Seo, Geon-Sik;Kim, Byung-Ryun;Park, Myeung-Soo;Kim, Min-Kyung;Yu, Seung-Hun
The Korean Journal of Mycology
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v.30
no.2
/
pp.86-94
/
2002
Recently a serious outbreak of weed mould caused by a species of Hypocrea occurred in oyster mushroom (Pleurotus ostreatus) substrates in Korea. The disease was characterized by a rapid infestation of the oyster mushroom substrates by Hypocrea sp. and subsequent inhibition of fructification of the mushroom. In spite of it's serious losses to the oyster mushroom industry in Korea, etiology and ecology of the disease have not been studied. Morphological characteristics of the fungus were examined and molecular characteristics of the fungus were compared with those of the green moulds (Trichoderma spp.) isolated from oyster mushroom bed. Stromata formed superficially on suface of the substrates were pulvinate to effuse or irreguler, initially white but becoming yellowish brown, measuring $6.0{\sim}13.0{\times}3.0{\sim}11.0mm$. Perithecia were globose to subglobose, immersed in stroma, $223{\sim}263\;(Ave.239.9){\times}167.3{\sim}231\;(Ave.204.1){\mu}m$ in size. Asci were unitunicate, cylindrical, nonamyloid, $82.7{\sim}124.8\;(Ave.103.3){\times}4.1{\sim}5.1\;(Ave.4.9){\mu}m$ in size, 16 part-spored. Ascospores were bullet-shaped or somewhat oblong, hyaline, bicellular, roughened or warted, $5.4{\sim}7.4\;(Ave.6.5){\times}3.6{\sim}5.5\;(Ave.4.7){\mu}m$ in size. This fungus readily form the stroma on PDA. Mycelia on PDA nearly invisible and without cottony aerial mycelium. Optimum temperature for mycelial growth of this fungus was $25^{\circ}C$ on PDA and its growth rate was 15 mm per day. This species did not grow at below 10 and above $35^{\circ}C$. Phialides in culture enlarged in the middle and aggregated to penicillate type. They were very variable, shorted ampulliform and occasionally curved when matured, but cylinderical when young, measuring $11.9{\sim}24.3\;(Ave.\;14.7){\times}2.9{\sim}3.9\;(Ave.\;3.4){\mu}m$ when matured and $7.2{\sim}14.0\;(Ave.\;10.8){\times}2.8{\sim}4.9\;(Ave.\;3.5){\mu}m$ when young. Phialosopres were ovoid to ellipsoid, smooth, measuring $3.5{\sim}7.2\;(Ave.\;4.5){\times}2.6{\sim}3.3\;(Ave.\;2.9){\mu}m$. Nineteen isolates of Hypocrea sp. were analyzed on the basis of molecular characteristics and classified into phenotypic groups. On the basis of RAPD, URP-PCR, the fungus was confirm to monoclonal, and was classified as a different taxon from reported species of Hypocrea and Trichoderma and supposed to be a new species not previously reported in literature.
A study was conducted to determine whether the vaccination programs for the control of New castle disease (ND) would affect the immune status of birds against the disease. Twenty-six poultry flocks in the epizootic area of ND were surveyed to investigate the level of urn antibody against ND virus and the programs used for the vaccination of birds. The mortality rates and vaccination status of birds during the epizootic of ND were also examined in the infected poultry flocks to elucidate the immune effect against the epizootic with particular regard to various vaccination programs used in the field. The results obtained are summerized as follows: 1. Of 26 poultry flocks investigated, 22 flocks were immunized with live and killed vaccines, their haemagglutination-inhibition (HI) antibody titer being 146 and 50, respectively. Among 22 farms using live and killed vaccines two flocks which showed the lowest HI titer of 10 and 23 had the disease later on. However, no cases of ND were recorded in the killed vaccine groups, although their HI titers were in the range of 38 to 64. 2. Of 14 infected flecks, one flock was not vaccmated against ND while all the remaining 13 flocks were vaccinated against the disease, of which 8 flocks were vaccinated with live vaccine only and the other 5 flocks with both live and killed vaccines. The mortality rate of 8 infected flocks which had been vaccinated with only live vaccine was as high as 32.5% while that of 5 flocks with both live and killed vaccines was as low as 5.1%. 3. It was found that in majority of flocks B$_1$vaccine was used via drinking water and in a few flocks the vaccine was administered via intramuscular route or method of dipping mouth, nose and eye of birds into vaccine solution.
The experiment using Anake broiler strain chicks was conducted to study the effort fungistatic agents on microbial counts, Ins of nutrient, growth rate and feed efficiency of the broiler. Feed was adjusted to 12% and 15% moisture level during hot and high humidity season and sorbic acid at the level of 0.02%, 0.04% and Ca-propionic acid at the level 0.1% 0.2% was added. The results obtained were as follows. 1, The Addition of fungistatic agents could slightly reduce mold and yeast counts. The highest effect on inhibition of mold and yeast counts was observed for the addition of sorbic acid at 0.04% level to the fled contained 15% moisture. 2, Approximately 14% starch loss of ground corn was observed from the fred contained 15% moisture and the loss could be diminished to 3-7% by the addition of fungistatic agents with the superior effect of sorbic acid to Ca-propionic acid. 3. Approximately 15% fat loss was detected when high moisture fled was und and this was reduced to 7% by the addition of 0.04% sorbic acid to the feed. 4. Significantly higher growth rate (p<0.05) during starter period was observed for low moisture feed added by sorbic acid compared with that for high moisture diet without fungistatic agents or with Ca-propionate at the level of 0.1%. 5. Significantly lower feed efficiency (p<0.05) during starter period of high moisture feed without fungistatic agents was observed; hower no significant different response was detected by either moisturer level of kinds and levels of fungistatic agents used. 6. Significantly higher growth rate (p<0.05) during finisher period of lower moisture feed with 0.04% sorbic acid was observed compared with that of high moisture fled without fungistatic agents or the feed added by 0.1% Ca-propionate level. 7. Significantly higher feed efficiency (p<0.05) during finisher period of low moisture feed added by sorbic acid 0.02% or 0.04% level was found compared with that of high moisture feed without fungistatic agents and low moisture feed added by Ca-propionate at the level of 0.1% or 0.2%. 8. Mort bacteria, mold and yeast were observed in the ceca than in small intestine. The. moisture content of dict had no effect on intestinal microfloral counts. However, numbers of mold and yeast of intestine could slightly be reduced by fungistatic agents administration. 9. Nothing but encephalomalacia to chicks fed feed contained 15% moisture without addition of fungistatic agents was observed. In conclusion, addition of either sorbid acid at 0.04% level and Ca-propionate at 0.2% level to high moisture feed or reduced moisture level to 12% could be con-sidered more effective to enhance growth rate and fled efficiency of broilers during summer period.
This study was conducted to investigate the effects of bifidobacteria on the growth and Caco-2 cell-adherence of Escherichia coli O157:H7 .Dur-ing momo-culture of E. coli O157:H7 and mixed culture with Bifidobacterium infantis K9, pH viable cell count, and ammonia concentration were measured Co-cultivation of E. coli O157:H7 with bifidobacteria. producing acidic metabolites rapidly decreased the viable cell count of E. coli O157:H7 In addition rapid decrease of ammo- nia concentration was observed during mixed culture after 8 hrs incubation compared to single culture of E. coli O157:H7 Therefore it is likely that bifidobacteria assimilate ammonia produced by E. coli O157:H7 P4 B, infantis K9 showed quite similar adherence on the Caco-2 cells in either case. On the other hand adherence of E. coli O157:H7 decreased from 2.6% to 1.86% when B infantis K9 was adhered to Caco-2 cell 2 hrs prior to the application of E. coli O157:H7 In conclusion in adherence of E coli O157:H7 to Caco -2 cell was inhibited by competition of its binding to the adherence site with bifidobacteria. In addition inhibitory effects of bifidobacteria on E coli O157:H7 appeared to be much higher with increae of the number of bifidobacteria and its ability of adherence to Caco-2 cells.
This study was carried out to investigate the effects of phenobarbital sodium(PB) and 3-methylcholanthrene(3-MC) on carbofuran cytotoxicity and to develop antitoxic agents based on the effectivness. Experimental groups for carbofuran cytotoxicity were divided into five groups ; medium alone and four treatments of carbofuran (1, 25, 50 and $100{\mu}M)$, and those for compensation effects were divided into six groups ; medium alone, $IC_{50}$ carbofuran and four combinations of carbofuran and PB or 3-MC($IC_{50}$ carbofuran plus 1, 25, 50, $100{\mu}M$ of PB and 3-MC, respectively). After incubation for 48 hrs under the same conditions, MTT(Tetrazolium MTT), NR(Neutral red) and SRB(Sulforhodamine B protein) assay were performed. Fifty percentage inhibition of MTT, NR, and SRB against carbofuran in rat fibroblast cell were 60.7, 82.5 and $87.0{\mu}M$, respectively. At the combination treatments of $IC_{50}$ of carbofuran and $100{\mu}M$ of PB, the significant compensation effects were observed from the results of MTT and NR but not from that of SRB absorbance. And at the combination treatments of $IC_{50}$ of carbofuran and 3-MC, the relatively significant compensation effects were found at $50{\mu}M$ 3-MC from the results of MTT and at $100{\mu}M$ 3-MC from that of NR and SRB absorbances, respectively. From the results of light microscopy, combination treatments of $carbofuran(IC_{50})$ and PB or 3-MC showed good regeneration in carbofuran toxicity of rat fibroblast cells. These results suggest that PB or 3-MC can compensate the cytotoxity of carbofuran insecticide in rat NIH3T3 fibroblast cells.
Park, Jae-Yong;Kim, Jeong-Ran;Chang, Hee-Jin;Kim, Chang-Ho;Park, Jae-Ho;Jung, Tae-Hoon
Tuberculosis and Respiratory Diseases
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v.45
no.3
/
pp.587-595
/
1998
Background: Recombinant adenovirus hold promise as vectors to carry therapeutic genes for several reasons: 1) they can infect both dividing and non-dividing cells; 2) they have the ability to directly transduce tissues in vivo; 3) they can easily be produced in high titer; and 4) they have an established record of safety as vaccination material. However, one of the major limitation in the use of adenoviruses is that transgene expression is quite short because adenovirusees insert their DNA genome episomally rather than by chromosomal integration, and an immune response against the virus destroys cells expressing the therapeutic gene. Since sodium butyrate has been reported to induce adenovirus-mediated gene expression, we hypothesized that treatment of tumor cells, transduced with herpes simples virus thymidine kinase(HSVtk) gene using adenoviral vector, with butyrate could augment the effect of gene therapy. Methods: We transduced HSVtk gene, driven by the cytomegalovirus promoter, into REN cell line(human mesothelioma cell line). Before proceeding with the comparison of HSVtk/ganciclovir mediated bystander killing, we evaluated the effect of butyrate on the growth of tumor cells in order to rule out a potential antitumor effect of butyrate alone, and also on expression of HSVtk gene by Western blot analysis. Then we determined the effects of butyrate on bystander-mediated cell killing in vitro. Results: There was no inhibition of growth of cells exposed to butyrate for 24 hours at a concentration of 1.5mM/L. Toxic effects were seen when the concentration of butyrate was greater than 2.0mM/L. Gene expression was more stable and bystander effect was augmented by butyrate treatment of a concentration of 1.5mM/L. Conclusion: These results provide evidence that butyrate can augment the efficiency of cell killing with HSVtk/GCV system by inducing transgene expression and may thus by a promising new approach to improve responses in gene therapy using adenoviral vectors.
Kim, Myong-Jo;Kim, Ju-Sung;Jeong, Dong-Myong;Ham, Seung-Shi;Yu, Chang-Yeon
Korean Journal of Medicinal Crop Science
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v.10
no.3
/
pp.222-229
/
2002
Ixeris dentata root were extracted with methanol and then fractionated with n-hexane, EtOAc and BuOH to get active fractions. and their antioxidant and antimicrobial activities in each fraction were determined. Ethyl acetate fraction of Ixeris dentata root showed strong antioxidant activities, but aqueous fraction did not show any activities. But in the antimicrobial test, aqueous fraction showed strong antimicrobial activities except to Escherichia coli. especially, aqueous fraction showed the strongest activities against Hypocrea nigricans. and butanol fraction showed the strongest activities against Cladosporium herbarum. This study was performed to determine the antimutagenic and cytotoxic effect of Ixeris dentata root methanol extract on Salmonella typhimurium TA98, TA100 and cancer cell lines using ames test and cytotoxicity assay, respectively. Cancer cell lines include human lung carcinoma(A549), human breast adenocarcinoma(MCF-7) and human hepatocellular carcinoma (Hep 3B). Futher fractionations with hexane, ethyl acetate, butanol and water from methanol extract of Ixeris dentata root were performed to obtain effective fraction, methanol extracts showed 79.94% inhibitory effect on the mutagenesis induced by N' -methyl- N' -nitro-N-nitrosoguanidine(MNNG) against TA100, while 89.99% inhibition was observed on the mutagenesis induced by 4-nitroquinoline-1-oxide(4NQO) against TA98. In the meanwhile, butanol fraction showed 89.92% and 71.01% inhibitory effect on the mutagenesis induced by benzo(a)pyrene(B(a)P) against TA98 and TA100, respectively. Ethyl acetate fraction showed the strongest effect against A549, MCF-7 and Hep3B at the same concentration compared to those of other fration.
In order to establish ELISA method to detect antibody against IBV various factors involved were examined. Antigen was prepared from Massachusetts type IBV which is known to be one of serotypes distributed most widely. The virus was grown in embryonated SPF chicken eggs. Allantoic fluid harvested was processed to ultracentrifugation and sucrose density gradient centrifugation to produce a purified antigen The antisera selected from the field samples based on hemagglutination inhibition test were used as the standard positive and negative sera for this study and the results obtained were summarized as follows. 1 , It was found that ELISA test was satisfactory when the purified antigen was coated on the plate in the amount of about 40ng protein per well. In case of the phospholipase treated hemagglutinating antigen it gave satisfactory results when the each well wns coated with 1.2 to 2.5 hemagglutinating unit which was equivalent to 40 to 90ng of protein. 2. There was no significant difference in the ratio of optical density of positive to that of negative serum whether the coated antigen was held for 1 hour at 37$^{\circ}C$ or it was held overnight at 4$^{\circ}C$. The coated antigen could be kept in dried state without change of antigenecity for at least one month of experimental period at 4$^{\circ}C$. 3. There was a big variation in the optical density and P/N values depending on the maker of the plates and on the plate of the same maker. 4. It was found that background optical density was negligible when serum was diluted more than 1:50 and serum dilution of 1:100 appeared to be appropriate as a routine test dilution to screen the antibody. 5. Optical density was fairly constant 15 minutes afterward from the time substrate was treated and during the 4 hours after stopper was treated. 6. There was a low correlation(r=0.42) between ELISA and HI test. However, when 74serum samples were tested for the IBV antibody, 98.7% were found to be positive by both tests in which titers of 2$^{6}$ or more by HI test and P/N values of 1.4 or more by ELISA were considered to be positive, 7 Day-old IBV vaccinated chickens shows a similar antibody decay and rising pattern until 8 weeks of age by the two tests, ELISA and HI.
Purpose: Poncirus trifoliata has been reported to have anti-inflammatory, antioxidant, and immune activities. However, its anti-obesity activity and the mechanism by which the water extract of dried, immature fruit of Poncirus trifoliata (PF-W) acts are not clear. This study suggests a potential mechanism associated with the anti-obesity activity of PF-W. Methods: We measured the effect of PF-W on lipoprotein lipase (LPL) regulation using enzyme-linked immunosorbent assay (ELISA) and an activity assay. The LPL regulation mechanism was examined by reverse transcription polymerase chain reaction (RT-PCR) to measure the mRNA expression of biomarkers related to protein transport and by western blot for analysis of the protein expression of the transcription factor CCAAT-enhancer-binding protein ($C/EBP{\beta}$). Results: The total polyphenol and flavonoid content of PF-W was $52.15{\pm}4.02$ and $6.56{\pm}0.47mg/g$, respectively. PF-W treatment decreased LPL content in media to $58{\pm}5%$ of that in control adipocyte media, and increased LPL content to $117{\pm}3.5%$ of that in control adipocytes, but did not affect the mRNA expression of LPL. PF-W also increased the mRNA expression of sortilin-related receptor (SorLA), a receptor that induces endocytosis and intracellular trafficking of LPL, in a concentration- and time-dependent manner. Finally, cell fractionation revealed that PF-W treatment induced the expression of $C/EBP{\beta}$, a SorLA transcription factor, in the nuclei of 3T3-L1 adipocytes. Conclusion: The LPL secretion and activity assay showed PF-W to be an LPL secretion inhibitor, and these results suggest the potential mechanism of PF-W involving inhibition of LPL secretion through $C/EBP{\beta}$-mediated induction of SorLA expression.
Kim, Ji-Eun;Joo, Sung-Il;Seo, Ji-Hyun;Lee, Sam-Pin
Journal of the Korean Society of Food Science and Nutrition
/
v.38
no.8
/
pp.989-995
/
2009
Extract yield of tartary buckwheat treated with water, 70% ethanol or methanol were about 13.6%, 7.0% and 6.6%, respectively. Extract yield was greatly increased by the treatment of $\alpha$-amylase indicating 95.1% yield. $RC_{50}$ value of DPPH radical scavenging activity with methanol and 70% ethanol extracts were 34.0 $\mu g$/mL, 40.5 $\mu g$/mL, respectively. The DPPH radical scavenging activity increased when it was treated with $\beta$-glucosidase and cellulase, showing $RC_{50}$ value of 24.7 $\mu g$/mL and 25.0 $\mu g$/mL, respectively. In ABTS radical scavenging activity, methanol extract (100 $\mu g$/mL) showed 30% inhibition. In DPPH or ABTS radical scavenging activities, the treatment of $\beta$-glucanase and $\alpha$-amylase shows the highest and the lowest activities, respectively. In $\alpha$-glucosidase inhibitory effect, 70% ethanol extract showed $RC_{50}$ value of 59.9 $\mu g$/mL, but water extract was not inhibitory effective. The $\alpha$-glucosidase inhibitory effect was the highest in multi enzyme treatment. Content of rutin and quercetin in methanol extract showed higher value with 4400.3 mg% and 71.9 mg%, respectively. The 70% ethanol extract of buckwheat contained rutin of 3459.8 mg% and quercetin of 56.9 mg%. In the treatment of $\beta$-glucanase, the rutin content of ethanol extract increased with 5057.4 mg% and multi-enzyme treatment resulted in the modification of rutin glycoside.
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