• KSBB Journal
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• v.26 no.6
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• pp.517-522
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• 2011

In our previous study, we isolated and characterized a 1-deoxynojirimycin (DNJ)-producing bacterium, Bacillus subtilis MORI, from chungkookjang, a Korean traditional food. B. subtilis MORI was subjected to ${\gamma}$-irradiation and the resulting bacteria were screened for increased DNJ production. A mutant was identified that produced 7.6 times more DNJ and named B. subtilis MORI 3K-85. In this study, the protein profiles of both strains were compared by one-dimensional and two-dimensional gel electrophoresis (1-DE and 2-DE, respectively) under both native and denaturing conditions. The 1-DE native-PAGE and 1-DE SDS-PAGE analyses identified 5 and 7 bands, respectively, that were found at higher concentrations in B. subtilis MORI 3K-85 than in B. subtilis MORI. Similarly, 2-DE analyses identified 20 protein spots which were found at higher concentrations in B. subtilis MORI 3K-85. The peptide mass profiles of these 20 proteins were analyzed by MALDI-TOF and compared with peptide sequences of B. subtilis and B. amyloliquefaciens in the MASCOT database. This screening suggested that three dehydrogenases, an aldolase, a synthetase, an isomerase, a reductase, and a peroxidase are elevated in B. subtilis MORI 3K-85. Based on this data, one or more of the elevated 8 enzymes might be related to the DNJ biosynthetic pathway.

• Molecular & Cellular Toxicology
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• v.1 no.2
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• pp.116-123
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• 2005

The application of high pressure on cellular morphology, proliferation and protein expression of Jurkat cells (human T lymphocyte cell line) has been extensively investigated. In the present study, we manufactured a novel pressure chamber that modulates 5% $CO_{2}$, temperature and pressure (up to 3 ATA). Jurkat cells was incubated 2 ATA pressure and analyzed cellular morphology and growth using an electron microscopy and MTT assay. The cells showed the morphological changes in the cell surface, which appeared to cause a severe damage in cell membrane. The growth rate of the cells under 2 ATA pressure decreased as cultured time got increased. Furthermore, a long term exposure of high pressure on Jurkat cells may act as one of the important cellular stresses that leads to inducing cell death. Cellular proteomes were separated by 2-dimensional electrophoresis with pH 3-10 ranges of IPG Dry strips. And many proteins showed significant up-and-down expressions with hyperbaric pressure. Out of all, 10 spots were identified significantly using matrix-assisted laser desorption/ionization-time of fight (MALDI-TOF) mass spectrometry. We and found that 9 protein expressions were decreased and one protein, heat shock protein HSP 60, was increased in Jurkat cells under 2 ATA. Identified proteins were related to lipid metabolism and signal transduction.

• Proceedings of the Korea Society of Poultry Science Conference
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• 2005.11a
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• pp.66-67
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• 2005

The domestic chicken (Gallus gallus) is an important model for research in developmental biology because its embryonic development occurs in ovo. To examine the mechanism of embryonic germ cell development, we constructed proteome map of gonadal primordial germ cells (gPGC) from chicken embryonic gonads. Embryonic gonads were collected from 500 embryos at 6 day of incubation, and the gPGC were cultured in vitro until colony formed. After 7-10 days in cultured gPGC colonies were separated from gonadal stroma cells (GSCs). Soluble extracts of cultured gPGCs were then fractionated by two-dimensional gel electrophoresis (pH 4-7). A number of protein spots, including those that displayed significant expression levels, were then identified by use of matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry and LC-MS/MS. Of the 89 gPGC spots examined, 50 yielded mass spectra that matched avian proteins found in on-line databases. Proteome map of thistype will serve as an important reference for germ cell biology and transgenic research.

• Journal of Life Science
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• v.14 no.5
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• pp.847-854
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• 2004

Allergy is a multi-factorial disease influenced by genetic and environmental factors. As the number of allergy-affected people is increasing in developed countries, there is an increasing interest in genetic predisposition to the allergy. A number of genes and chromosomal region have been identified to be linked to allergy including rhinitis, asthma and atopy. In order to understand the genetic background for the allergy-affected people, we investigated genetic predisposition among students enrolled in Busan Science Academy. Among 138 students, about 30% students had some allergy-related disorder including rhinitis, asthma and atopy. We analyzed several single nucleotide polymorphisms (SNPs) within two genes, Inter-leukin-4(IL-4) and Interleukin-4 receptor(IL-4R), which are involved in the induction of allergy reaction with the Th2 immunity. For 96 samples obtained from students, we analyzed 9 SNPs including -590 C/T and -34 C/T in IL-4, and I75V, Q576R, E375A, e406R, 5411L, S761P and S727A in IL-4R. From the analysis, these SNPs showed slight differences among normal and allergy-affected students, but these differences was not enough to predict the predisposition to the allergy. In contrast to previous reports, we could not find SNP(s) related with allergy. These results suggest that genetic tests recently performed in Korea widely have to be reassessed for its validity of genetic predisposition. [Supported by grants from MOST]

• Archives of Pharmacal Research
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• v.29 no.3
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• pp.224-234
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• 2006

We employed human SK-MEL-28 cells as a model system to identify cellular proteins that accompany N-(4-methyl)phenyl-O-(4-methoxy)phenyl-thionocarbamate (MMTC)-induced apoptosis based on a proteomic approach. Cell viability tests revealed that SK-MEL-28 skin cancer cells underwent more cell death than normal HaCaT cells in a dose-dependent manner after treatment with MMTC. Two-dimensional electrophoresis in conjunction with matrixassisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry analysis or computer matching with a protein database further revealed that the MMTC-induced apoptosis is accompanied by increased levels of caspase-1, checkpoint suppressor-1, caspase-4, NF-kB inhibitor, AP-2, c-Jun-N-terminal kinase, melanoma inhibitor, granzyme K, G1/S specific cyclin D3, cystein rich protein, Ras-related protein Rab-37 or Ras-related protein Rab-13, and reduced levels of EMS (oncogene), ATP synthase, tyrosine-phosphatase, Cdc25c, 14-3-3 protein or specific structure of nuclear receptor. The migration suppressing effect of MMTC on SK-MEL-28 cell was tested. MMTC suppressed the metastasis of SK-MEL-8 cells. It was also identified that MMTC had little angiogenic effect because it did not suppress the proliferation of HUVEC cell line. These results suggest that MMTC is a novel chemotherapeutic and metastatic agents against the SK-MEL-28 human melanoma cell line.

• Korean Journal of Clinical Laboratory Science
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• v.52 no.3
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• pp.214-220
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• 2020

Handwashing with soap is an important practice to reduce the transmission of potentially pathogenic microorganisms, but liquid soaps with refillable dispensers are prone to extrinsic bacterial contamination. This study investigated the bacterial contamination of liquid soaps in 58 public restrooms in six buildings. The bacteria were identified by a biochemical test and MALDI-TOF mass spectrophotometry. The antimicrobial susceptibility was determined using the Vitek II system. Of the 58 restrooms examined, 27(46.55%) were using a refill dispenser, of which 25(92.59%) were contaminated with bacteria. The bacteria recovered from the soaps ranged from 1.6×10³ to 2.7×10⁵ CFU/mL. Serratia liquefaciens (12), Achromobacter xylosoxidans (9), S. marcescens (4), Staphylococcus pastueri (1), and Achromobacter spanius (1) were isolated. Except for one A. xylosoxidans, bacteria of the same species isolated in the same building showed a unique resistance pattern. In conclusion, handwashing with contaminated soap may play a role in the transmission of bacteria in public health settings. Therefore, it is necessary to limit the use of refillable liquid soaps in the restrooms of hospitals used by patients with reduced immunity.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.9
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• pp.1239-1246
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• 2016

The breeding of yaks is highly seasonal, there are many crucial proteins involved in the reproduction control program, especially in follicular development. In order to isolate differential proteins between mature and immature follicular fluid (FF) of yak, the FF from yak follicles with different sizes were sampled respectively, and two-dimensional gel electrophoresis (2-DE) of the proteins was carried out. After silver staining, the Image Master 2D platinum software was used for protein analysis and matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was performed for differential protein identification. The expression level of transferrin and enolase superfamily member 1 (ENOSF1) was determined by Western blotting for verification analysis. The results showed that 2-DE obtained an electrophoresis map of proteins from mature and immature yak FF with high resolution and repeatability. A comparison of protein profiles identified 12 differently expressed proteins, out of which 10 of them were upregulated while 2 were downregulated. Western blotting showed that the expression of transferrin and ENOSF1 was enhanced with follicular development. Both the obtained protein profiles and the differently expressed proteins identified in this study provided experimental data related to follicular development during yak breeding seasons. This study also laid the foundation for understanding the microenvironment during oocyte development.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.11
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• pp.1540-1545
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• 2012

Steroidogenesis requires coordination of the anabolic and catabolic pathways of lipid metabolism, but the profile of proteins associated with progesterone synthesis in cyclic and pregnant corpus luteum (CL) is not well-known in cattle. In Experiment 1, plasma progesterone level was monitored in cyclic cows (n = 5) and pregnant cows (n = 6; until d-90). A significant decline in the plasma progesterone level occurred at d-19 of cyclic cows. Progesterone level in abbatoir-derived luteal tissues was also determined at d 1 to 5, 6 to 13 and 14 to 20 of cyclic cows, and d-60 and -90 of pregnant cows (n = 5 each). Progesterone level in d-60 CL was not different from those in d 6 to 13 CL and d-90 CL, although the difference between d 6 to 13 and d-90 was significant. In Experiment 2, protein expression pattern in CL at d-90 (n = 4) was compared with that in CL of cyclic cows at d 6 to 13 (n = 5). Significant changes in the level of protein expression were detected in 32 protein spots by two-dimensional polyacrylamide gel electrophoresis (2-DE), and 23 of them were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Six proteins were found only in pregnant CL, while the other 17 proteins were found only in cyclic CL. Among the above 6 proteins, vimentin which is involved in the regulation of post-implantation development was included. Thus, the protein expression pattern in CL was disorientated from cyclic luteal phase to mid pregnancy, and alterations in specific CL protein expression may contribute to the maintenance of pregnancy in Korean native cows.

• Herbal Formula Science
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• v.24 no.1
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• pp.17-30
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• 2016

Objectives Rehmanniae Radix Preparata was prepared in the traditional Rehmanniae Radix Preparata preparation method set forth in the 『Treasured Mirror of Eastern Medicine(Donguibogam)』 with a view to measuring the contents of 5-Hydroxymethyl-2-furaldehyde(5-HMF) at individual stages of steaming and sundrying and identifying new chemical components.Methods Based on the traditional Rehmanniae Radix Preparata preparation method set forth in the 『Treasured Mirror of Eastern Medicine』, Rehmanniae Radix Preparata steamed and sundried once through nine times was prepared. Thereafter, 5-HMF contents were analyzed and new chemical components were identified in the Rehmanniae Radix Preparata using Waters HPLC e2695, 2640 detectors, a Waters Acquity UPLC system, and a Micromass Q-TOF Premier mass spectrometer.Results The Rehmanniae Radix Preparata preparation method set forth in the 『Treasured Mirror of Eastern Medicine』 is a unique preparation method in Republic of Korea different from that in China. In the first stage of the method, fresh Rehmanniae Radix Crudus was divided into high quality, medium quality, and low quality ones named Rehmanniae Radix Crudus (Caelum)(天黃), Rehmanniae Radix Crudus (Homo)(人黃), and Rehmanniae Radix Crudus (Terra)(地黃) respectively to use Rehmanniae Radix Crudus (Caelum) and Rehmanniae Radix Crudus (Homo) for preparation of juice while using Rehmanniae Radix Crudus (Terra) to make Rehmanniae Radix Preparata. In the second stage, Rehmanniae Radix Crudus (Caelum) and Rehmanniae Radix Crudus (Terra) were made into juice and Rehmanniae Radix Crudus (Terra) was soaked in the juice. In the third stage, among auxiliary materials, rice wine named Purum Vinum Oryzae(淸酒) brewed from sticky rice was sprinkled on Rehmanniae Radix Crudus (Terra) to the extent that Rehmanniae Radix Crudus (Terra) became wet. In the fourth stage, Rehmanniae Radix Preparata steamed in earthenware steamer was dried under natural sunlight. The contents of 5-HMF in Rehmanniae Radix Preparata steamed and sundried once through nine times were shown to be below 0.1% in all cases. Pomolic acid was identified as a new chemical component.Conclusions In conclusion, the Rehmanniae Radix Preparata preparation method set forth in the 『Treasured Mirror of Eastern Medicine』 is thought to be a unique preparation method in South Korea in which Rehmanniae Radix Preparata is completed through the first stage in which fresh Rehmanniae Radix Crudus collected from fields is divided into high, medium, and low quality ones and fresh Rehmanniae Radix Crudus juice is made, the second stage in which the high quality fresh Rehmanniae Radix Crudus is soaked in the fresh Rehmanniae Radix Crudus juice, the third stage in which the fresh Rehmanniae Radix Crudus is steamed, and the fourth stage in which the steamed Rehmanniae Radix Crudus is dried.

• Journal of the Korean Applied Science and Technology
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• v.31 no.2
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• pp.320-328
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• 2014

Biodiesel has very similar physical properties (density, kinematic viscosity) and has even higher cetane number compare with conventional diesel. There are no necessity to change or modify the infra-structure & engine system. It is known that fatty acid methyl ester (FAME) is oxygen-contained components increasing the combustibility, biodegradability and reduced the exhaust harmful gas. These things made the biodiesel more popular as an alternative diesel fuel. But biodiesel's sources are controversial issues about $CO_2$ reduction effect at this time because those mainly come from edible plants such as soy, palm, rapeseed already spent lot of $CO_2$ to cultivate. Whereas micro-algae is focused because they are inedible and has rapid growth rates & high carbon-dioxide adsorption rate per area. In this study, we analyze the each FAME components using $GC{\times}GC$-TOFMS in stead of GC-FID and verify the previous total FAME contents method's applicability through the micro algal biodiesel derived from Dunaliella tertiolecta.