• Journal of Microbiology and Biotechnology
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• v.31 no.5
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• pp.705-709
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• 2021

Porphyromonas gingivalis (P. gingivalis) is a major bacterial pathogen that causes periodontitis, a chronic inflammatory disease of tissues around the teeth. Periodontitis is known to be related to other diseases, such as oral cancer, Alzheimer's disease, and rheumatism. Thus, a precise and sensitive test to detect P. gingivalis is necessary for the early diagnosis of periodontitis. The objective of this study was to optimize a rapid visual detection system for P. gingivalis. First, we performed a visual membrane immunoassay using 3,3',5,5'-tetramethylbenzidine (TMB; blue) and coating and detection antibodies that could bind to the host laboratory strain, ATCC 33277. Antibodies against the P. gingivalis surface adhesion molecules RgpB (arginine proteinase) and Kgp (lysine proteinase) were determined to be the most specific coating and detection antibodies, respectively. Using these two selected antibodies, the streptavidin-horseradish peroxidase (HRP) reaction was performed using a nitrocellulose membrane and visualized with a detection range of 10³-10⁵ bacterial cells/ml following incubation for 15 min. These selected conditions were applied to test other oral bacteria, and the results showed that P. gingivalis could be detected without cross-reactivity to other bacteria, including Streptococcus mutans and Escherichia fergusonii. Furthermore, three clinical strains of P. gingivalis, KCOM 2880, KCOM 2803, and KCOM 3190, were also recognized using this optimized enzyme immunoassay (EIA) system. To conclude, we established optimized conditions for P. gingivalis detection with specificity, accuracy, and sensitivity. These results could be utilized to manufacture economical and rapid detection kits for P. gingivalis.

• Journal of the Korean Society of Food Science and Nutrition
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• v.32 no.4
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• pp.506-512
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• 2003

Peach wine was fermented at $25^{\circ}C$ for 2 weeks using Saccharomyces cerevisiae KCCM 12224, aged at 15$^{\circ}C$ for 14 weeks, and its physicochemical and microbiological changes were investigated. The viable bacterial cell numbers, 1.4$\times$10$^3$ CFU/mL at the beginning of fermentation, increased to 2.8$\times$10$^{6}$ CFU/mL after 2 weeks, but decreased to 7.0$\times$10$^3$ CFU/mL after 14 weeks. The viable yeast cell numbers were changed from 3.4$\times$10$^2$ CFU/mL to 2.4$\times$10$^{7}$ CFU/mL during fermentation, and decreased to 4.0$\times$10$^4$ CFU/mL after aging. Turbidity total sugar content, reducing sugar content, solid content and b value of peach wine decreased during fermentation but acidity, alcohol content, L and a value increased. Most physicochemical properties except alcohol content and reducing sugar content were not changed significantly during aging. When peach wine was filtered through 0.45 ${\mu}{\textrm}{m}$ nitrocellulose membrane followed by various ultrafiltration membranes with different molecular weight cut-off values, Biomax 100K membrane, with 79 liter/$m^2$/h (LMH) of initial flux, was suitable for ultrafiltration process of peach wine. These membrane filtration treatments resulted in complete removal of microorganisms and decrease in turbidity and alcohol content without changes in other chemical properties. The physicochemical properties of peach wine were not changed and any microorganisms were not found during the storage at 3$0^{\circ}C$ for 12 Weeks.

• Applied Biological Chemistry
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• v.46 no.3
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• pp.201-206
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• 2003

An apple wine was prepared by fermentation at $25^{\circ}C$ for 2 weeks using Saccha개myces cerevisiae KCCM 12224, followed by aging at $15^{\circ}C$ for 14 weeks, and its physicochemical and microbiological changes were investigated. The viable bacterial cell numbers, increased from $1.4{\times}10^3\;CFU/ml$ at the beginning of fermentation, to $2.8{\times}10^6\;CFU/ml$ after 2 weeks, but decreased to $1.0{\times}10^5\;CFU/ml$ after aging. The viable yeast cell numbers changed from $4.3{\times}10^4\;CFU/ml$ to $1.2{\times}10^7\;CFU/ml$ during the fermentation, and decreased to $1.2{\times}10^4\;CFU/ml$ after aging. Sugar content changed from $20.0^{\circ}Brix$ to $8.5{\circ}Brix$, and reducing sugar content was changed from 9.66% to 6.44%. Alcohol content and acidity increased to 7.0% and from 0.19% to 0.24%, respectively. No changes in acidity, pH, and sugar content were observed during the aging, but reducing sugar and solid contents decreased. When apple wine was fultered through $0.45\;{\mu}m$ nitrocellulose membrane followed by various ultrafiltration membranes with different molecular weight cut-off values, the initial flux $(121.2\;liter/m^2/h)$ and the average flux of Biomax 100k membrane were the highest among the membranes used. These membrane filtration treatments resulted in complete removal of microorganisms as well as decrease in turbidity and solid content without changes in other chemical properties. No changes in the physicochemical properties of the apple wine and no microorganisms were detected during the storage at $156{\circ}C$ for 6 weeks.

• The Korean Journal of Nuclear Medicine Technology
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• v.23 no.1
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• pp.64-68
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• 2019

Purpose In the preparation process for N-13 Ammonia injections, there were radioactive medicines adsorbed on filters remarkably. Hereby, we have compared the adsorption rate and quality test on Millex GS filter and Satorious Minisart filter, both representatively hydrophilic sterilizing filters, also evaluated which filter is more accommodative for N-13 Ammonia injection. Materials and Methods The filters used for sterilization of N-13 Ammonia injections were Millex GS filter($0.22{\mu}m$) mand Satorious Minisart filter ($0.2{\mu}m$), which are generally used to strain aqueous solutions. After the N-13 Ammonia passes through each sterilization filter, the adsorption rate of the filter (n=10) is determined by measuring not only the radioactivity through the filter also the amount of radioactivity remaining in it using a Dose Calibrator. The N-13 Ammonia injections after each filter is tested by the quality control test to conform to the Samsung Medical Center standard. Results The ratio of radioactivity passed through Millex GS indicated $29.0{\pm}17.6%$. Satorious Minisart filters output was $80.9{\pm}3.2%$, respectively. Each ratio of radioactivity adsorbed on the sterile filter was $71.0{\pm}17.6%$ for Millex GS and $19.1{\pm}3.2%$ for the Satorious Minisart filters, respectively. Furthermore, on the ratio of filtered radioactivity, Using Satorious Minisart filter showed about 2.8 times higher than using Millex GS filter. The quality testing of N-13 Ammonia injections through each filter met the Samsung Medical Center standard. Conclusion The Millex GS filter is composed of cellulose acetate and cellulose nitrate, whereas the Satorious Minisart filter if composed only of cellulose acetate. Therefore, the presence of cellulose nitrate in the membrane seems to have made differences. Therefore, the use of Satorious Minisart filter in the preparation of N-13 Ammonia injection solution minimized the loss of radioactive medicines due to filter adsorption, thereby improving the synthesis yield.

• Archives of Plastic Surgery
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• v.34 no.6
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• pp.679-684
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• 2007

Purpose: DMH(1,2-dimethylhydrazine) has been known to induce vascular neoplasm such as malignant endothelioma in animal experiment, through induction of abnormal proliferation of HUVECs. In our previous studies, 11 types of PKC isoenzymes were determined by RT-PCR and the expression of $PKC{\alpha}$, and ${\mu}$ was more prominent than other PKC isoenzymes in the DMH-treated group. However, this result was not based on objective assessment. In this study, we further evaluated the role of $PKC{\alpha}$ on the DMH-induced abnormal proliferation of HUVECs by two different methods to identify its presence with high relevance in objective view. $PKC{\mu}$ will be investigated in further study. Methods: The study was conducted with the cultured HUVECs group(control) and the $0.75{\times}10^{-9}M$ DMH-treated group. After processing protein extraction in 0 and 24 hour, extracted protein was treated of quantitative test through BCA protein assay. In the western blot analysis, electrophoresis was performed in the order of gel preparation, sample preparation, and gel running. Electrotransfer to nitrocellulose membrane and reaction with antibody were done. Detection of $PKC{\alpha}$ was achieved through "Gel Image Analysis System". In the fluorescence immunocytochemical analysis, the grading of radiance of the intracellular $PKC{\alpha}$ particles was detected with confocal microscope after treating with primary and fluorescent secondary antibody in 0 and 24 hours. Results: The Western blot analysis showed increased $PKC{\alpha}$ expression from the specimen obtained in 24 hour of the DMH treatment group when compared to those in control group. Under confocal fluorescence microscope, the emitting radiance in the DMH treated group was brighter at 24 hours as well. Conclusion: We believe that $PKC{\alpha}$ plays a role in DMH-induced abnormal proliferation of the vascular endothelium, which may provide insights in understanding the vascular neoplasm.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.436-439
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• 1989

The chromosomal DNA fragments of thermophilic alkalophilic Bacillus sp, K-17, a potent xylanhydrolyzing bacterium, were ligated to a vector plasmid pBR322 and transformed into Escherichia coli HB101. The plasmid pAX278, isolated from a transformant forming yellow color on the LB agar plate containing 1 mM p-nitrophenyl- $\beta$-xylopyranoside, was found to enable the transformants to produce p-xylosidase. The 5.0 kilobase insert of pAX278 had single sites for EcoRI, PstI, XbaI, and PvuII, and 2 sites for BglII. Biotinylated pAX218 was hybridized to 0.9 kb as well as 5.0 kb fragment from Bacillus sp. K-17 DNA on nitrocellulose filter. pGX718 was constructed by inserting the 5.0 kb HindIII fragment of pGX278 at the HindIII site of pGR71, E. coli and B. subtilis shuttle vector. The enzymatic properties of $\beta$-xylosidase from E. coli HB101 carrying recombinant plasmid were the same those of $\beta$-xylosidase from Bacillus sp. K-17.

• Journal of Conservation Science
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• v.25 no.4
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• pp.421-430
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• 2009

This research was carried out focusing on the urgent treatment and related studies for paint scribbling on Samjeondobi Monument (Historic Sites No. 101) in 2007. Before the preliminary test, the paint lacquer used on the surface of Samjeondobi Monument was analyzed. The paint lacquer turned out to be the paint lacquer spray composed of $Pb_3O_4$ used for the red pigment in the market. It was proved that the poultice used with the organic solvent was the best way to remove the paint pollutants following the preliminary test for the removal of paint pollutants which was performed with various removal methods by the laser, etc. However, the removing the paint pollutants was found in difficulty in contrast to the preliminary tests because the paint on the spot was hardened so rapidly over time that there was difference from the situation of the laboratory. For that problem, the poultice method with ethylene dichloride of main component from Remover (goods in the market) was the most efficient, therefore the pollutants were removed with the solution of alkyds resin and nitrocellulose and the rest part was removed by the $Laponite^{(R)}$ RD.

• Biomedical Science Letters
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• v.2 no.2
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• pp.175-185
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• 1996

The mitogen-activated protein(MAP) kinase signal transduction pathway represents an important mechanism by which mitogen, such as serum and PMA, regulate cell proliferation and differentiation. Target substrates of the MAP kinase are located within several compartments containing plasma membranes and nucleus. We now report that serum addition induces proliferation of the P388 murine leukemia cell, but PMA does not, while both serum and PMA treatment cause translocation of the MAP kinase, mainly p42$^{mapk}$ isoform, from cytosol into the nucleus, which was monitored by immunoblot analysis using polyclonal anti-ERK1 antibodies. We investigated whether the MAP kinase was capable of phosphorylating c-Jun protein and GST-fusion proteins, the P562$^{kk}$N-terminal peptides (1-77 or 1-123 domain) of the T cell tyrosine kinase, using the partially purified MAP kinase by SP-sephadex C-50, phenyl superose and Mono Q column chromatography. We found that the partially purified MAP kinase was able to phosphorylate c-Jun protein and the GST-fusion protein expressed using E.coli DH5$\alpha$ which is transformed with pGEX-3Xb plasmid vector carrying of p562$^{kk}$N-terminal peptide-encoding DNA. These results imply that tyrosine kinase receptor/Ras/Raf/MAP kinase pathway is a major mechanism for mitogen-induced cell proliferation in P388 murine leukemia cell and that the various MAP kinase isoforms may have their own target substrates located in distinct subcellular compartments.

• Parasites, Hosts and Diseases
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• v.26 no.3
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• pp.163-168
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• 1988

To observe the antigenic protein fractions in saline extract of Spirometra mansoni plerocercoid (sparganum), the crude extract was separated in reducing conditions of sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE). The proteins, transferred by celctrophoresis to introcillulose paper, were reacted with sera from 15 surgically confirmed sparganosis and 24 cysticercosis patients for immunoblotting. Out of 30 identified protein bands in the extract, bands of 29 and 36 kilodaltons (kDa) were the strongest and the most frequently reacting with specific antibody (IgG) in sparganosis sera. Bands of highter molecular weight also reacted with the sera but their frequency of reactions was lower. Sera of cysticercosis reacted with different protein bands in saline extract of sparganum, but the cross reactions were observed in strong antigenic bands of 29 and 36 kDa.

• Parasites, Hosts and Diseases
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• v.27 no.1
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• pp.9-14
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• 1989

Enzyme-linked immunoelectrotransfer blot (EITB) using crude worm antigen of adult Paragonimus westermani was performed for human patients sera to identify the species-specific components. Crude antigen was obtained by homogenizing and centrifuging 24-week old adult worms at 10,000 rpm for 60 minutes in phosphate buffered saline (PBS, PH 7.2) containing: Phenyl methyl sulfonyl auoride (PMSF). Gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) was performed and blotted electrophoretically onto a sheet of nitrocellulose paper. The sheet was cut into strips and exposed to sera diluted 1 : 200 with PBS. SDS-PAGE showed 26 protein bands ranging 229 to 10 kDa. Of them 229, 91, 60, 50, 35∼31, 27, 25, 21, 17, 11 and 10 kDa components showed positive reaction with serum antibody of patients with p. westermani. Sera of patients infected with Clcnorchis sinensis reacted with 35∼31, 19, and 11 kDa bands. Human sera from cysticercosis and diphyllobothriasis cases showed non-specific cross reactions with 229, 35∼31, 27, 25 and 17kDa bands. Protein bands of 91, 60, 21 and 10kDa showed strong positive reaction without cross reactions with sera from other helminthic infections.