• The Journal of the Korean Society for Microbiology
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• v.21 no.1
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• pp.151-161
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• 1986

In order to develop sensitive and sepcific assay methods for E. coli heat labile enterotoxin(LT) hybridoma cell lines secreting LT specific monoclonal antibody were obtained. LT was purified from cell lysate of E. coli O15H11. The steps included disruption of bacteria by French pressure, DEAE Sephacel ion exchange chromatography, Sephadex G200 gel filtration, and second DEAE Sephacel ion exchange chromatography, successively. Spleen cells from Balb/c mice immunized with the purified LT and $HGPRT^{(-)}$ plasmacytomas, $P3{\times}63Ag8.V653$ were mixed and fused by 50% (w/v) PEG. Hybrid cells were grown in 308 wells out of 360 wells, and 13 wells out of them secreted antibodies reacting to LT. Among these hybridoma cell 1G8-1D1 cell line was selected since it had produced high-titered monoclonal antibody continuously. By using culture supernatant and ascites from 1G8-1D1 cells the monoclonal antibody was characterized, and an assay system for detecting enterotoxigenic E. coli was established by double sandwich enzyme-linked immunosorbent assay (ELISA). The following results were obtained. 1. Antibody titers of culture supernatant and ascites from 1G8-1D1 hybridoma cells were 512, and 102, 400, respectively by GM1-ELISA and its immunoglobulin class was IgM. 2. The maximum absorption ratio of 1G8-1D1 cell culture supernatant to LT was 90% at $300\;{\mu}g/ml$ of LT concentration. LT concentration shown at 50% absorption ratio was $103.45{\mu}g$ and the absorption ratio was decreased with tile reduction of LT concentration. This result suggests that monoclonal antibody from 1G8-1D1 hybridoma cell bound with LT specifically. 3. The reactivities of 1G8-1D1 cell culture supernatant to LT and V. cholerae enterotoxin(CT) were 0.886 and 0.142(O.D. at 492nm) measured by the GM1-ELISA, indicating 1G8-1D1 monoclonal antibody reacted specifically with LT but not with CT. 4. The addition of 0.1ml of ascites to 0.6mg and 0.12mg of LT decreased the vascular permeability factor to 41% and 44% respectively, but it did not completely neutralize LT. 5. By double sandwich ELISA using monoclonal antibody, as little as 75ng of the purified LT per ml could be detected. 6. The results by assay of detecting LT in culture supernatants of 14 wild strains E. coli isolated from diarrhea patients by the double sandwich ELISA were almost the same level as those by reverse passive latex agglutination.

• Journal of Bio-Environment Control
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• v.11 no.4
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• pp.181-187
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• 2002

The overall objective of this study was to improve tomato fruit quality, while maximizing yield. The variety of 'Momotaro' was grown in the basic nutrient solution of 1.6 dS.m$^{[-10]}$ which was supplemented by three levels of seawater with EC 1.0, 2.0 or 3.0 dS.m$^{[-10]}$ . Tomato plants were cultivated in cool seasons. Plant growth characteristics were compared between treatments, and fruits were classified to analyse fruit quality characteristics according to ripening stages： MG, Br, Br＋3, Br＋5, Br＋7 and Br＋10. Adding seawater generally did not affect the shoot growth parameters such as plant height, leaf length, leaf width, internode length and chlorophyll content. Adding seawater negatively affected yield parameters such as the height and weight of fruit, marketable fruit weight per plant and marketable fruit yield. Therefore, the more yield reduction was obtained with the increasing level of seawater treatment. Fruit quality was improved by seawater treatment. The degree of the effect for $^{\circ}$Bx degree and sugars were the highest with the EC of seawater 2.0~3.0 dS.m$^{[-10]}$ , and at the Br＋5~Br＋7 of ripening stages. The relative abundance of tomato flavor, volatile components, was not generally affected by the seawater treatment with an exception of 6-methyl-5-hepten-2-one. The relative abundance of most volatile components increased as ripening progressed. The increment began at the Br stage and showed the highest increment at the Br＋5~Br＋7 stages. The results from these experiments suggest that seawater treatment of EC 3.6 dS.m$^{[-10]}$ for hydroponics is good for improving tomato quality. Fruit quality is the best at the Br＋5~Br＋7 ripening stages. It is considered that these results may be applied far use in hydroponic culture to improve fruit quality with minimum yield reduction.

• Tuberculosis and Respiratory Diseases
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• v.43 no.5
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• pp.728-735
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• 1996

Background : Recognition and ingestion of opsonized microorganisms by neutrophils induces the burst of oxidative metabolic activity. Products of the respiratory burst activity provide powerful oxygen dependent killing mechanism. Measurement of respiratory burst activity has been a major indicator of the functional capacity of neutrophils. We determined the respiratory burst activity of neutrophils in patients with pneumonia and observed the changes during the clinical course of pneumonia. Methods: The EDTA blood was drawn from 24 normal controls and same numbers of pneumonia patients. The respiratory burst activity(with the production of $H_2O_2$ which changes nonfluorescent DCF-DA to green fluorescent DCF) in the non-stimulated state and the stimulated state with fMLP and PMA of neutrophils was measured by flowcytometry at day 1, 3, 5, 7 and 9 of admission. Results: The respiratory burst activity of neutrophils was mildly increased by stimulation with fMLP. But there was no statistical significance between normal control and patients with pneumonia. The respiratory burst activity of neutrophils was markedly increased by stimulation with PMA in both groups. There was a significant difference in response to PMA between normal control and patients with pneumonia. The production of hydrogen peroxide from neutrophils was decreased during early course of pneumonia and it was recuperated gradually to normal level in 9 days. Conclusion : Hydrogen peroxide production from neutrophils was suppressed during early course of pneumonia and restored after treatment. It is suggested that the production of oxygen radical in response to PMA stimulation from each neutrophils is decreased rather than increased during the early course of pneumonia.

• Food Science and Preservation
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• v.23 no.6
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• pp.797-803
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• 2016

In this study, we investigated the effect of fermentation conditions on the amylolytic and proteolytic activities of Aspergillus luchuensis strain 74-5 and Aspergillus oryzae strain 75-2, which are used in the preparation of the starter culture, for Takju (Korean traditional rice wine). The starter culture was optimized using different conditions, such as inoculum size, inoculation temperature, and incubation time. The enzyme activities under each condition were measured. In the A. luchuensis strain 74-5 starter culture, the ${\alpha}-amylase$ and glucoamylase activities increased, however the activity of acidic protease decreased as the diluent to starter culture ratio increased. In the A. oryzae 75-2 starter culture, all enzyme activities were maintained at a higher level even at 5% inoculation ratio. Higher enzyme activities were observed in the middle range of inoculation temperature (35, $40^{\circ}C$), than in the lower range (20, $30^{\circ}C$). Enzyme activity in the starter culture varied with incubation time, however it was the highest at 144 and 120 hr, respectively, for A. luchuensis strain 74-5 and A. oryzae strain 75-2. The spore count of the starter culture was approximately $2{\times}10^7$ during fermentation, out of which contamination by aerobic bacteria was about $3{\times}10^3$. The results suggested that the starter culture of each strain could be used as an inoculum for fermentation. However, we needs to conduct further research for the selection of suitable diluting agents as well as drying methods to reduce the contamination by aerobic bacteria, while retaining the enzyme activity.

• Journal of Korean Society of Forest Science
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• v.35 no.1
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• pp.1-8
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• 1977

Forest soil fertility was evaluated through the approach of numerical method. In this study, the soil chemical properties analyzed for 35 different soil series as table 2 were cited in numerical analysis. Minimum contents of essential nutrients in the surface soil for a satisfactory growth of tree in the plantation were evaluated by comparing with Wild's standard as table 1. Demanding level of fertilization were evaluated by using the formula 1 as table 5. Similar relation of soil chemical properties between soil series were calculated through formula 2, and then classified into 5 groups in soil chemical properties. 1. General chemical properties of surface soil in case of 35 soil series. About 40 percent of 35 different soil series are less than 2 percent in organic matter, 10 ppm in available phosphorus, 1.25m.e/l00g in exchangeable calcium and 0.5m.e/l00g in exchangeable magnesium. Generally, shortage of exchangeable potash are not found. CEC less than 10m.e/l00g are in two thirds and strong acid soil less than PH 5.5 are in about four fifths. 2. Soil series requested or not the fertilization are indirectly evaluated from the formula 1 using the relative figure of chemical components of CEC, OM and MgO. Through this analysis, 8 different soil series have very poor quality in soil chemical capacity so that demands highly the fertilization. On the other hand, other 13 different soil series group have not been thought to need the fertilization according to chemical guality. 3. By the results comparing the similarity of chemical properties of forest soil, it is thought to be suitable that the forest soil fertility are divided into 5 groups as follows: 1. Low CEC soil 1-1 Low organic matter soil less than 2 percent 1-2 Medium organic matter soil less than 4 percent 2. High CEC and organic matter soil 2-1 Low magnesium soil 2-2 High magnesium soil 3. High magnesium and calcium soil as lime stone.

• Journal of The Korean Society of Grassland and Forage Science
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• v.31 no.4
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• pp.395-408
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• 2011

The study which it sees exclusions the copper and the zinc which contain in pig sludge, It study pig sludge resources fertilizer production which are rational, pig sludge resources fertilizers after seeding, silage corn it investigates growth characteristics and forage value, the result which investigates pig sludge resources fertilizers effectiveness with afterwards is same. With fertilizer ingredients in pig sludge chemical qualities, the content of the nitrogen and the phosphoric acid comes 4.4% to be 6.29%, pH 7.02 and content of the copper and the zinc which is a heavy metal which contains in pig sludge with 805 mg/kg and 1,704 mg/kg, it is a restrictive standard of the fertilizer, 300 mg/kg and 900 mg/kg it sees to be high, it manufactures citric acid 1 hydrate with the organic acid solution, heavy metals of pig sludge where it is a mixture ratio of the organic acid solution, it divides to 25%, 50%, 75% and 100% 4 kind levels, the result which measures the heavy metal exclusion ratio of the copper and the zinc, the mixture ratio of the organic acid solution to be many exclusion ratio of the copper and the zinc is showing a just interrelation, from organic acid solution 100% level content of pig sludge remains copper and zinc 330.03 mg/kg and 41.28 mg/kg, it shows the exclusion ratio of copper 59% zinc 97%. 'Cheonganok' growth characteristics with citric acid 1 hydrate, Treatment 2 and control growth characteristics etc, it exclusion the copper and the zinc it doesn't appear on significant difference statistically but, treatment 3 after only pig sludge in resources disposal where it seeding, growth characteristics of leaf area etc. is badness, it compares in control and treatment 2 the growth characteristics badness, it is appearing, it is caused by with disease and insects occurrence of $Ostrinia$ $furnacalis$ and brown spot, the damage was many. From forage value, Treatment 2 where it exclusion the heavy metal with the citric acid 1 hydrate with control it compares and there are not significant difference from crude protein and ADF and NDF contents etc., seeding only Pig Sludge in resources disposal treatment 3, it is caused by with $Ostrinia$ $furnacalis$ etc., trunk and aging of the leaf to be high ADF content is low. but from crude protein, the nitrogen ingredient which pig sludge has and interrelation it seemed and high numerical value were confirmed.

• Journal of Life Science
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• v.17 no.10
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• pp.1309-1314
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• 2007

Investigation on health conditions of Thai koi (Anabas testudineus) were carried out through clinical and histopathological observations from different farms of Mymensingh district for seven months during August 2006 to February 2007. Fish sampling and water quality parameters (temperature, dissolved oxygen and pH) were monitored on a monthly basis. Clinical examination of fishes was also carried out for any kind of abnormalities at monthly intervals. Samples of skin, muscle, gill, liver and kidney were observed by histological technique. Among the water quality parameters the values of water temperature, dissolved oxygen were found to be at unfavorable level for fish during the colder months in the farms. Clinically it was observed that fishes were more affected from December to January and almost normal in appearance during August to September and February. Different clinical symptoms like scale loss, dermal lesion, ulcer and loss of caudal fin were noticed in December and January. In histopathological study, structures of fish organs were normal from August to September. In the months of October and November, minor pathologies were found to be started. Marked pathological changes like necrosis, pyknosis, inflammation, hemorrhage, hypertrophy, hyperplasia, missing of primary and secondary gill lamellae were observed in the months of December and January. Whereas, in the month of February the pathological condition of fish gradually reduced. Again when considered individual fish farm, fishes of Reliance Aqua Farm were more affected than Sotota Matshya Hatchery. The study showed that severity of clinical and pathological changes were increased in December and January. During the period EUS and protozoan diseases were noticed in A. testudineus of the investigated farms.

• KSBB Journal
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• v.21 no.3
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• pp.204-211
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• 2006

For the production of the recombinant human interferon-gamma(rhIFN-${\gamma}$) in Escherichia coli, human glucagon and ferritin heavy chain were used as fusion partners. Even though rhIFN-${\gamma}$ is expressed as an inclusion body form in E. coli because of strong hydrophobicity of itself, over 50% of fused rhIFN-${\gamma}$ was expressed as soluble form in E. coli $Origami^{TM}$(DE3) harboring pT7FH(HE)-IFN-${\gamma}$ which encodes ferritin heavy chain-fused rhIFN-${\gamma}$. In the case of using glucagon-ferritin heavy chain hybrid mutant as a fusion partner, 6X His-tag was additionally introduced to N-terminus of GFHM(HE)-IFN-${\gamma}$ for enhancing purification yields of rhIFN-${\gamma}$. Fusion protein HGFHM(HE)-IFN-${\gamma}$ with two 6X His-tag was more effectively bound to Ni-NTA agarose bead than GFHM(HE)-IFN-${\gamma}$ with a 6X His-tag. rhIFN-${\gamma}$ was completely purified from enterokinase-treated HGFHM(HE)-IFN-${\gamma}$ by Ni-NTA affinity column. For high-level production of rhIFN-${\gamma}$, glucose was used as the sole carbon source with simple exponential feeding rate($2.4{\sim}7.2g/h$) in fed-batch process. The effective lactose concentration for the expression of the rhIFN-${\gamma}$ was $10{\sim}20mM$. Under the fed-batch culture conditions, rhIFN-${\gamma}$ production yield reached 11 g DCW/L for 6 hours after lactose induction.

• The Korean Journal of Pesticide Science
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• v.19 no.1
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• pp.5-13
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• 2015

A chromatographic method for the determination of imazapyr, a non-selective herbicide, in agricultural commodities was developed to use safety control of pesticide residue on crops, and was fully validated as an official method for residue analysis. Agricultural commodities, mandarin (fruit), hulled rice (cereal grains), pepper (vegetables), potato (potatoes) and soybean (beans) were extracted with methanol and partitioned with dichloromethane to remove the interference obtained from sample extracts, adjusting pH to 2.5 by 4N hydrochloric acid. Finally, they were analyzed by high performance liquid chromatography coupled to UV detector (HPLC-UVD). The developed method had the linearity in the range of test concentrations with coefficients of determination ($r^2$) more than 0.99. Recovery studies were carried out at three concentration levels (LOQ, 10LOQ, and 50LOQ) performing five replicates at each level. Recoveries were ranged between 72.1 to 108.0%, with relative standard deviations less than 10%. A consistent recovery was determined according to the CODEX guidelines (CAC/GL40, 2003). Finally, LC/MS with selected ion monitoring was also applied to confirm the suspected residues of imazapyr in agricultural samples. This developed method for determination of imazapyr residues in agricultural commodities. can be used as an official method.

• Journal of Life Science
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• v.29 no.8
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• pp.904-915
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• 2019

Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.