• Korean Journal of Food Science and Technology
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• v.19 no.5
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• pp.397-402
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• 1987

Two lipoxygenases (F-I and F-II) were purified from potato tubers by ammonium sulfate fractionation and ion-exchange column chromatographies. The purified isoenzymes were apparently homogeneous on polyacrylamide gel electrophoresis. Both enzymes showed a similar optimum pH of 5.5-6.0. From thermal inactivation experiments with the purified enzymes in the range of 50 to $65^{\circ}C$, D-values of 13.3 min and 4.3 min at $65^{\circ}C$, and z-values of $11.8^{\circ}C\;and\;10.3^{\circ}C$ were obtained respectively for F-I and F-II. By applying absolute reaction rate equation, thermodynamic parameters wire also determined for the activation part of the inactivation process.

• KSBB Journal
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• v.25 no.5
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• pp.478-482
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• 2010

Inhibitory melanogenesis by Bambusae caulis in Teaniam (Phyllostaachys nigra var. henonis Stapf) was studied. Tyrosinase inhibition activities were evaluated with six different extracts. Among them the extract with methanol showed the highest tyrosinase activity inhibition. MTT assay with B16 melanoma showed that the extract was not toxic up to the concentration of 50 ppm. The melanogenesis was clearly inhibited by the extract when it was examined by the melanin content assay in the cell. When the extract was dosed as 10 ppm, the melanogenesis was reduced to 68% in culture medium and 74% in the cell. By the proteome analysis with 2-D electrophoresis, 171 protein spots were found in the control gel and 282 spots were detected in the sample gel. Among 120 spot proteins matched, 12 spots were identified as proteins involved in the melanogenesis mechanism.

• Biomedical Science Letters
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• v.11 no.3
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• pp.281-287
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• 2005

Although actinomycin D (AMD) is known to induce apoptotic cell death to various cell lines, the mechanism of apoptosis induced by AMD is still unclear. Understanding this mechanism may improve its therapeutic efficacy. The present study has been performed to elucidate expression of Bcl-2 and Caspase-3 proteins related to apoptosis in human leukemia K-562 cells. Five different assays were performed in this study; DNA fragmentation analysis by agarose gel electrophoresis, quantitative assay of fragmented DNA, morphological assessment of apoptotic cells, quantification of apoptosis by annexin V (AV) and propidium iodide (PI) staning, and expression of Bcl-2 and Caspase-3 proteins by the western blot analysis. The number of apoptotic cells and amount of fragmented DNA in this cell line treated with AMD was increased at 6 hour. DNA ladder pattern was also appeared at 6 hour. The expression of Bcl-2 was decreased, and disappeared from 12 hours after AMD treatment. Precursor of Caspase-3 was degraded, and 20 kDa cleavage products were detected. These results suggest that AMD induced apoptosis of K-562 cells is Caspase-3-dependent fashion, and this apoptosis is related to the degradation of Bcl-2 proteins.

• Journal of the Korean Society of Industry Convergence
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• v.14 no.3
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• pp.105-111
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• 2011

Landmark Spots in 2D gel electrophoresis are used in many methods of 2DEG spot matching. Landmark Spots are obtained manually and it is a bottle neck in the entire protein analysis process. Automated landmark spots detection is a very crucial topic in processing a massive amount of 2DGE data. In this paper, Automated landmark spot detection is proposed using point pattern matching and graph theory. Neighbor spots are defined by a graph theory to use and only a centered spot and its neighbor spots are considered for spot matching. Normalized Hausdorff distance is introduced as a criterion for measuring degree of similarity. In the conclusion, the method proposed in this paper can get about 50% of the total spot pairs and the accuracy rate is almost 100%, which the requirements of landmark spots are fully satisfied.

• Journal of Microbiology and Biotechnology
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• v.10 no.1
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• pp.27-34
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• 2000

The chloroform and methanol (2;1, v/v) extract from an edible plant, Actinidia arguta Planchon, appeared to possess antitumor activity against human leukemias Jurkat T and U937 cells through inducing apoptosis. The substance in the solvent extract was purified by silica gel column chromatography, preparative TLC, and Sephadex LH-20 column chromatography. Characteristics of the substance analyzed by UV scanning analysis, $^1H$ and $^{13}C$ NMR spectra suggested that the substance belongs to the chlorophyll derivatives-like group. The $IC_{50}$ value of the chlorophyll derivative (Cp-D) determined by MTT assay was $15\mu\textrm{g}/ml$ for Jurkat, $10\mu\textrm{g}/ml$ for U937, and $11.4\mu\textrm{g}/ml$ for HL-60m and was more toxic to these leukemias than to solid tumors or normal fibroblast. In order to elucidate cellular mechanisms underlying the cytotoxicity, the effect of the Cp-D on Jurkat T cells was investigated. When cells were treated with the Cp-D at a concentration of $15\mu\textrm{g}/ml$, [3H]thymidine incorporation declined rapidly and wa undetectable in 1h. However, no significant changes were made in the cell cycle distribution of the cells by 24h. The sub-Gl peak representing apoptotic cells began to be detectable in 36h, at which time apoptotic DNA fragmentation was also detected on agarose gel electrophoresis, demonstrating that the cytotoxic effect of the Cp-D is attributable to the induced apoptosis. Under the same conditions, although the protein level of cyclin-dependent kinases such as cdc4, csk6, cdk2, and cdc2 was not significantly changed until 24h, the kinase activity of all c안 rapidly declined and reached a minimum level within 1-6h and then recovered to the initial level by 12h and sustained until 24h. These results suggest that inactivation of cdks at an inappropriate time during the cell cycle progression in jurkat T cells following a treatment with the Cp-D leads to induction of apoptotic cell death.

• Journal of Microbiology and Biotechnology
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• v.22 no.10
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• pp.1330-1335
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• 2012

Foot and mouth disease (FMD) is one of the acute infectious diseases in hoofed and even-toed mammals, including pigs, and it occurs via acute infection by Aphthovirus. When FMD is suspected, animals around the location of origin are typically slaughtered and buried. Other methods such as rendering, composting, and incineration have not been verified in practice in Korea. After the FMD incident, the regular monitoring of the microbial community is required, as microorganisms greatly modify the characteristics of the ecosystem in which they live. This is the result of their metabolic activities causing chemical changes to take place in the surrounding environment. In this study, we investigated changes in the microbial community during a 24 week period with DNA extracts from leachate, formed by the decomposition of buried pigs at a laboratory test site, using denaturing gradient gel electrophoresis (DGGE) with a genomic DNA. Our results revealed that Bacteroides coprosuis, which is common in pig excreta, and Sporanaerobacter acetigenes, which is a sulfur-reduced microbe, were continuously observed. During the early stages (0~2 weeks) of tissue decomposition, Clostridium cochlearium, Fusobacterium ulcerans, and Fusobacterium sp., which are involved in skin decomposition, were also observed. In addition, various microbes such as Turicibacter sanguinis, Clostridium haemolyticum, Bacteroides propionicifaciens, and Comamonas sp. were seen during the later stages (16~24 weeks). In particular, the number of existing microbial species gradually increased during the early stages, including the exponential phase, decreased during the middle stages, and then increased again during the later stages. Therefore, these results indicate that the decomposition of pigs continues for a long period of time and leachate is created continuously during this process. It is known that leachate can easily flow into the neighboring environment, so a long-term management plan is needed in burial locations for FMD-infected animals.

• Environmental Mutagens and Carcinogens
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• v.26 no.2
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• pp.35-40
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• 2006

Background: Inorganic arsenic is a human carcinogen that can target the liver, but its carcinogenic mechanisms are still unknown. Inorganic arsenic induces a spectrum of tumors including hepatocellular carcinoma in mice. Methods: Pregnant C3H mice were supplied with drinking water containing 50 ppm sodium arsenite during their pregnancy. The protein expression profile in the liver of 0.5-day-old. male offsprings exposed transplacentally to sodium arsenite was analyzed using protein 2D gel electrophoresis followed by mass spectrometry (MALDI-TOF). Results: Expression of proteins such as hydroxymethylglutaryl-CoA synthase mitochondrial precursor (HMG-CoA synthase), ${\beta}$-actin (cytoplasmic 1) and apolipoprotein A-IV precursor (Apo-AIV) were induced in mouse liver by sodium arsenite, while uricase (urate oxidase), guanine nucleotidebinding protein beta subunit 2-like 1 (RACK1) and fructose-bisphosphate aldolase B (Aldolase 2) were down-regulated. Summary: Expression of proteins that have been implicated in carcinogenesis, such as HMG-CoA, ${\beta}$-actin, and RACK1, was regulated in the liver of mice transplacentally exposed to inorganic arsenic.

• Journal of Veterinary Clinics
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• v.25 no.3
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• pp.152-158
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• 2008

We investigated the changes of protein in chronic MI which was occurred with long-term ischemia, without reperfusion. Sprague Dawley (SD) rats were divided into the sham group and the experimental groups (MI groups). The sham group was treated only thoracotomy without ligation for left main descending artery (LMDA) of left coronary artery (LCA), and the experimental groups (MI7d, ligation of LMDA for 7 days and MI30d, ligation of LMDA for 30 days) were conducted an artificial chronic MI. The change of proteins according to passage of times was compared and analyzed on first and second dimension (1 and 2D) sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Among total 46 spots expressed differentially in the sham group versus MI7d and MI30d groups on 2D gel, we selected proteins that the volume of spot was increased in the MI7d and MI30d groups compared with the sham group. After that, the proteins were identified through liquid chromatography/tandem mass spectrometry (LC-MS/MS) analysis. In result, we could obtain many proteins as follows; albumin, glucose regulated protein 58 KDa, similar to tripartite motif protein 50, ubiquinol-cytochrome c reductase core protein II, sarcomeric mitochondrial creatine kinase, ATP synthetase alpha chain (mitochondrial precursor) and creatine kinase. In conclusion, we suggest many changed proteins shown at chronic ischemia after artificial MI and consider that these proteins play an important role in the function of heart after MI.

• The Korean Journal of Zoology
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• v.28 no.1
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• pp.21-30
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• 1985

By two-dimensional gel electrophoresis loss of a cell-specific protein was detected in tD strain of Amoeba proteus that had been infected by symbiotic bacteria extracted from xD strain. In 50 days of experimental infection by induced phagocytosis the host amoeba lost the ability to synthesize the tD cell-specific protein even after removal of the infective bacteria and xD cell-specific protein by growing the amoebae at $27^\\circC$. By this time the host amoebae were obligately dependent on the bacteria. From these and other results (Lorch and Jeon, Science 221:549), it is clear that the incompatibility of the infected nuclei with the cytoplasm of the uninfected amoeba and the obligate dependence of the host on bacteria are due to the irreversible inactivation or the loss of the cell-specific gene by bacterial infection in this amoeba.

• Toxicological Research
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• v.13 no.1_2
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• pp.117-125
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• 1997

In order to assess the possibility whether CYP2D is involved in caffeine metabolism, we have purified and characterized the rat liver microsomal cytochrome P4502D1 (CYP2D1), equivalent to CYP2D6 in human liver, and have utilized the reconstituted CYP2D1 in the metabolism of 4 primary caffeine (1, 3, 7-trimethylxanthine) metabolites such as paraxanthine (1, 7-dimethylxanthine), 1, 3, 7-trimethylurate, theophylline (1, 3-dimethylxanthine) and theobromine (3, 7-dimethylxanthine). Rat liver CYP 2D1 has been purified to a specific content of 8.98 nmole/mg protein (13.4fold purification, 1.5% yield) using $\omega$-aminooctylagarose, hydroxlapatite, and DE52 columns in a sequential manner. As judged from sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), the purified CYP2D1 was apparently homogeneous. Molecular weight of the purified CYP2D1 was found to be 51, 000 Da. Catalytic activity of the purified and then reconstituted CYP2D1 was confirmed by using bufuralol, a known subsFate of CYP2D1. The reconstituted CYP2D1 was found to produce to 1-hydroxylbufuralol at a rate of 1.43$\pm$0.13 nmol/min/nmol P450. The kinetic analysis of bufuralol hydroxylation indicated that Km and Vmax values were 7.32$\mu M$ and 1.64 nmol/min/nmol P450, respectively. The reconstituted CYP2D1 could catalyze the 7-demethylation of PX to 1-methylxanthine at a rate of 12.5 pmol/min/pmol, and also the 7- and 3- demethylations of 1, 3, 7-trimethylurate to 1, 3-dimethylurate and 1, 7-dimethylurate at 6.5 and 12.8 pmol/min/pmol CYP2D1, respectively. The reconstituted CYP2D1 could also 3-demethylate theophylline to 1-methylxanthine at 5 pmol/min/pmol and hydroxylate the theophylline to 1, 3-dimethylurate at 21.8 pmol/min/pmol CYP2D1. The reconstituted CYP2D1, however, did not metabolize TB at all (detection limits were 0.03 pmol/min/pmol). This study indicated that CYP2D1 is involved in 3-and 7-demethylations of paraxanthine and theophylline and suggested that CYP2D6 (equivalent to CYP2D1 in rat liver) present in human liver may be involved in the secondary metabolism of the primary metabolites of caffeine.