• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1725-1731
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• 2002

The distribution and activities of hydrolytic enzymes (cellulolyti, hemicellulolytic,pectinolytic and others) in the rumen compartments of Hereford bulls fed 100% alfalfa hay based diets were evaluated. The alfalfa proportion in the diet was gradually increased for two weeks. Whole rumen contents were processed into four fractions: Rumen contents including both the liquid and solid fractions were homogenized and centrifuged, and the supernatant was assayed for enzymes located in whole rumen contents (WRE); rumen contents were centrifuged and the supernatant was assayed for enzymes located in rumen fluids (RFE); feed particles in rumen contents were separated manually, washed with buffer, resuspended in an equal volume of buffer, homogenized and centrifuged and supernatant was assayed for enzymes associated with feed particles (FAE); and rumen microbial cell fraction was separated by centrifugation, suspended in an equal volume of buffer, sonicated and centrifuged, and the supernatant was assayed for enzymes bound with microbial cells (CBE). It was found that polysaccharide-degrading proteins such as $\beta$-1,4-D-endoglucanase, $\beta$-1,4-D-exoglucanase, xylanase and pectinase enzymes were located mainly with the cell bound (CBE) fraction. However, $\beta$-D-glucosidase, $\beta$-D-fucosidase, acetylesterase, and $\alpha$-L-arabinofuranosidase were located in the rumen fluids (RFE) fraction. Protease activity distributions were 37.7, 22.1 and 40.2%, and amylase activity distributions were 51.6, 18.2 and 30.2% for the RFE, FAE and CBE fractions, respectively. These results indicated that protease is located mainly in rumen fluid and with microbial cells, whereas amylase was located mainly in the rumen fluid.

• Asian-Australasian Journal of Animal Sciences
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• 제33권12호
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• pp.1948-1956
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• 2020

Objective: The purpose of this study was to reveal the metabolic shift in the fungus cocultured with the methanogen (Methanobrevibacter thaueri). Methods: Gas chromatography-mass spectrometry was used to investigate the metabolites in anaerobic fungal (Pecoramyces sp. F1) cells and the supernatant. Results: A total of 104 and 102 metabolites were detected in the fungal cells and the supernatant, respectively. The partial least squares-discriminant analysis showed that the metabolite profiles in both the fungal cell and the supernatant were distinctly shifted when co-cultured with methanogen. Statistically, 16 and 30 metabolites were significantly (p<0.05) affected in the fungal cell and the supernatant, respectively by the co-cultured methanogen. Metabolic pathway analysis showed that co-culturing with methanogen reduced the production of lactate from pyruvate in the cytosol and increased metabolism in the hydrogenosomes of the anaerobic fungus. Citrate was accumulated in the cytosol of the fungus co-cultured with the methanogen. Conclusion: The co-culture of the anaerobic fungus and the methanogen is a good model for studying the microbial interaction between H₂-producing and H₂-utilizing microorganisms. However, metabolism in hydrogenosome needs to be further studied to gain better insight in the hydrogen transfer among microorganisms.

• KSBB Journal
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• 제18권5호
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• pp.371-376
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• 2003

Antioxidative and cholesterol-reducing activity of Bacillus polyfermenticus SCD were measured to characterize its probiotic properties. DPPH (1,1-diphenyl-2-picyryl hydrazyl) radical scavenging activity of the culture supernatant of B. polyfermenticus SCD was estimated to be 48%. The culture supernatant on the peroxidation of linoleic acid were investigated and the value was shown to be about 45%. The inhibition of TBA (2-thiobarbituric acid) formation of the culture supernatant was revealed 60% when stimulator was presented. The SOD-like activity of the culture supernatant was about 15%, which is similar to BHT (butylated hydroxytoluene) and ${\alpha}$-tocopherol. After cultured in TSB broth added soluble cholesterol either 0.1% or 0.3% of oxgall in 37$^{\circ}C$ for 24 h aerobically, cholesterol-reducing activities were revealed about 67% or 64%, respectively. To test whether the products are cholesterol-related or not, residual activity was determined. The cholesterol activity was rarely changed. In addition, when the cell extracts recovered after cultivation, was tested in absence of cholesterol, cholesterol activity was not detected. However, cholesterol activity was detected in the presence of cholesterol. Thus, it was assumed that B. polyfermenticus SCD could reduce cholesterol by conjugating with it, rather than by digesting the cholesterol using cholesterol-hydrolyzing enzymes.

• Korean Journal of Microbiology
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• 제44권2호
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• pp.140-146
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• 2008

Human cytomegalovirus (HCMV) stimulates the expression of intercellular adhesion molecule (ICAM-l) on the surface of monocytic THP-1 cells. Stimulation of ICAM-l did not require HCMV gene expression since UV-inactivated HCMV (UV-HCMV) was able to induce ICAM-l expression. ICAM-l expression was also stimulated in uninfected THP-l cells which were fed with culture supernatant of HCMV-infected THP-l cells. Co-culture experiment using trans-well insert supported that HCMV-infected THP-l cells secreted some cytokine(s) stimulating ICAM-l expression. The stimulation of ICAM-l by HCMV-infected cell culture supernatant appears to involve $NF-{\kappa}B$ pathway. Culture supernatant from THP-l cells infected with UV-HCMV, whose gene expression was abrogated, failed to stimulate ICAM-l expression on naive THP-l cells. Thus, HCMV gene expression seems to be required in secretion of cytokine(s) stimulating ICAM-l expression.

• Journal of Wetlands Research
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• 제16권3호
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• pp.347-353
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• 2014

This work was performed to find the effect of operation parameters on the permeate flux through the activated sludge dynamic layer, and to indicate the relationship between the water quality of supernatant and flux based on the results. Since the effluent can be obtained through steady and stable formation of cake layer in the bio-filter media system, it is an important subject to keep and control microbes with activated state in the bio-reactor. Filtration resistance was drastically increased at more than 18000mg/L of MLSS. With filtration time continued, the flux was gradually decreased and the water qualities of supernatant monitored by turbidity and TOC were also deteriorated. This phenomenon indicated that the organic materials generated by microbes and accumulated in the reactor might affect the flux in the system. In addition, the decrease of flux was simultaneously observed in the sludge volume index. When SVI was controlled from 150 to 250, the flux was also decreased. The proper aeration time was recommended to 30 to 60 seconds in this system. In order to operate this system steadily, therefore, the control of water quality of supernatant and SVI should be proceeded.

• Tuberculosis and Respiratory Diseases
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• 제42권4호
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• pp.447-454
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• 1995

Cytokine release from alveolar macrophages and subsequent interaction of these cytokines with the bronchial epithelium can induce epithelial cells to release inflammatory mediators. Nitric oxide(NO), a highly reactive gas formed from arginine by nitric oxide synthase(NOS), is known to be involved in inflammation and edema formation, and the inducible form of NOS(iNOS) can be increased by cytokines. In this context, we hypothesized that lung epithelial cells could be stimulated by cytokines released by alveolar macrophages to express iNOS. To test this hypothesis, the murine lung epithelial cell line, LA-4, or the human lung epithelial cell line, A549, were stimulated with culture supernatant fluids from alveolar macrophages. NO production was assessed by evaluating the culture supernatant fluids for nitrite and nitrate, the stable end products of NO. Both murine and human cell culture supernatant fluids demonstrated an increase in nitrite and nitrate which were time- and dose-dependent and attenuated by $TNF{\alpha}$ and IL-$1{\beta}$ antibodies(p<0.05, all comparisons). Consistent with these observations, cytomix a combination of $TNF{\alpha}$, IL-$1{\beta}$, and $\gamma$-interferon, stimulated the lung epithelial cell lines as well as primary cultures of human bronchial epithelial cells to increase their NO production as evidenced by an increase in nitrite and nitrate in their culture supernatant fluids, an increase in the iNOS staining by immunocytochemistry, and an increase in iNOS mRNA by Northern blottin(p<0.05, all comparisons). The cytokine effects on iNOS were all attenuated by dexamethasone. To determine if these in vitro observations are reflected in vivo, exhaled NO was measured and found to be increased in asthmatics not receiving corticosteroids. These data demonstrate that alveolar macrophage derived cytokines increase iNOS expression in lung epithelial cells and that these in vitro observations are mirrored by increased exhaled NO levels in asthmatics. Increased NO in the lung may contribute to edema formation and airway narrowing.

• Journal of the korean academy of Pediatric Dentistry
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• 제27권2호
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• pp.185-191
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• 2000

The water-soluble glucan produced by Lactococcus lactis 1370 affects the formation of dental plaque by Streptococcus mutans. In this study the factors affecting the formation of water-soluble glucan were assessed as the optical density of culture supernatant of Lactococcus lactis 1370 in the spectrophotometer. 1. The optical density of culture supernatant was high when Lactococcus lactis 1370 was cultured in M17 broth with 5% sucrose, while being low in culture supernatant of Streptococcus mutans. 2. The optical density of culture supernatant was higher when Lactococcus lactis 1370 was cultured in M17 broth with 10% sucrose than when being cultured without sucrose (p<0.05), and was higher at pH 7 than pH 5 (p<0.05). 3. The optical density of culture supernatant was the highest at $37^{\circ}C$ among $32^{\circ}C,\;37^{\circ}C\;and\;42^{\circ}C$, and was higher in the anaerobic incubator than in the aerobic incubator (p<0.05). 4. The optical density of culture supernatant was the highest in the media containing 1.0mM $CaCl_2$ (p<0.05), 2.5mM KCl (p<0.05), and 1.6mM $MgCl_2$. 5. When Streptococcus mutans was cultured in the media containing a quarter culture supernatant of Lactococcus lactis 1370 grown in M17 broth, the mean weight of produced artificial plaque was 103.0mg on the wire, whereas being significantly reduced to 5.6mg in the media containing a quarter culture supernatant of Lactococcus lactis 1370 grown in M17 broth containing 5% sucrose (p<0.05). These results indicate that the water-soluble glucan is more formed by Lactococcus lactis 1370 in the media containing sucrose or under the adequate conditions for the growth of bacteria, and inhibits the formation of artificial plaque by Streptococcus mutans.

• KSBB Journal
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• 제23권3호
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• pp.187-192
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• 2008

Lysozyme-producing microorganisms were isolated to obtain bacteria which can efficiently solubilize microbial cells. Cells of normal and chloroform-treated Escherichia coli and Micrococcus Iysodeikticus were used as model substrates to isolate lysozyme-producing microorganisms and investigate the efficiency of cell lysis. The culture supernatant of the isolate New1 (98% similarity of 16S rDNA sequence with Thermomonas haemolytica) showed different lytic characteristics for different substrates. Thermal treatment (autoclave) of substrate cells showed a significant effect on cell solubilization by culture supernatant of the New1. For autoclaved substrate cells, E. coli, M. Iysodeikticus and chloroform-treated E. coli were solubilized by 58.7%, 49.4% and 79.1%, respectively, in the culture supernatant of New1. The lytic activity of New1 was mainly caused by lysozyme produced by the isolate. It was also showed that New1 exhibited high protease activity and a little cellulase activity.

• Toxicological Research
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• 제14권3호
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• pp.301-306
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• 1998

This experiment was carrid out to know the effects of heating and serum on hydroxyl radicals in embryonic mouse forebrain (cerebrum) culture. The heating to mouse embryonic cerebrum cells in culture was done in a water bath at 43${\circ}C$ for 60min. After that, two supernatants were prepared at 20 hrs and 48 hrs respectively after heat treatment to the brain cells. To find out the heating effects on neuron cells, mouse cerebrum cells (13 embryonic day) were cultured in hydroxyl radical generation system composed of 20mU/ml glucose oxidase (GO system), using condition of normal culture media (MEM, 5% serum, 5% $CO_2$or supernatant prepared after heating at 43${\circ}C$ for 60 min in a water bath. Supernatant prepared at 20 hrs after heat treatment had a greater protective effects against hydroxyl radical than supernatant prepared at 48 hrs after heat treatment . Otherwise, the protective effect of serum against hydroxyl radicals in the cultured brain cells is higher than that in the heat treatment. These results indicated that serum in culture media reduced cytotoxicity of hydroxyl radicals in mouse forebrain culture, also that heat treatment showed the protective effects against hydroxyl radicals generated with 20mU/ml GO system in mouse forebrain culture.

• Journal of Veterinary Clinics
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• 제21권3호
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• pp.236-242
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• 2004

1,2-benzopyrone can stimulate macrophages to increase the ability of phagocytosis. Peripheral blood polymorphonuclear cells (PMN) and macrophages destroy microbial organisms with reactive oxygen species (ROS), called oxidative burst activity (OBA). This study was undertaken to determine whether 1,2-benzopyrone affects the OBA on the phagocytic response of canine peripheral blood phagocytes. The OBA of phagocytes in the addition or absence of latex beads was analyzed by flow cytometry system using dihydrorhodamine 123 (DHR). The direct treatments of 1,2-benzopyrone have no effect on the OBA of peripheral blood mononuclear cells (PBMC), PMN and monocyte-rich cells regardless of addition of latex beads. When latex beads are added to PMN, the OBA of PMN was remarkably enhanced by culture supernatant from PBMC but not PMN treated with 1,2-benzopyrone. Similary, it was also enhanced by human recombinant (hr) $TNF-\alpha.$ However, when latex beads were not added to PMN, its OBA was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. The OBA of latex beads-phagocytized PBMC and monocyte-rich cells was not enhanced by culture supernatant from either PBMC or PMN treated with 1,2-benzopyrone. These results strongly suggested that 1,2-benzopyrone has an immunoenhancing effect on the OBA of PMN when phagocytic response occurred only. This enhanced OBA may be mediated through active humoral substance(s), such as $TNF-\alpha,$ produced by PBMC stimulated with 1,2-benzopyrone.