• Asian-Australasian Journal of Animal Sciences
• /
• v.16 no.4
• /
• pp.541-548
• /
• 2003

The Immunoglobulin $IgG_1$ and $IgG_2$ isotype immune responses of domestic ruminants and mice to Cowdria. ruminantium live infection or by immunization with inactivated organisms were determined by the enzyme linked immunosorbent assay and Western blotting. Immunization of goats with inactivated elementary bodies (IEBs) led to a predominant $IgG_1$ isotype response. This indicated that a Th2 response was induced. After challenge, the IgG isotype responses were mixed whereby both $IgG_1$ and $IgG_2$ antibodies were detected. Two goats that survived virulent challenge had a predominant $IgG_2$ isotype response. In cattle live infection by natur l challenge or experiment led to a predominant $IgG_1$ isotype response. Immunization of cattle with IEBs however led to mixed IgG responses characterized by similar $IgG_1$ and $IgG_2$ ratios. In the mouse live infection led to a predominant $IgG_2$ isotype response. This indicated the mouse developed a true Th1 type cell mediated immune response when inoculated with live organisms. Immunization with inactivated organisms on the other hand led to a dominant $IgG_1$ response. It is evident from this work that the immune responses of ruminants and mice to C. ruminantium are different and that using mice as the experimental model for immune responses to Cowdria ruminantium. is not the appropriate.

• Asian-Australasian Journal of Animal Sciences
• /
• v.30 no.6
• /
• pp.788-796
• /
• 2017

Objective: In order to improve fermentation quality of natural grasses, their silage preparation and fermentation quality in meadow steppe (MS) and typical steppe (TS) were studied. Methods: The small-scale silages and round bale silages of mixed natural grasses in both steppes were prepared using the commercial lactic acid bacteria (LAB) inoculants Chikuso-1 (CH, Lactobacillus plantarum) and cellulase enzyme (AC, Acremonium cellulase) as additives. Results: MS and TS contained 33 and 9 species of natural grasses, respectively. Stipa baicalensis in MS and Stipa grandi in TS were the dominant grasses with the highest dry matter (DM) yield. The crude protein (CP), neutral detergent fiber and water-soluble carbohydrate of the mixed natural grasses in both steppes were 8.02% to 9.03%, 66.75% to 69.47%, and 2.02% to 2.20% on a DM basis, respectively. All silages treated with LAB and cellulase were well preserved with lower pH, butyric acid and ammonia-N content, and higher lactic acid and CP content than those of control in four kinds of silages. Compared with CH- or AC-treated silages, the CH+ AC-treated silages had higher lactic acid content. Conclusion: The results confirmed that combination with LAB and cellulase may result in beneficial effects by improving the natural grass silage fermentation in both grasslands.

• The Journal of the Korean Society for Microbiology
• /
• v.34 no.6
• /
• pp.573-582
• /
• 1999

Cytokines are hormone-like proteins which mediate and regulate inflammatory and immune responses. The purpose of this study was to investigate the effect of lipopolysaccharide (LPS), Staphylococcus enterotoxin B (SEB), and Mycoplasma lysates on regulation of IL-6 and IL-8 production by human nasal fibroblasts. Primary cultured cells were incubated with LPS ($1.0\;{\mu}g/ml$) from E.coli, SEB ($1.0\;{\mu}g/ml$) from S.aureus, or Mycoplasma lysates (M.pneumoniae, Mp; M. fermentans, Mf; M. hominis, Mh, each $1.0\;{\mu}g/ml$). The culture supernatants were collected at 2, 6, and 24 hr and assessed for IL-6 and IL-8 production by enzyme-linked immunosorbent assay. The production of IL-6 in the culture supernatant was downregulated by LPS, SEB, or Mycoplasma lysates. But IL-6 was upregulated by mixed exposure with Mp+LPS (2 hr), Mp+LPS+SEB (24 hr), Mf+LPS (24 hr), Mf+LPS+SEB (2 hr), Mh+LPS (24 hr), Mh+SEB (24 hr), or Mh+LPS+SEB (24 hr). The production of IL-8 in the culture supernatant was similar to that of IL-6 by same stimulants. But IL-8 was upregulated by mixed exposure with Mf+LPS+SEB (2 hr), Mh+LPS (24 hr), Mh+ SEB (24 hr), or Mh+LPS+SEB (24 hr). These studies show that costimulation of LPS or SEB with Mycoplasma whole cell lysates upregulates the production of IL-6 and IL-8.

• Journal of the Korean Wood Science and Technology
• /
• v.17 no.3
• /
• pp.52-60
• /
• 1989

This study was carried out to know the possibility of simultaneous utilization of laccase from white-rot fungus with cellulase on enzymatic hydrolysis of cellulosic substrate from autohydrolyzed oak wood. Laccases from 3 white-rot fungi, Pleurotus ostreatus. Ganoderma lucidum, and Phanerochaete chrysosporium, were isolated, purified and measured their activities. The highest activity was shown in Pleurotus ostreatus and the lowest in Phanerochaete chrysosporium. Laccase from Pleurotus ostreatus has optimum pH of 5.94, Km value of 3.209 mM and appeared to be stable at relatively wide pH range, 4.7-8.72. Temperature stability showed that 60% activity was preserved after 40 minutes at $50^{\circ}C$. Laccase from Ganoderma lucidum reached to the maximum activity during 15-20 day incubation. This enzyme has optimum pH of 6.45, Km value of 6.71 mM and pH range of 5.0-9.0 for stabilization. 95% activity was preserved at $30^{\circ}C$ and 58% activity at $50^{\circ}C$. Concerned to the enzymatic hydrolysis of cellulosic substrate with both enzymes, cellulase and laccase, simultaneously, mixed culture filtrates and mycellium extracts were shown higher hydrolysis rates than those of Trichoderma viride. There were no significant differences in the extent of hydrolysis among various mixed culture filtrates and mycellium extracts.

• Microbiology and Biotechnology Letters
• /
• v.31 no.3
• /
• pp.257-263
• /
• 2003

The cellulases production in solid state fermentation (SSF) of Trichoderma harzianum FJ1 with high cellulases productivity using cellulosic wastes was investigated. Physical and chemical conditions of the fermentation, such as moisture content, initial pH, and composition of mixed substrate (wine waste, rice straw, and soybean flour) on FPase (Filter paper activity) production were examined. The enzyme production was optimized in the conditions of moisture content of 70%, pH 5.0, 3$0^{\circ}C$, and 1:1:1 composition of mixed substrate containing wine waste, rice straw, and soybean flour. The highest activities of FPA, CMCase, Xylanase, $\beta$-glucosidase, and Avicelase in the optimized culture conditions were 15.2, 69.1, 83.9, 29.2, and 4.2 unit/g-SDW in 5 day cultivation, respectively. Economical and efficient production of cellulolytic enzymes by T harzianum FJ1 using cellulosic wastes in solid state fermentation will contribute to the biological saccharification of cellulosic wastes with enormous potential resource value in future.

• Journal of the Korea Organic Resources Recycling Association
• /
• v.9 no.4
• /
• pp.99-104
• /
• 2001

The influence of agitation speed on the yeast growth was investigated in the production of probiotic feed from pulverized liquified food wastes by aerobic fermentation. A yeast Kluyvermyces marxianus was selected through a preliminary screening. The yeast was cultured by 2liter jar fermenter. in 10% solid(w/v) substrate of liquified food waste at $35^{\circ}C$ with each different agitation speed of 500, 900 and 1200 rpm. For the acceleration of enzyme excretion mixed culture with Aspergillus oryzae was also attempted and the results were compared to those of single culture. As results the viable cell number was increased by increasing agitation speed. But it showed highest value in 900rpm and then decreased in 1200rpm. The mixed culture increased amylase activity and growth rate, but did not seem to enhance the highest viable cell count in the final fermentation stage.

• Environmental Analysis Health and Toxicology
• /
• v.21 no.2 s.53
• /
• pp.127-138
• /
• 2006

In urban areas, diesel exhaust particles (DEP) are probably a major component of particulate matters, especially in Korea where drive many diesel vehicles. The aim of this study was to investigate genotoxic effects of DEP using single ceil gel electrophoresis. In order to evaluate the mechanisms of DEP genotoxicity, the rat microsome mediated and DNA repair enzyme treated comet assays together with conventional comet assay were performed. Total diesel particles (DEPT) was collected without site fractionation from diesel engine bus and dichloromethane extract was obtained. The organic extract of DEPT revealed DNA damage itself in NIH/3T3 cells. The level of DNA breaks plus oxidative DNA lesions and microsome mediated DNA damage was assessed by modified single cell gel eletrophoresis. DEPT was able to induce oxidative DNA damage as well as microsome mediated DNA damage. Vitamin C as an model antioxidant reduced DNA damage in endonuclase III treated comet assay. One of flavonoid, galangin as a CYP1A1 inhibitor. reduced DNA damage in the presence of S-9 mixture. $DEP_T$ is the sources of oxidative stress, but antioxidants can significantly reduce oxidative DNA dmage. And $DEP_T$ may contain indirect mutagens which can be inhibited by CYP1A1 inhibitors. The ethanol extracts of the mixed vegetables (BV) or the mixed fruits (BF) were evaluated for their in vitro antigenotoxic effects. BV and BF showed potent Inhibitory effects against DEPT induced DNA damage with oxidative DNA lesions and in the prescence of S-9 mixture. These results indicate that BV and BF could prevent cellular DNA damage by inhibiting oxidative stress and suppressing cytochrome P4501A1 in cell culture.

• Asian-Australasian Journal of Animal Sciences
• /
• v.19 no.1
• /
• pp.137-143
• /
• 2006

The purpose of this study was to prepare a binder containing porcine blood transglutaminase (TGase), thrombin and fibrinogen. Extracted TGase, thrombin and fibrinogen were used alone or mixed with different proportions of their volume (v/v/v) by nine combinations as follows were 0.5:1:15, 0.5:1:20, 0.5:1:25, 1:1:15, 1:1:20, 1:1:25, 1.5:1:15, 1.5:1:20 and 1.5:1:25, respectively. Five ml of each combination were mixed with 0.6 ml of 0.25 M calcium chloride before experiment. After storage at 4C for 0, 1, 2, 3, 4 and 5 weeks, enzyme activity, total plate count, pH value, and SDS-PAGE of TGase, thrombin and fibrinogen were tested and pH value, clotting time and gel strength of the nine combination binders were determined. The results showed that total plate count of thrombin and pH value of TGase were significantly higher (p<0.05) than in other treatments. SDS-PAGE results showed that purified TGase, thrombin and fibrinogen from porcine blood plasma compared with commercial products (Sigma) had the same band patterns and nine different combination binders had no significant effect. Enzymatic activity of TGase and thrombin decreased as storage time increased. Total plate count of TGase, thrombin and fibrinogen and clotting time of the binder increased as storage time increased. The higher amount of fibrinogen in combinations, the stronger the gel strength.

• Restorative Dentistry and Endodontics
• /
• v.19 no.2
• /
• pp.409-428
• /
• 1994

Endodontic surgery is performed when conventional endodontic therapy fails or is contraindicated. In such cases, retrograde filling materials including amalgam, composite resin, and various cements have been used. Biocompatibilty and margin sealing ability of retrograde filling materials are important for the long term success of endodontic surgery. In vitro cell culture is frequently used as the method of measuring the biocompatibilty of dental materials. The purpose of this study was to evaluate the cytotoxicity of six kinds of retrograde filling materials including newly developed light curing glass ionomer cements. Each material was mixed according to. the manufacture's instruction and evaluated as : freshly mixed, 24-hour after mixing, and 168-hour after mixing respectively. The elution solution was extracted after 24-hour contact with materials using media. Cytotoxicity was evaluated by direct contact, or elution contact. Test results of radiochromium($^{51}Cr$) release, cell viability using tetrazolium dye (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl dimethyltetrazolium bromide(MTT) test and lactate dehydrogenase(LD) of damaged L929 cells were analyzed. In the $^{51}Cr$ release of direct contact, all experimental retrograde filling materials except amalgam and glass ionomer cement showed increased cytotoxicity compared to control. In the $^{51}Cr$ release of elution solution, the released $^{51}Cr$ was so minimal that it was impossible. to evlauate the cytotoxicity exactly. The elution solutions of glass ionomer cement and IRM showed marked cytotoxicity in MTT test. LD enzyme activity was highest in tests of direct contact with composite, light curing composite, and light curing glass ionomer cement and IRM. Amalgam revealed least cytotoxicity while IRM showed cytotoxicity using all three methods. Composite, light curing composite and light curing glass iomomer cement were cytotoxic in the tests of $^{51}Cr$ release and LD activity. Glass ionomer cement showed cytotoxic effect only in the MTT method. From these results it is suggested that the standardization and optimization of cytotoxicity testing, especially using elution solutions, should be strongly advised.

• Korean Journal of Environmental Agriculture
• /
• v.13 no.1
• /
• pp.76-82
• /
• 1994

A competitive indirect enzyme-linked immunosorbent assay(ELISA) was developed to detect and quantify levels of the fungicide metalaxyl in crops. Antiserum against metalaxyl was demonstrated in rabbits immunized with metalaxyl-human serum albumin(HSA) conjugate. Metalaxyl-protein conjugate was prepared by mixed anhydride and peptide coupling method with EDC. In this assay, metalaxyl-ovalbumin(OA) was $coated(8{\mu}g/ml)$ on the microtiter plate, which was incubated for 1 hr at $4^{\circ}C$ or 4 hr at $37^{\circ}C$ with diluted antiserum(1:2,000). The optimum volume ratio of antigen and antibody mixture was 0.5: 1, which was incubated for 1 hr at $20^{\circ}C$. The detection of metalaxyl bound on the surface of wells was determined by the reaction(30 min) of antirabbit Ig G-peroxidase conjugate with its substrate.