important role in the development of pulpal and periapical disease. The purpose of this study was to evaluate the effect of sonic extracts of Streptococcus spp. on cultured L929 cells and to characterize cell wall protein profiles of Streptococcus spp. Streptococcus spp. were isolated from infected root canals and identified with Vitek Systems(Biomeriux, USA). Five streptococci, namely S. sanguis, S. mitis, S uberis, S. mutans (ATCC 10449) and S. faecalis (ATCC 19433) weere enriched in brain heart infusion broth. Cell pellets were sonicated and cell wall extracts were dialyzed and membrane filtered. Prepared cell wall proteins were applied to cultured L929 cell. The cell reaction were evaluated by monitoring DNA synthesis, cell numbers and the change of cell morphology. The total cell wall protein profiles of microorganisms were characterized by sodium dodecyl sulfate polyacrylamide-gel eledruphoresis(SDS-PAGE). DNA synthesis of L929 cells were reduced by the increasing concentration of sonic extracts. DNA synthesis was significantly suppressed in more than $50{\mu}g$/ml of sonic extract conentration in five streptococci. S. nutans (ATCC 10449) showed stronger suppression on DNA synthesis than remaining four streptococci, which had the similar effect on DNA synthesis. Analysis of DNA synthesis measured by [$^3H$]-thymidine uptake was more sensitvie method than cell counting. Sonic extracts affected the microscopic findings of L929 cells. The protein profiles indicated that all five strains shared two major proteins with molecular masses of 70.8 and 57.5 kD respectively. S. uberis and S. mutans shared common minor proteins of which molecular weights were 147.9 and 112.2 kD respectively. However some minor proteins were unique for S. mitis, S. uberis and S. faecalis.

• Proceedings of the Korean Society of Crop Science Conference
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• 2007.04a
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• pp.65-73
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• 2007

The objective of this study is to understand how regulatory mechanisms respond to sugar status for more efficient carbon utilization and source-sink regulation in plants. So, we need to identify and characterize many components of sugar-response pathways for a better understanding of sugar responses. For this end, genes responding change of sugar status were screened using Arabidpsis cDNA arrays, and confirmed thirty-six genes to be regulated by sucrose supply in detached leaves by RNA blot analysis. Eleven of them encoding proteins for amino acid metabolism and carbohydrate metabolism were repressed by sugars. The remaining genes induced by sugar supply were for protein synthesis including ribosomal proteins and elongation factors. Among them, I focused on three hydrolase genes encoding putative $\beta$-galactosidase, $\beta$-xylosidase, and $\beta$-glucosidase that were transcriptionally induced in sugar starvation. Homology search indicated that these enzymes were involved in hydrolysis of cell wall polysaccharides. In addition to my results, recent transcriptome analysis suggested multiple genes for cell wall degradation were induced by sugar starvation. Thus, I hypothesized that enzyme for cell wall degradation were synthesized and secreted to hydrolyze cell wall polysaccharides producing carbon source under sugar-starved conditions. In fact, the enzymatic activities of these three enzymes increased in culture medium of Arabidopsis suspension cells under sugar starvation. The $\beta$-galactosidase encoded by At5g56870 was identified as a secretory protein in culture medium of suspension cells by mass spectrometry analysis. This protein was specifically detected under sugar-starved condition with a specific antibody. Induction of these genes was repressed in suspension cells grown with galactose, xylose and glucose as well as with sucrose. In planta, expression of the genes and protein accumulation were detected when photosynthesis was inhibited. Glycosyl hydrolase activity against galactan also increased during sugar starvation. Further, contents of cell wall polysaccharides especially pectin and hemicellulose were markedly decreased associating with sugar starvation in detached leaves. The amount of monosaccharide in pectin and hemicellulose in detached leaves decreased in response to sugar starvation. These results supported my idea that cell wall has one of function to supply carbon source in addition to determination of cell shape and physical support of plant bodies.

• Journal of Microbiology and Biotechnology
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• v.16 no.2
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• pp.247-255
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• 2006

Yeast cell wall matrix particles are composed entirely of mannoprotein and ${\beta}-glucan$. The mannoproteins of yeast cell wall can systemically enhance the immune system. We previously purified and analyzed alkali-soluble ${\beta}-glucans$ [${\beta}$-(1,3)- and ${\beta}$-(1,6)-glucans] [10]. In the present study, a wild-type strain was first mutagenized with ultraviolet light, and the cell wall mutants were then selected by treatment with 1.0 mg/ml laminarinase (endo-${\beta}$-(1,3)-D-glucanase). Mannoproteins of Saccharomyces cerevisiae were released by laminarinase, purified by concanavalin-A affinity and ion-exchange chromatography. The results indicated that the mutants yielded 3-fold more mannoprotein than the wild-type. The mannoprotein mass of mutant K48L3 was 2.25 mg/100 mg of yeast cell dry mass. Carbohydrate analysis revealed that they contained mannose, glucose, and N-acetylglucosamine. Saccharomyces cerevisiae cell wall components, mannoproteins, are known to interact with macrophages through receptors, thereby inducing release of tumor necrosis factor alpha ($TNF-{\alpha}$) and nitric oxide. Mannoprotein tractions in the present study had a higher macrophage activity of secretion of $TNF-{\alpha}$ and nitric oxide and direct phagocytosis than positive control ($1{\mu}g$ of lipopolysaccharide). In particular, F1 and F3 fractions in mannoproteins of K48L3 enhanced and upregulated the activity of nitric oxide secretion and macrophage phagocytosis by approximately two- and four-fold, respectively.

• Journal of Microbiology and Biotechnology
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• v.32 no.1
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• pp.37-45
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• 2022

The fungal cell wall and membrane are the principal targets of antifungals. Herein, we report that myricetin exerts antifungal activity against Candida albicans by damaging the cell wall integrity and notably enhancing the membrane permeability. In the presence of sorbitol, an osmotic protectant, the minimum inhibitory concentration (MIC) of myricetin against C. albicans increased from 20 to 40 and 80 ㎍/ml in 24 and 72 h, respectively, demonstrating that myricetin disturbs the cell wall integrity of C. albicans. Fluorescence microscopic images showed the presence of propidium iodide-stained C. albicans cells, indicating the myricetin-induced initial damage of the cell membrane. The effects of myricetin on the membrane permeability of C. albicans cells were assessed using crystal violet-uptake and intracellular material-leakage assays. The percentage uptakes of crystal violet for myricetin-treated C. albicans cells at 1×, 2×, and 4× the MIC of myricetin were 36.5, 60.6, and 79.4%, respectively, while those for DMSO-treated C. albicans cells were 28.2, 28.9, and 29.7%, respectively. Additionally, myricetin-treated C. albicans cells showed notable DNA and protein leakage, compared with the DMSO-treated controls. Furthermore, treatment of C. albicans cells with 1× the MIC of myricetin showed a 17.2 and 28.0% reduction in the binding of the lipophilic probes diphenylhexatriene and Nile red, respectively, indicating that myricetin alters the lipid components or order in the C. albicans cell membrane, leading to increased membrane permeability. Therefore, these data will provide insights into the pharmacological worth of myricetin as a prospective antifungal for treating C. albicans infections.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.7
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• pp.935-940
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• 2001

To determine the effect of different growth stages and cultivars on the chemical composition of sorghum plant and its morphological fractions, samples of whole plant, leaf and stem of J.S-263, J.S-88 and Hegari cultivars, harvested at various growth stages were drawn for analysis. All the samples were analysed for their dry matter contents and various cell wall components such as NDF, ADF. hemicellulose, cellulose, lignin, cutin and silica. Significant increase in DM contents of whole sorghum plant, leaf and stem was observed with advancing stage of growth. The highest DM content was recorded in leaf fraction of the plant. All the cell wall constituents increased significantly in whole sorghum plant, leaf and stem as the plant matured. The maximum NDF, ADF, cellulose and lignin contents were observed in stem fraction, followed by whole plant. However, the hemicellulose, cutin and silica contents were higher in leaf fraction of the plant. The cultivars were found to have some effect on the chemical composition of whole plant, leaf and stem fractions. The results indicated that plant maturity had a much greater effect on the chemical composition of sorghum plant, whereas it was little affected by cultivars.

• Asian-Australasian Journal of Animal Sciences
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• v.12 no.3
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• pp.366-370
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• 1999

Samples of whole plant, leaf and stem of Akbar, Neelum, UM-81 and lZ-31 cultivars of maize fodder harvested up to 14 weeks at different growth stages were drawn and analysed for dry matter contents and various cell wall constituents such as NDF, ADF, hemicellulose, cellulose, lignin, cutin and silica. The dry matter contents of whole maize plant, leaf and stem increased significantly (p<0.01) with advancing plant age. Maximum dry matter was found in the leaf fraction of the plant. The cell wall components continued to increase significantly (p<0.001) in whole maize plant and its morphological fractions as the age advanced. Maximum values for NDF, ADF, cellulose and lignin were observed in stem followed by whole plant and leaf, whereas hemicellulose, cutin and silica contents were higher in leaf fraction of the plant. The cultivars were observed to have some effects on chemical composition of all plant fraction. The results indicated that maturity had a much greater effect on the concentration of all the structural components than did the cultivars. It was concluded that maize fodder should be cut preferably between 8th to 9th week of age (flowering stage) to obtain more nutritious and digestible feed for livestock. Among the maize cultivars, Neelum proved to be the best, due to its higher dry matter contents and lower lignin concentration.

• Korean Journal of Environmental Agriculture
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• v.16 no.1
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• pp.55-60
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• 1997

Heavy metal-tolerant microorganisms, such as Pseudomonas putida, P. aeruginosa, P. chlororaphis and P. stutzeri which possessed the ability to accumulate cadmium, lead, zinc and copper, respectively, were isolated from industrial wastewaters and mine wastewaters polluted with various heavy metals. The distribution of heavy metal in the cell components, and amino acid compositions, was investigated. The distribution of heavy metal in the cell fractions of each heavy metal-tolerant microorganism grown for 20 hours in the basal medium containing 100mg/l of each heavy metal was investigated. In the case of cadmium-tolerant P. putida, lead-tolerant P. aeruginosa and copper-tolerant P. stutzeri, approximately $50{\sim}60%,\;30{\sim}40%$ and $10{\sim}17%$ of each heavy metal absorbed were distributed to cell wall, cell membrane and cytoplasm fractions, respectively. In the case of zinc-tolerant P. chlororaphis, approximately 32%, 55% and 13% of zinc were distributed to cell wall, cell membrane and cytoplasm fractions, respectively. These results indicated that the cell wall was a major adsorbing fraction of cadmium, lead and copper, and the cell membrane was that of zinc. Total amino acid content per gram of the cell grown in the culture media with heavy metal was higher than that of the cell grown in the culture media without heavy metal, and the content of acidic amino acids, such as aspartic acid(Asp.+Asn.) and glutamic acid(Glu.+Gln.) was higher than that of basic amino acids, such as histidine, lysine and arginine.

• Biotechnology and Bioprocess Engineering:BBE
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• v.11 no.1
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• pp.38-42
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• 2006

When whole cells are subjected to pyrolysis gas chromatography/mass spectrometry (Py-GC/MS) analysis, it provides biochemical profiles containing overlapping signals of the majority of compounds. To determine marker compounds that discriminate embryogenic calluses from nonembryogenic calluses, samples of embryogenic and nonembryogenic calluses of five higher plant species were subjected to Py-GC/MS. Genetic programming of Py-GC/MS data was able to discriminate embryogenic calluses from nonembryogenic calluses. The content ratio of 5-meyhyl-2-furancarboxaldehyde and 5-(hydroxymethyl)-2-furancarboxaldehyde was greater in nonembryogenic calluses than in embryogenic calluses. However, the content ratio of phenol, p-cresol, and $^1H-indole$ in embryogenic calluses was 1.2 to 2.4 times greater than the ratio in nonembryogenic calluses. These pyrolysates seem to be derived from the components of the cell walls, which suggests that differences in cell wall components or changes in the architecture of the cell wall playa crucial role in determining the embryogenic competence of calluses.

• Journal of the Korean Society of Food Culture
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• v.6 no.3
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• pp.229-235
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• 1991

This study is performed to investigate the relation between cell components and hardness which affect to the quality of dried persimmons. Moisture contents of dried persimmons were interrange of 30 to 36%. Enpung Junsi (EJ), Dungsi (Young dong) (DY) and Haman Susi (HS) had higher moisture contents than Dungsi (Sang Ju) (DS), Kojongsi (KS) and Hagakure (HK). Hardness was high in the varieties which has low moisture content such as DS, KS and HK. The significant differences of hardness and fracturability among varieties were appeared conspicuously but those of cohesiveness, adhesiveness and springiness were not appeared. Crude cell wall content, pectin and calcium content of cell wall were high in hard varieties. In the pectin fractions, water soluble pectin content was high in EJ, HS and DY but that of acid and alkali soluble pectin were high in DS, KS and HK. The cell wall of high hardness varieties observed thick and firm than that of low hardness varieties.

• Journal of Microbiology and Biotechnology
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• v.13 no.2
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• pp.202-206
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• 2003

To determine the effect of immunopotentiating activity of cellular components of Lactococcus lactis ssp. lactis, the immune function was analyzed in vitro using mice cells. When stimulated with mitogens, productions of $IFN-{\gamma}$, IL-12, $TNF-{\alpha}$, and IL-6 were enhanced in spleen cells treated with cellular components, with IL-4 production being the highest in spleen cells treated with cytoplasm fraction. Without mitogen stimulation, the productions of $IFN-{\gamma}$ and IL-12 were the highest in spleen cells treated with heat-killed whole cell. $TNF-{\alpha}$ and IL-6 productions were also high in spleen cells treated with all cellular components. Only heat-killed whole cell showed significant enhancement in natural killer cell activity. In peritoneal exudates cells, $TNF-{\alpha}$ production was enhanced significantly by all cellular components of Lactococcus lactis ssp. lactis These results indicate that the cellular components of Lactococcus lactis ssp. lactis are capable of stimulating immune cells to produce cytokines, and that both their cell walls and cytoplasm fraction contribute to these capacities.