Recently, the demand for the cultivation of upland soil has been increasing, and the rate of conversion of paddy soil into upland soil is also increasing. Theincrease in uneven precipitation due to climate change has resulted in dramatic effects of waterlogging stress on upland crops. Therefore, the present study was conducted to investigate the changes in growth characteristics and the expression patterns of proteins at the two-leaf stage of adzuki beans. The domestic cultivar, Arari (Miryang No. 8), was used to test waterlogging stress. At the two-leaf stage of adzuki beans, plant height slightly decreased androot fresh weight showed significant changes after 3 days of waterlogging treatment. Chlorophyll content was also significantly different after 3 days of waterlogging treatment compared to its content in control plants. Using two-dimensional gel electrophoresis, more than 400 protein spots were identified. Twenty-one differentially expressed proteins from the two-leaf stage were analyzed using linear trap quadrupole-Fourier transform-ion cyclotron resonance mass spectrometry. Of these 21 proteins, 9 were up-regulated and 12 were down-regulated under waterlogging treatment. Protein information resource (https://pir.georgetown.edu/) categories were assigned to all 49 proteins according to their molecular function, cellular component localization, and biological processes. Most of the proteins were found to be involved in the biological process, carbohydrate metabolism and were localized in chloroplasts.
Bacteriostatic activity of secretory immumoglobulin A (SIgA) in human and bovine colostrums on enterotoxigenic type Salmonella was tested in the tissue culture medium. SIgA was used in $0.1{\sim}5.0mg/m{\ell}$ concentration with or without the addition of egg lysozyme tested for theirs bacteriostatic activites. 1. Bovine SIgA rich fraction with a large amount of $IgG_1$-dimer could be prepared from bovine colostrum of Holstein cows by anion exchange chromatography using DEAE-Sephadex A-50 and gel filtration on Sephadex G-200 and Sepharose 6B. 2. Human SIgA appeared to be the most bacteriostatic effect for all varieties of Salmonella in a range of $0.5{\sim}1.0mg/m{\ell}$ Bovine SIgA showed a marked bacteriostatic effect increased by increasing the concentration. Bovine IgG had not show bacteriostatic effect against both enterotoxigenic type Salmonella. Egg lysozyme as well as bovine SIgA also showed a marked bacteriostatic effect increased by increasing the concentration. 3. When the growth inhibition of human SIgA was tested by adding egg lysozyme with time interval, egg lysozyme showed bacteriostatic effect as compared with control. But human SIgA and adding with lysozyme showed slight the bacteriostatic effect. 4. When the growth inhibition of bovine SIgA was tested by adding egg lysozyme with time interval, all treatment against S. enteritidis showed bacteriostatic effect as compared with control In the case of S. typhinwrium, egg lysozyme showed a marked bacteriostatic effect as compared with control.
Kim, Kwang-Joo;Kim, Hee-Kyung;Park, Young-Ran;Park, Jae-Yun
Journal of the Korean Magnetics Society
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제16권1호
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pp.23-27
/
2006
By substituting Mn or Cr for Co in inverse spinel $CoFe_2O_4,\;Mn_xCo_{1-x}Fe_2O_4\;and\;Cr_xCo_{1-x}Fe_2O_4$ and thin films were prepared by sol-gel method and their structural and magnetic properties were investigated. X-ray diffraction indicates that the cubic lattice constant increase for the Mn substitution while it hardly changes for the Cr substitution. Substitution of $Mn^{2+}$ for octahedral $Co^{2+}$ sites can explain the increase of lattice constant in $Mn_xCo_{1-x}Fe_2O_4$. On the other hand, Substitution of $Cr^{3+}$ for octahedral $Co^{2+}$ and subsequent reduction of $Fe^{3+}$ ion into $Fe^{2+}$ are expected to happen. Mossbauer spectroscopy measurements on $Cr_xCo_{1-x}Fe_2P_4$ indicate the existence of tetrahedral $Fe^{2+}$ ions that are created through reduction of tetrahedral $Fe^{3+}$ ions in order to compensate charge imbalance happened by $Cr^{3+}$ substitution for octahedral $Co^{2+}$ sites. On the other hand, no $Fe^{2+}$ ions were detected by Mossbauer spectroscopy for $Mn_xCo_{1-x}Fe_2O_4$. A migration of $Fe^{3+}$ ions from octahedral to tetrahedral sites In $Mn_xCo_{1-x}Fe_2O_4$ was detected by Mossbauer spectroscopy for x>0.47. Vibrating sample magnetometry measurements on the samples at room temperature revealed that the saturation magnetization increases by Mn and Cr substitution for certain range of x, qualitatively explainable in terms of the comparison of spin magnetic moment among the related transition-metal ions.
The characteristics of the three alkaline proteinases, Enz. A, B and C, from the pyloric caeca of mackerel have been investigated. The optimum condition for the activity of the Enz. A, B and C was pH 9.4, 9.8 and 9.8 at $45^{\circ}C$ for $2\%$ casein solution, and was pH 9.2 10.2 and 9.8 at $45^{\circ}C$ for $5\%$ hemoglobin denatured by urea, respectively. Enz. A, B and C by heat treatment at $50^{\circ}C$ for 5 min were inactivated 90, 33 and $37\%$, respectively, over the original activity. The reaction rate of the three alkaline proteinases was constant to the reaction time to 40 min in the reaction condition of $2{\mu}g/ml$ of enzyme concentration and $2\%$ casein solution. The reaction rate equation and Km value against casein substrate determined by the method of Lineweaver and Burk were: Enz. A, Y=3.6X and $Km=5.0{\times}10^{-3}\%$; Enz. B, Y=6.0X and $Km=1.0{\times}10^{-3}\%$; Enz. C, Y=4.2X and $Km=3.6{\times}10^{-3}\%$. The three alkaline proteinases were inactivated by $Ag^+$ and $Hg^{2+}$, but activated by $Mn^{2+},\;Sn^{2+}\;and\;Pb^{2+}$, Enz. B and C were remarkably inhibited by the soybean trypsin inhibitor. Molecular weight of Enz. A, B and C determined by SDS-polyacrylamide gel electrophoresis and Sephadex G-100 gel filtration was in the range of $27,500{\pm}2,500,\;20,500{\pm}1,500\;and\;15,250{\pm}250$, respectively.
Hyun Bek;Lim Hak-Seob;Chung Kyung Kae;Choi Yung Hyun;Choi Byung Tae;Seo Min-Jeong;Kim Ji-Eun;Ryu Eun-Ju;Huh Man Kyu;Joo Woo Hong;Jeong Young Kee
Journal of Life Science
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제15권6호
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pp.987-993
/
2005
A bacterium, producing a fibrinolytic enzyme, was screened from a decaying rice plant. The bacterium was identified as Bacillus subtilis by morphological, biochemical, and physiological properties and named Bacillus subtilis BK-17. The fibrinolytic enzyme (BK) was purified from supernatant of Bacillus subtilis BK-17 culture broth. The molecular weight was 31 kDa as determined by SDS-PAGE. The effect of temperature, pH, and plasminogen on the activity of the bacillokinase (BK) was analysed and the activity was compared with urokinase. The optimal temperature and pH were $50^{circ}C$ and pH 7, pH 8, respectively. The BK activity was inhibited to $45\%$, $35\%$, and $23\%$ with 1mM EDTA, $Zn^{2+}$, and $Ca^{2+}$, respectively. However, $Mg^{2+}$, $Mn^{2+}$, and $Co^{2+}$ ions did not have any significant effect on the enzyme activity The BK showed the artivity in the both plates, plasminogen-free fibrin plate and plasminogen-rich fibrin plate. The result indicates that the BK can directly act the fibrin. In comparison of fibrinolytic activity with urokinase on the fibrin plate, the BK shows about 20 folds higher activity than that of the urokinase.
In tile Iron-catalyzed Haber-Weiss reaction to produce OH., the requirement for $O^{-}_{2}{\cdot}$ is only to reduce $Fe^{+++}$. Possibly, the role of $O^{-}_{2}{\cdot}$ can be replaced by other reducing agents. Ascorbate is one of them in biological system. In the present study, the ability of ascorbate to produce $OH{\cdot}$ in the presence of $Fe^{++}$ and $H_2O_2$ was investigated by observing the degradation of hyaluronic acid and ethylene production from methional. Ascorbate stimulated the degradation of hyaluronic by $Fe^{++}$ and $H_2O_2$. That was confirmed by both viscosity change and gel-permeation chromatographic analysis. The observed degradation was almost completely prevented by catalase and $OH{\cdot}$ scavengers. In support of the above results, ascorbate enhanced the prouction of ethylene from methional in the presence of $Fe^{++}$ and $H_2O_2$. Other reducing agents (cysteine, glutathione, NADH and NADPH) showed similar activities to ascorbate in the degradation of hyaluronic acid and ethylene production. But no stimulatory effects were observed with their oxidized forms such as NAD and NADP. Thus, it appears that reduction of the metal ion was needed for $OH{\cdot}$ production. Among the metal ions tested, $Fe^{++}$ showed most potent catalytic action in the production of $OH{\cdot}$ The results obtained support that ascorbate can substitute $O^{-}_{2}{\cdot}$ in the metal-catalyzed reactions, particularly with $Fe^{++}$ by which $OH{\cdot}$ is produced with $H_2O_2$. The significance of the ascorbate-dependent production of $OH{\cdot}$ was considered with respect to possible role of ascorbate in the damage of inflamed joints.
Ham, S.S.;Park, B.K.;Lee, S.Y.;Lee, J.H.;Kang, C.K.;Lee, D.S.;Omura, Hirohisa
Applied Biological Chemistry
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제27권4호
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pp.264-270
/
1984
In order to obtain mutagenic data of enzymatic browning reaction products, we investigated their mutagenicity. The rec-assay with Bacillus subtilis strains $H17(rec^+)$ and $M45(rec^-)$ were carried out with their spores. Detection of double strand breakage in calf-thymus DNA was investigated into sample solution with and without $Cu^{2+}$ by the agarose slab gel electrophoresis. In the rec-assay, catechol, pyrocatechol, DL-dopa, 3,4-dihydroxytoluene, hydroquinone, hydroxyhydroquinone with and without $Cu^{2+}$ in 0.05M ana 0.1M at the enzymatic browning reaction products stowed mutagenic action. And also browning solution of 0.05M hydroxyhydroquinone and catechol with $Cu^{2+}$, hydroquinone with and without $Cu^{2+}$ of 0.1M at the enzymatic browning reaction products were strong mutagenic action. The DNA breakability of the enzymatic browning reaction products of 0.1M pyrogallol was stronger than that of 0.05M pyrogallol browning solution with $Cu^{2+}$ and 3,4-dihydroxytoluene browning solution was stronger than that of 0.01M 3,4-dihydroxytoluene browning solution.
Kim, Cheol-Min;Park, Seung-Kyu;Shon, Mal-Hyun;Song, Sun-Dae;Kim, Young;Jun, Eun-Sook;Son, Han-Chul;Jung, Byung-Sun
Tuberculosis and Respiratory Diseases
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제43권1호
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pp.30-37
/
1996
Background: The extraction methods of DNA from clinical samples are the major obstacle to use the PCR(polymerase Chain Reaction) in routine labortary for early detection of M. tuberculosis. We tried to improve the extraction method of DNA from sputum for establishment of the PCR in routine labortary by reducing the possibility of cross contamination and performing it easily and safely. Methods: We used the $InstaGene^{TM}$ DNA extraction kit(BioRad Co.) using Chelex 100 ion exchange resin for preparation of DNA. We compared InstaGene method in 100 cases of sputum from proteinase K method which is known as the most commonly used method for DNA purification(Experiment 1). And we compared InstaGene method in 98 cases of sputum from Microwave method developed by a company in Korea(Experiment 2). In experiment 1,245bps of IS6110 were amplified and then 188bps were amplified by nested PCR. In experiment 2,536bps in primary PCR and 276bps in nested PCR were amplified and analysed by agarose gel electrophoresis and EtBr staining. Results: When we chose AFB smear, culture, or AFB smear and culture as a standard test, PCR had low specificity and positive predictive value in both experiments. The InstaGene method has higher value in sensitivity and negative predictive value significantly than proteinase K method. The InstaGene method and the Microwave methods were similar in sensitivity, specificity, positive predictive value and negative predictive value. Conclusion: Even though both methods had lower possibility of cross contamination, shorter time requirement, simplicity, and economic advantages than Proteinase K method, the InstaGene method was a little simpler than the Microwave method. Therefore, in terms of usefulness in clinical application, the Instagene method seems to be the most useful method in DNA extraction for detection of M. tuberculosis using PCR. The reliability of this method will be clarified by further studies with enough clinical samples.
ZrO2 phase transformations depending on the type and amount of dopants and the sintering temperatures were studied for the 2 components (CaO-, Y2O3-, MgO-ZrO2) and the 3 components(MgO-ZrO2-Al2O3)ZrO2 powder by X-ray diffraction and Raman spectroscopy. In the CaO- and Y2O3-ZrO2 systems, as the CaO and Y2O3 contents increased to 6~15mol% and 3~15mol% respectively, we were not able to identify between tetragonal and cubic in the X-ray diffraction patterns. On the other hand, all Raman modes shifted to lower wavenumbers, decreasing in intensity and the number of bands, markedly. These phenomena were caused by tetragonallongrightarrowcubic phase transformation and interpreted by the breakdown of the wave vector selection rule(k=0) and the structural disorder associated with the formation of oxygen sublattice which was caused by the substitution between Zr4+ ion and Ca2+ or Y3+ ion in ZrO2 matrix. The monoclinic to cubic phase transformation occurred in 10mol% MgO-ZrO2 system. As the Al2O3 content increased from 0 to 20mol% in the MgO-ZrO2-Al2O3 systems, cubic phase transformed to monoclinic phase, this is because the MgO didn't play a role in a stabilizer because of the formation of the spinel(MgAl2O4) by the reaction between MgO and Al2O3, Also, the ZrO2 phase transformation was explained by the change of it's lattice parameters depending on the type and amount of dopants. Namely, as the amount of dopant increased to 10~13mol%, the axial ra-tio c/a came close to unity with increasing the lattice parameter a and decreasing the lattice parameter c. At that time, the tetragonallongrightarrowcubic phase transformation occurred.
The Journal of the Korean Society for Microbiology
/
제21권1호
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pp.151-161
/
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.
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