• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.69-69
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• 2003

There are now increasing evidences that free radicals and reactive oxygen species are involved in a variety of pathological events. Reactive Oxygen Species (ROS) are produced during normal cellular function. ROS lead to lipid peroxidation, massive protein oxdiation and degradation. Under normal conditions, antioxidant are substnaces that either directly or indirectly protect cell against adverse effect of ROS. several biologically important compound include ${\beta}$-carotene, taruine and flavonoids reported have antioxidant function. The various antioxidant either scavange superoxide and free radicals or stimulate the detoxification mechanisms within cells resulting in increased detoxification of free radicals formation and thus in prevention of many pathophysiologic processes. This study carried out to investigate the antioxidant activity of flavonoids, myricetin with other antioxidants, ${\beta}$-carotene and taurine on B16Fl0. In order to investigate the efficacy of antioxidant activity, we measured cell viability, antioxidant enzyme activity (SOD, GPX, CAT) and intracellular reactive oxygen intermediate (ROI). In this results, we show that these flavonoids with other antioxidant substrates are increased antioxidant activity level.

• Bulletin of the Korean Chemical Society
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• v.20 no.9
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• pp.1073-1077
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• 1999

CRAMP, a 37-amino acid cationic antimicrobial peptide was recently deduced from the cDNA cloned from mouse femoral marrow RNA. In order to investigate the structure-activity relationship and functional region of CRAMP, CRAMP and its 18-mer overlapping peptides were synthesized by the solid phase method. CRAMP showed broad spectrum antibacterial activity against both Gram-positive and Gram-negative bacterial strains (MIC: 3.125-6.25 μM) but had no hemolytic activity until 50 μM. CRAMP was found to have a potent anticancer activity (IC50: 12-23 μM) against two human small cell lung cancer cell lines. Furthermore, CRAMP was found to display faster bactericidal rate in B. subtilis rather than E. coli in the kinetics of bacterial killing. Among 18-meric overlapping fragment peptides, only CRAMP (16-33) displayed potent antibacterial activity (MIC: 12.5-50 μM) against several bacteria with no hemolytic activity. Circular dichroism (CD) spectra anal-ysis indicated that CRAMP and its analogues will form the amphipathic α-helical conformation in the cell membranes similar to other antimicrobial peptides, such as cecropins and magainins.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.313-317
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• 2011

This study was conducted to examine the effect of oocyte donor age and micromanipulation medium on the development of mouse cloned embryos receiving cumulus cells. Mouse oocytes were obtained from 6 to 11 week-old mice BDF1 female mice(experiment 1) and cumulus cells were used as donor cells. Micromanipulation procedures for nuclear transfer(NT) were performed in FHM, M2 or Hepes-buffered TCM199(TCM199) medium(experiment 2). After nuclear transfer, the reconstructed oocytes were activated by 10 mM $SrCl_2$ in Ca-free CZB medium in the presence of 5 II ${\mu}$g/ml cytochalasin B for 5 h and cultured in KSOM medium for 4 days. In experiment 1, the survival rate of oocytes after injection of cumulus cells were significantly(p<0.05) lower in oocytes from 6~7 week-old mice(53.3%) than in oocytes from 8~9(80.9%) and 10~11 week-old mice(77.1%). In experiment 2, the survival rate of oocytes after cell injection were significantly(p<0.05) higher in FHM and M2 medium(71.7% and 76.9%) than in TCM199 medium(51.2%). The activation rates of cloned embryos were not different among the micromanipulation media. However, the embryos developed to blastocyst stage were significantly(p<0.05) higher in FHM medium(13.9%) than in M2 and TCM199 medium(0.0% and 0.0%). In conclusion, the present study suggest that oocytes from above 8 week-old mice are superior to oocytes from 6~7 week-old mice as a source of recipient cytoplasm and FHM is superior to M2 and TCM199 as a micromanipulation medium for mouse somatic cell cloning.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.145-152
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• 2007

Transgenic mice were generated by microinjecting a plasmid DNA containing the SV40 (simian virus 40) large T antigen (Tag) gene fused with mouse albumin promoter/enhancer sequences into fertilized one-cell mouse embryos. Among eleven founder transgenic animals, four developed hepatocellular carcinoma, two showed kidney cancer and one developed skin and brain tumors. Three stable transgenic lines, #1-2, #1-6 and #1-11 were established. Members of the lines #1-6 and #1-11 reproducibly developed liver tumors by 8 to 10 weeks of age but did not exhibit any phenotypic changes in other tissues. Histological changes loading to liver tumor formation occurred with predictable kinetics and could be classified into three distinct stages; (a) newborn to 3 weeks of age, characterized by hyperplastic hepatocytes with reduced amounts of cytoplasm without any nuclear alterations, (b) between 4 to 8 weeks of age, characterized by diffuse liver cell dysplasia without observable tumor nodules, and (c) 9 weeks of age and thereafter, characterized by hepatocellular carcinomas in the background of extensive liver dysplasia. Metastasis to the lung from a liver carcinoma was observed in #1-11 founder animal. This transgenic mouse system displays similarities with human liver cancers in a number of aspects and provides a useful model for the study of molecular events involved in hepatocarcinogenesis.

• Korean Journal of Food Science and Technology
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• v.51 no.2
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• pp.152-159
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• 2019

To characterize new physiologically active components in Korean apple vinegar, a crude polysaccharide (KAV-0) was prepared by precipitation with 80% (v/v) ethanol. KAV-0 mainly comprises 38.2% mannose, 19.1% galactose and 14.3% glucose. In an in vitro cytotoxicity analysis, KAV-0 promoted the proliferation of peritoneal macrophages and RAW 264.7 cells in a dose-dependent manner, and showed no cytotoxicity in B16-BL6 melanoma cells. Murine peritoneal macrophages and RAW 264.7 cells stimulated by KAV-0 produced various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factor $(TNF)-{\alpha}$, and nitric oxide (NO). Intravenous (i.v.) administration of KAV-0 significantly augmented NK cell cytotoxicity against Yac-1 tumor cells. In experimental lung metastasis caused by B16-BL6 melanomas, prophylactic i.v. administration of KAV-0 at a dosage of $1,000{\mu}g/mouse$ inhibited lung metastasis by 53.0%. These results suggest that the crude polysaccharide (KAV-0) isolated from Korean apple vinegar has a considerably high anti-metastatic activity and immunomodulatory activities beneficial to human health.

• Molecular & Cellular Toxicology
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• v.2 no.2
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• pp.81-88
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• 2006

Sphingolipid metabolites regulate many aspects of cell proliferation, differentiation, and apoptosis. In the present study, we have assessed the effects of the novel phytosphingosine derivative, N-acetylphytospingosine (NAPS), on the depigmentation of murine B16F10 melanoma cells, and have also attempted to identify the possible signaling pathway involved, in comparison with $C_{2}-ceramide$. NAPS and $C_{2}-ceramide$ both inhibited the growth of the B16F10 cells in a dose-dependent manner. Melanin content and tyrosinase activity were significantly reduced in response to treatment with NAPS and $C_{2}-ceramide$ at concentrations in a range between $1-5\;{\mu}M$. However, the levels of tyrosinase mRNA, as well as the levels of tyrosinase related protein-1 (TRP-1) and tyrosinase related protein-2 (TRP-2) genes and the level of tyrosinase protein remained unaffected by treatment with either NAPS or $C_{2}-ceramide$. We also attempted to determine the signaling pathway exploited by NAPS and $C_{2}-ceramide$. Interestingly, the phosphorylation of Akt/PKB at serine 473 by NAPS was reduced at the 5 minute mark, whereas $C_{2}-ceramide$ induced the phosphorylation of Akt/PKB at serine 473. Finally, Akt/PKB activity in the NAPS-treated cells was elevated in comparison with the untreated cells. LY294002, a specific PI3-K inhibitor which is located upstream of Akt/PKB, inhibited the phosphorylation of Akt/PKB, but induced an increase in melanin synthesis. These results suggest that the activation of Akt/PKB at serine 473 is related with the suppression of melanin production in the B16F10 mouse melanoma cells. Therefore, the mechanisms exploited by NAPS and $C_{2}-ceramide$ responsible for the depigmentation of B16F10 cells were concluded to involve the inhibition of melanosomal tyrosinase activity.

• Archives of Pharmacal Research
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• v.28 no.12
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• pp.1350-1357
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• 2005

Nuclear factor kappa B (NF-$\kappa$B) regulates the expression of multiple cytokines, chemokines, and cell adhesion molecules that are involved in the pathogenesis of asthma. We investigated the anti-asthmatic effects and the mechanism of action of DA-9201, an extract of the black rice, in a mouse model of asthma. Mice immunized with ovalbumin (OVA) were administered with DA-9201 (30, 100 or 300 mg/kg) or dexamethasone (DEXA, 3 mg/kg) for 2 weeks and challenged with aerosolized OVA during the last 3 days. Anti-asthmatic effects were assessed by means of enhanced pauses, level of total lgE and Th2 cytokines in plasma or bronchoalveolar lavage fluid (BALF), the percentage of eosinophils in BALF, and histopathological examination. The expression of NF-$\kappa$B in nuclear and cytoplasmic fraction and its DNA-binding activity in lung tissues were analyzed by means of Western blotting and electrophoretic gel mobility shift assay (EMSA), respectively. DA-9201 significantly reduced airway hyperrespon-siveness (AHR), total lgE level in plasma and BALF, IL-4, IL-5, and IL-13 levels in BALF, and the percentage of eosinophils in BALF. Tissue inflammation was significantly improved by DA­9201 treatment. In addition, DA-9201 dramatically suppressed the expression of NF-$\kappa$B and its DNA-binding activity. These results suggest that DA-9201 may be useful for the treatment of asthma and its efficacy is related to suppression of NF-$\kappa$B pathway.

• Korean Journal of Veterinary Research
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• v.39 no.2
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• pp.301-310
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• 1999

Anthrax caused by Bacillus anthracis is one of the most important zoonotic diseases. The bacterium produces several virulence factors. Of the factors, protective antigen (PA) of tripatite toxin has been identified as a central component in the pathogenesis of anthrax. However, precise roles of PA and other cellular components in the reaction with the target cells remain to be elucidated, especially in the initial stage of the disease. Three B anthracis antigens were prepared for investigation; PA, sonicated cellular antigens (S-Ag) and formalin-inactivaed whole cell antigens (W-Ag). PA was purified from culture supernatant of the bacterium using FPLC system with MonoQ. S-Ag and W-Ag were prepared by sonication and formalin inactivation of the cultured cells, respectively. Purity of the antigens was confirmed by SDS-PAGE and Western blot analysis. The roles of these antigens in the production of inflammatory mediators such as NO, IL-6 and $TNF{\alpha}$ from mouse peritoneal macrophages were investigated. PA alone did not induce the production of the inflammatory mediators while the other antigens, S-Ag and W-Ag, did in a dose and time dependent manner. These results suggested that in addition to major virulence factors, other cellular antigens are also involved in the initial stage of the disease by the induction of inflammatory mediators.

• The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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• v.16 no.2
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• pp.96-118
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• 2003

The effect of Glycyrrhizae Radix water extract, known as depigmenting agent, on melanin biosynthesis was investigated in cellular level by using B16 mouse melanoma cells. The inhibitory effect of Glycyrrhizae Radix water extract on melanogenesis was determined by mushroom tyrosinase assay traditionally using in vitro screening test. To determine whether Glycyrrhizae Radix water extract suppress melanin synthesis in cellular level, B16 mouse melanoma cells were cultured in the presence of different concentrations of Glycyrrhizae Radix water extract. Effects on cell proliferation, melanin biosynthesis, tyrosinase activity, DOPAchrome tautomerase activity, and expression level of mRNA for tyrosinase were examined. The maximum concentration of Glycyrrhizae Radix water extract that was not inhibitory to growth of the cells was 2 mgml. At that concentration, melanin synthesis was significantly inhibited without cytotoxicity after 5 days, compared with untreated cells. The treatment with Glycyrrhizae Radix water extract reduced tyrosinase and DOPAchrome tautomerase activity in a dose-dependent manner. However, the treatment with Glycyrrhizae Radix water extract did not affect significantly mRNA levels for tyrosinase. These results suggest that the inhibitory effect of Glycyrrhizae Radix water extract on melanogenesis is correlated with the suppression of tyrosinase and DOPAchrome tautomerase activity more than altering mRNA levels of tyrosinase.

• International Journal of Industrial Entomology and Biomaterials
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• v.7 no.2
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• pp.139-144
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• 2003

Over 350 million people worldwide are chronic carriers of hepatitis B virus (HBV). Chronic viral infections of the liver can progress to cirrhosis, which may ultimately lead to hepatic failure or the development of hepatocellular carcinoma. There are two antiviral drugs on the market approved for clinical management of chronic HBV infections; interferon-alpha and the nucleoside analog lamivudine. However, they showed adverse side-effects. In the rational drug design for such therapies we would like to utilize antiviral drugs that inhibit the HBV replication in the liver. Investigation of natural extracts of silkworm exhibiting antiviral potential was held in the functional HBV polymerase activity and the release of virion particle in the HepG2.2.15 cell lines. HBV-producing transgenic mouse fed with silkworm DNJ molecule was shown as an inhibitor of serum HBV particles. We could represent this DNJ molecule as an antiviral potential complementing conventional therapies after preclinical tests against WHBV-infected animal model, woodchuck.