• Asian-Australasian Journal of Animal Sciences
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• v.24 no.2
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• pp.285-294
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• 2011

The abatement of methane emission from ruminants is an important global issue due to its contribution to greenhouse gas with carbon dioxide. Methane is generated in the rumen by methanogens (archaea) that utilize metabolic hydrogen ($H_2$) to reduce carbon dioxide, and is a significant electron sink in the rumen ecosystem. Therefore, the competition for hydrogen used for methanogenesis with alternative reductions of rumen microbes should be an effective option to reduce rumen methanogenesis. Some methanogens parasitically survive on the surface of ciliate protozoa, so that defaunation or decrease in protozoa number might contribute to abate methanogenesis. The most important issue for mitigation of rumen methanogenesis with manipulators is to secure safety for animals and their products and the environment. In this respect, prophylactic effects of probiotics, prebiotics and miscellaneous compounds to mitigate rumen methanogenesis have been developed instead of antibiotics, ionophores such as monensin, and lasalocid in Japan. Nitrate suppresses rumen methanogenesis by its reducing reaction in the rumen. However, excess intake of nitrate causes intoxication due to nitrite accumulation, which induces methemoglobinemia. The nitrite accumulation is attributed to a relatively higher rate of nitrate reduction to nitrite than nitrite to ammonia via nitroxyl and hydroxylamine. The in vitro and in vivo trials have been conducted to clarify the prophylactic effects of L-cysteine, some strains of lactic acid bacteria and yeast and/or ${\beta}$1-4 galactooligosaccharide on nitrate-nitrite intoxication and methanogenesis. The administration of nitrate with ${\beta}$1-4 galacto-oligosaccharide, Candida kefyr, and Lactococcus lactis subsp. lactis were suggested to possibly control rumen methanogenesis and prevent nitrite formation in the rumen. For prebiotics, nisin which is a bacteriocin produced by Lactococcus lactis subsp. lactis has been demonstrated to abate rumen methanogenesis in the same manner as monensin. A protein resistant anti-microbe (PRA) has been isolated from Lactobacillus plantarum as a manipulator to mitigate rumen methanogenesis. Recently, hydrogen peroxide was identified as a part of the manipulating effect of PRA on rumen methanogenesis. The suppressing effects of secondary metabolites from plants such as saponin and tannin on rumen methanogenesis have been examined. Especially, yucca schidigera extract, sarsaponin (steroidal glycosides), can suppress rumen methanogenesis thereby improving protein utilization efficiency. The cashew nutshell liquid (CNSL), or cashew shell oil, which is a natural resin found in the honeycomb structure of the cashew nutshell has been found to mitigate rumen methanogenesis. In an attempt to seek manipulators in the section on methane belching from ruminants, the arrangement of an inventory of mitigation technologies available for the Clean Development Mechanism (CDM) and Joint Implementation (JI) in the Kyoto mechanism has been advancing to target ruminant livestock in Asian and Pacific regions.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.3
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• pp.349-356
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• 2014

The objective of this study was to evaluate the effects of molasses and propionic acid on the fermentation quality and aerobic stability of total mixed ration (TMR) silages prepared with whole-plant corn in Tibet. TMR (354 g/kg DM) was ensiled with four different treatments: no additive (control), molasses (M), propionic acid (P), and molasses+propionic acid (PM), in laboratory silos (250 mL) and fermented for 45 d. Silos were opened and silages were subjected to an aerobic stability test for 12 days, in which chemical and microbiological parameters of TMR silages were measured to determined the aerobic deterioration. After 45 d of ensiling, the four TMR silages were of good quality with low pH value and ammonia/total N (AN), and high lactic acid (LA) content and V-scores. M silage showed the highest (p<0.05) LA content and higher dry matter (DM) recovery than the control and P silages. P silage had lower (p<0.05) LA content than the control silage. During aerobic exposure, lactic acid contents decreased gradually in the control and M silages, while that of P and PM silages increased, and the peak values were observed after 9 d. M silage had similar yeast counts with the control silage (> $10^5$ cfu/g FM), however, it appeared to be more stable as indicated by a delayed pH value increase. P and PM silages showed fewer yeasts (< $10^5$ cfu/g FM) (p<0.05) and were more stable than the control and M silages during aerobic exposure. It was concluded that M application increased LA content and improved aerobic stability of TMR silage prepared with whole-plant corn in Tibet. P application inhibited lactic acid production during ensiling, and apparently preserved available sugars which stimulated large increases in lactic acid during aerobic exposure stage, which resulted in greater aerobic stability of TMR silage.

• Journal of Microbiology and Biotechnology
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• v.20 no.11
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• pp.1563-1570
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• 2010

To construct a highly sensitive detection system for endocrine disruptors (EDs), we have compared the activity of promoters with the n-alkane-inducible cytochrome P450 gene (ALK1), isocitrate lyase gene (ICL1), ribosomal protein S7 gene (RPS7), and the translation elongation factor-1${\alpha}$ gene (TEF1) for the heterologous gene in Yarrowia lipolytica. The promoters were introduced into the upstream of the lacZ or hERa reporter genes, respectively, and the activity was evaluated by ${\beta}$-galactosidase assay for lacZ and Western blot analysis for hER${\alpha}$. The expression analysis revealed that the ALK1 and ICL1 promoters were induced by n-decane and by EtOH, respectively. The constitutive promoter of RPS7 and TEF1 showed mostly a high level of expression in the presence of glucose and glycerol, respectively. In particular, the TEF1 promoter showed the highest ${\beta}$-galactosidase activity and a significant signal by Western blotting with the anti-estrogen receptor, compared with the other promoters. Moreover, the detection system was constructed with promoters linked to the upstream of the expression vector for the hER${\alpha}$ gene transformed into the Y. lipolytica with a chromosome-integrated lacZ reporter gene under the control of estrogen response elements (EREs). It was indicated that a combination of pTEF1p-hER${\alpha}$ and CXAU1-2XERE was the most effective system for the $E_2$-dependent induction of the ${\beta}$-galactosidase activity. This system showed the highest ${\beta}$-galactosidase activity at $10^{-6}\;M\;E_2$, and the activity could be detected at even the concentration of $10^{-10}\;M\;E_2$. As a result, we have constructed a strongly sensitive detection system with Y. lipolitica to evaluate recognized/suspected ED chemicals, such as natural/synthetic hormones, pesticides, and commercial chemicals. The results demonstrate the utility, sensitivity, and reproducibility of the system for identifying and characterizing environmental estrogens.

• The Korean Journal of Nuclear Medicine
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• v.27 no.1
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• pp.123-129
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• 1993

Chitosan is known to be one of the natural chelating agents. It is derived from chitin which is a cellulose-like biopolymer distributed widely in nature, especially in shellfish, insects, fungi, and yeast. There are two forms of chitosans, water soluble and insoluble, The purpose of the present study is to investigate whether water soluble chitosan can be applied to reduce the bioabailability of radios-trontium in foods. We compared the effect of water soluble and insoluble chitosans on the absorption of ingested radiostrontium ($^{85}Sr$). Three percent water soluble and insoluble chitosan solutions were given orally, and immediately after $^{85}SrCl_2$ ($0.2{\mu}Ci$) was administered to rats using a orogastric tube. In one group water soluble chitosan solution was given for additional 4 days. And in control group no chitosan was given. Each group consisted of 6 rats. The whole-body retention of $^{85}Sr$, determined by in vivo counting method, was lower in water soluble chitosan group than that of water insoluble chitosan group and that of control. Urinary excretion of $^{85}Sr$ in chitosan-treated rats was higher than that of control. And 5 day ingested group of water soluble chitosan showed least whole body retention of $^{85}Sr$. In conclusion water soluble chitosan was more effective in reducing bioavailability of ingested radiostrontium in the gastrointestinal tract than insoluble chitosan.

• Advances in Traditional Medicine
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• v.5 no.3
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• pp.216-230
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• 2005

Although the field of study in immune enhancing compounds is relatively new, natural products from plants represent a rich and promising source of novel molecules with immunomodulating properties, Microglial cells, the main immune effector cells of the brain, usually display a ramified morphology and low expression levels of immunologically relevant antigens such as MHC class I and class II. Since any compound which participates in activation of phagocytic cells contributes to the production of potentially toxic factors, the search for convenient in vitro test-systems and study of mechanisms of action of these agents are of great interest. Human blood polymorphonuclear (PMN) cells and primary microglial cells isolated from Sprague-Dawley rats were used as cellular screening tests for study of phagocytosis-stimulating action of immunomodulating agents. Numbers of phagocytic activity were evaluated by the phagocyte ingestion of yeast cells and NO-synthase activity, nitrite production, and nitroblue tetrazolium test were determined after phagocyte stimulation. It was possible to demonstrate that indexes of phagocytic activity can be used as quantitative indicators for measurement immunomodulating activity. As a positive control, Zymosan A-induced phagocytosis in both PMN cells and primary microglial cells was used. $IFN-{\gamma}$ (0.1 -1 U/ml) stimulated phagocytosis in PMN cells 1.2 times after 2 - 3 h incubation, although at higher concentrations (10 - 100 U/ml) it strongly inhibited phagocytosis. In a similar way, at higher concentrations, $IFN-{\gamma}$ (100 - 500 U/ml) suppressed phagocytosis in zymosan-A stimulated microglial cells. When Polypodium leucotomus, cambricum and vulgare extracts were tested alone, increased levels of phagocytosis were observed in PMN. In addition, microglial cells showed both increased phagocytosis and MHC class-II antigen expressions. Surprisingly, when PMN and microglia were treated with a combination of Polypodium and $IFN-{\gamma}$, phagocytosis was not inhibited. We did not find changes in NO-synthase activity and nitrite production in both microglia and PMN cells activated by different immunomodulating agents. These results indicate that primary microglial cell cultures as well as human PMN cells can provide reproducible quantitative results in screening phagocytic activity of different immunoactive compounds. Furthermore, both inhibitory or activation mechanisms might be studied using these in vitro experimental approaches.

• Food Science and Preservation
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• v.11 no.4
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• pp.530-537
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• 2004

This study was conducted to determine the inactivation effect of electrolyzed water and organic acids either alone or in combination on L. monocytogenes or natural microflora on lettuce. Acidic electrolyzed water completely inactivated L. monocytogenes in broth system within 60 sec, but alkalin electrolyzed water caused approximate 1.7 log CFU/g reduction. However, acidic electrolyzed water reduced only 2.5 log CFU/g of L. monocytogenes on lettuce, and similar antimicrobial effect was observed with alkalin electrolyzed water. In the meantime, acidic and alkaline electrolyzed water caused approximately 2 log CFU/g reduction compared to control, whereas both electrolyzed water combined with $1\%$ organic acids ranged from 2.6 to 3.7 log CFU/g reduction. Among the organic acids, both electrolyzed water combined with $1\%$ citric acid showed the strongest synergistic antimicrobial effect to reduce L. monocytogenes on lettuce as well as total counts, yeast and molds. When antimicrobials, alone or in combination were treated into L. monocytogenes inoculated lettuce at $5^{\circ}C\;and\;15^{\circ}C$ for designed periods, the combined alkalin electrolyzed water with $1\%$ citric acid showed the greatest potential to inhibit growth of the bacteria. According to Scanning Electron Microscopy(SEM), the treatment of electrolyzed alkali water in combination with $1\%$ citric acid highly reduced the growth of the L. monocytogenes compared to single treatment and resulted in causing the destruction of cell membrane.

• Journal of the Korean Society of Food Science and Nutrition
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• v.30 no.4
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• pp.692-696
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• 2001

To develop natural food preservatives, ethanol and water extracts were prepared from the coriander (Coriandrum sativum L.) and antimicrobial activities were examined against 10 microorganisms which were food borne pathogens and/or food poisoning microorganisms and food-related bacteria and yeasts. Ethanol extract exhibited antimicrobial activities for the microorganisms tested, but not on lactic acid bacteria and yeast. Especially, minimum inhibitory concentrations (MIC) for Bacillus subtilis and Bacillus cereus were as low as 0.25mg/mL. antimicrobial activity of the ethanol extract was not destroyed by the heating at 121$^{\circ}C$ for 15min and not affected by pH. The ethanol extract of coriander exhibiting high antimicrobial activities was fractionated in the order of hexane, chloroform, ethylacetate and butanol fractions to test antimicrobial activity. The highest antimicrobial activity adjust bacteria tested was found in the ethylacetate fraction.

• Journal of the Korean Society of Food Science and Nutrition
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• v.21 no.1
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• pp.97-100
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• 1992

The optimum condition for the extraction of antimicrobial substance from Lithospermi radix was investigated. For the purpose of obtaining basic data for the development of natural preservatives and for the prevention of food poisoning accidents, its antimicrobial activity was tested against several kinds of saprophytic microbes and food poisoning bacteria. The optimum condition for extraction of antimicrobial substance was to steep Lithospermi radix into 95% ethanol for 24 hours at room temperature. Antimicrobial activity was observed at the pH range $5.0{\sim}8.0$, but its activity became stronger at acidic condition. The result of ion exchange chromatography was showed that the antimicrobial activity of anionic portion was more apparent than that of cationic portion. The antimicrobial activity against Gram positive bacteria was stronger than that of Gram negative bacteria and the growth of food poisoning bacteria such as S. aureus and V. parahaemolyticus was inhibited in the concentration of 0.1% for 48 hours. As for mold and yeast, the growth of some kinds of these organisms was inhibited in the concentration of 0.1 % for 48 hours and the growth of nearly all the fungi was inhibited in the concentration of 0.15% for 96 hours.

• Journal of Life Science
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• v.20 no.5
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• pp.683-690
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• 2010

The yeast strains of Saccharomyces diastaticus produce one of three isozymes of an extracellular glucoamylase I, II or III, a type of exo-enzyme which can hydrolyse starch to generate glucose molecules from non-reducing ends. These enzymes are encoded by the STA1, STA2 and STA3 genes. Another gene, sporulation-specific glucoamylase (SGA), also exists in the genus Saccharomyces which is very homologous to the STA genes. The SGA has been known to be produced in the cytosol during sporulation. However, we hypothesized that the SGA is capable of being secreted to the extracellular region because of about 20 hydrophobic amino acid residues at the N-terminus which can function as a signal peptide. We expressed the cloned SGA gene in S. diastaticus YIY345. In order to compare the biochemical properties of the extracellular glucoamylase and the SGA, the SGA was purified from the culture supernatant through ammonium sulfate precipitation, DEAE-Sephadex A-50, CM-Sephadex C-50 and Sephadex G-200 chromatography. The molecular weight of the intact SGA was estimated to be about 130 kDa by gel filtration chromatography with high performance liquid chromatography (HPLC) column. Sodium dedecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed it was composed of two heterogeneous subunits, 63 kDa and 68 kDa. The deglycosylation of the SGA generated a new 59 kDa band on the SDS-PAGE analysis, indicating that two subunits are glycosylated but the extent of glycosylation is different between them. The optimum pH and temperature of the SGA were 5.5 and $45^{\circ}C$, respectively, whereas those for the extracellular glucoamylase were 5.0 and $50^{\circ}C$. The SGA were more sensitive to heat and SDS than the extracellular glucoamylase.

• The Korean Journal of Mycology
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• v.19 no.1
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• pp.54-60
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• 1991

In the traditional medicine, the basidiocarps of Poria cocos (Fr.) Wolf naturally collec­ted have been widely used as a traditional remedy for dropsy, diarrhea, gonorrhea and nervous disease more than 3500 years ago. It was recently found to be composed of the particular compounds having antitumor effects. Therefore, its demand has been gradually increased, whereas it is still dependent upon the natural harvest. The artificial cultivation of P. cocos was carried out to observe the morphological characteristics and to investigate the cultural characteristics of P. cocos isolates collected. The morphological characteristics of its basidiocarps was observed. The physiological aspects of its isolates were also investigated.