• The Korean Journal of Pharmacology
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• v.31 no.1 s.57
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• pp.103-114
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• 1995

Platinum coordination complexes are currently one of the most compounds used in the treatment of solid tumors. However, its use is limited by severe side effects such as renal toxicity. Our platinum-based drug discovery program is aimed at developing drugs capable of diminishing toxicity and improving antitumor activity. We synthesized new Pt (Ⅱ) complex analogue containing 1,2-diaminocyclohexane (dach) as carrier ligand and 1,2-bis(diphenylphosphino) ethane (DPPE) as a leaving group. Furthermore, nitrate was added to improve the solubility. A new series of [Pt(trans-ddach)(DPPE).$2NO_3(PC)$ was synthesized and characterized by their elemental analysis and by various spectroscopic techniques [infrared (IR), $^{13}carbon$ nuclear magnetic resonance (NMR)]. PC demonstrated acceptable antitumor activity aganist P388, L-1210 lymphocytic leukemia cells and SK=OV3 human ovarian adenocarcinoma cells, and significant. activity as compared with that. cisplatin. The toxicity of PC was found quite less than thar of cisplatin using MTT, $[^3H]$ thymidine uptake and glucose consumption tests in rabbit proximal tubule cells, human kidney cortical cells and human renal cortical tissues. Based on these results, this novel platinum compound represent a valuable lead in the development of a new anticancer chemotherapeutic agent capable of improving antitumor activity and low toxicity.

• Reproductive and Developmental Biology
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• v.31 no.2
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• pp.83-90
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• 2007

Insulin-like growth factor II (IGF2) and H19 genes are mutually imprinted genes which may be responsible for abnormalities in the cloned fetuses and offspring. This study was performed to identify putative differentially methylated regions (DMRs) of porcine H19 locus and to explore its genomic imprinting in in vitro fertilized (IVF) and somatic cell nuclear transferred (SCNT) embryos. Based on mice genomic data, we identified DMRs on H19 and found porcine H19 DMRs that included three CTCF binding sites. Methylation patterns in IVF and SCNT embryos at the 2-, 4-, $8{\sim}16$-cells and blastocyst stages were analyzed by BS (Bisulfite Sequencing)-PCR. The CpGs in CTCF1 was significantly unmethylated in the 2-cell stage IVF embryos. However, the 4- (29.1%) and $8{\sim}16$-cell (68.2%) and blastocyst (48.2%) stages showed higher methylation levels (p<0.01). On the other hand, SCNT embryos were unmethylayted ($0{\sim}2%$) at all stages of development. The CpGs in CTCF2 showed almost unmethylation levels at the 2-,4- and $8{\sim}16$-cell and blastocyst stages of development in both IVF ($0{\sim}14.1%$) and SCNT ($0{\sim}6.4%$) embryos. At all stages of development, CTCF3 was unmethylated in IVF ($0{\sim}17.3%$) and SCNT ($0{\sim}1.2%$) embryos except at the blastocyst stage (54.5%) of IVF embryos. In conclusion, porcine SCNT embryos showed an aberrant methylation pattern comprised to IVF embryos. Therefore, we suggest that the aberrant methylation pattern of H19 loci may be a reason for increased abnormal fetus after embryo transfer of porcine SCNT embryos.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.86-86
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• 2003

Research has been in progress for more than a decade to production of useful proteins by genetic modification in cattle. However, the levels of protein production in transgenic cattle have been reported very low. To enhance protein production in transgenic animal, we tried homologous recombination to donor cells for production of transgenic clone cattle through nuclear transfer procedure. Thus, we constructed the two targeting vectors of human thrombopoietin (TPO) at bovine $\beta$-casein locus using homologous recombination with 13.6 kb and 9.6 kb homology. In two targeting vectors, positive selection was through the neomycin resistance gene and negative selection was by the diphtheria toxin (DT). Gene targeting was attempted in bovine embryonic fibroblasts (bEF) and bovine ear skin fibroblasts (bESF). To determine the most appropriate concentration of neomycin for bEF and bESF, G4l8 resistance was confirmed by culturing the cells in various concentrations of the drug and both of the cells were optimally selected at $900 \mu g/ml$ of neomycin. The transfected bEF and bESF by the targeting vectors were colonized efficiently at the ratio of DNA to transfection reagent such as $4 \mu g$:2 ${mu}ell$ and $1 \mu g$:$2 \mu l$. Comparing number of healthy clones from passage 4 to passage 8, bESF (17%) persist in culture for much longer than bEF (6%). The two gene-targeted bESF clones of 30 random-integrated clones with 9.6 kb homology length were confirmed, however, nothing was out of 72 random integration clones with 13.6 kb homology length, The DT also worked more efficiently in clones transfected with the vector of 9.6 kb homology length. Our data suggests that the choice of donor cell for long culture period should be considered to obtain targeted cell clone, and the gene-targeting frequency and the DT working efficiency are dependent on the length of target homology.

• Applied Biological Chemistry
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• v.32 no.4
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• pp.393-400
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• 1989

The amounts of $^{14}CO_2$ evolved during the $^{14}C-bentazon$ aging in soil for 3 and 6 months were 6.1 and 14.8% of the original radioactivity, respectively. The presence of earthworms in soil tended to increase the uptake of $^{14}C-bentazon$ by the roots of rice plants, even if it was not statistically significant. The evolution of $^{14}CO_2$ from $^{14}C-bentazon$ in soil increased in the presence of rice plants and earthworms compared with in the absence of them. The uptake of $^{14}C-bentazon$ residues by rice plants decreased remarkably with increasing the aging period within the limit of 3 months both in the absence and presence of earthworms, but there is not much difference between 3-month-aging and 6-month-aging. Much larger amounts of $^{14}C-labelled$ compounds were translocated to the shoots, compared with the data from a previous investigation using maize plants. The amount of non-extractable bound residue increased remarkably with the aging period up to 3 months. The polarity of the compounds extracted from soil increased with the aging and the growing of rice plants, indicating the formation of some polar metabolites.

• Journal of the Korea Institute of Building Construction
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• v.19 no.6
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• pp.477-484
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• 2019

The purpose of this study is to present basic data for developing concrete with EMP shielding as the structure materials when constructing an EMP shielded building structure. In order to use metal-based recycled aggregates with excellent conductivity and easy procurement for EMP shielding concrete, an evaluation of the stability evaluation and EMP shielding performance was performed. Through the stability evaluation, it was found that the coarse aggregate stability criterion was satisfied, but the oxidized slag did not satisfy the fine aggregate stability criterion, the oxidized slag is not satisfied. In addition, as a result of fresh concrete, the workability is increased and the air volume is decreased. The compressive strength is increased due to the high density and coarse granularity of the recycled aggregates, which increased the cement paste and adhesion, thereby increasing the compressive strength. The results of an EMP shielding test show that aggregates with high shielding performance are electronic arc furnace(EAF) Oxidizing Slag and Cooper Slag. The shielding performance is expected to increase if the average particle size of aggregate is small or uniformly distributed.

• 한국생물공학회:학술대회논문집
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• 2003.04a
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• pp.40-44
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• 2003

Soil-borne infectious disease including Pythium aphanidermatum and Rhizoctonia solani causes severe damage to plants, such as cucumber. This soil-borne infectious disease was not controlled effectively by chemical pesticide. Since these diseases spread through the soil, chemical agents are usually ineffective. Instead, biological control, including antagonistic microbe can be used as a preferred control method. An efficient method was developed to select an antagonistic strain to be used as a biological control agent strain. In this new method, surface tension reduction potential of an isolate was included in the ‘decision factor’ in addition to the other factors, such as growth rate, and pathogen inhibition rate. Considering these 3 decision factors by a statistical method, an isolate from soil was selected and was identified as Bacillus sp. GB16. In the pot test, this strain showed the best performance among the isolated strains. The lowest disease incidence rate and fastest seed growth was observed when Bacillus sp. GB16 was used. Therefore this strain was considered as plant growth promoting rhizobacteria (PGPR). The action of surface tension reducing component was deduced as the enhancement of wetting, spreading, and residing of antagonistic strain in the rhizosphere. This result showed that new selection method was significantly effective in selecting the best antagonistic strain for biological control of soil-borne infectious plant pathogen. The antifungal substances against P. aphanidermatum and R. solani were partially purified from the culture filtrates of Bacillus sp. GB16. In this study, lipopeptide possessing antifungal activity was isolated from Bacillus sp. GB16 cultures by various purification procedures and was identified as a surfactin-like lipopeptide based on the Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR), high performance liquid chromatography mass spectroscopy (HPLC-MS), and quadrupole time-of-flight (Q-TOF) ESI-MS/MS data. The lipopeptide, named GB16-BS, completely inhibited the growth of Pythium aphanidermatum, Rhizoctonia solani, Penicillium sp., and Botrytis cineria at concentrations of 10 and 50 mg/L, respectively. A novel method to prevent the foaming and to provide oxygen was developed. During the production of surface active agent, such as lipopeptide (surfactin), large amount of foam was produced by aeration. This resulted in the carryover of cells to the outside of the fermentor, which leads to the significant loss of cells. Instead of using cell-toxic antifoaming agents, low amount of hydrogen peroxide was added. Catalase produced by cells converted hydrogen peroxide into oxygen and water. Also addition of corn oil as an oxygen vector as well as antifoaming agent was attempted. In addition, Ca-stearate, a metal soap, was added to enhance the antifoam activity of com oil. These methods could prevent the foaming significantly and maintained high dissolved oxygen in spite of lower aeration and agitation. Using these methods, high cell density, could be achieved with increased lipopeptide productivity. In conclusion to produce an effective biological control agent for soil-borne infectious disease, following strategies were attempted i) effective screening of antagonist by including surface tension as an important decision factor ii) identification of antifungal compound produced from the isolated strain iii) novel oxygenation by $H_2O_2-catalase$ with vegetable oil for antifungal lipopeptide production.

• Journal of the Korean Society of Food Science and Nutrition
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• v.45 no.4
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• pp.613-618
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• 2016

1,2,3,4,6-Penta-O-galloyl-${\beta}$-D-glucose (PGG) is a gallotannin isolated from various plants such as Galla Rhois. In a previous study, it was reported that PGG has anti-allergic effects by inhibiting interleukin (IL)-4 signaling in B cells. However, the effect of PGG on basophilic cells remains unclear. Therefore, the aim of this study was to investigate the inhibitory effect of PGG on mitogen and calcium ionophore-induced allergic responses. PGG had no effect on proliferation and cytotoxicity of RBL-2H3 cells. PGG significantly suppressed cell degranulation (histamine and ${\beta}-hexosaminidase$) as well as inflammatory cytokine production such as IL-4 and tumor necrosis factor-${\alpha}$. The underlying mechanism of PGG on these anti-allergic actions was correlated with inhibition on translocation of nuclear factor-${\kappa}B$ from the cytosol to nucleus. These data suggest that PGG is a potentially effective functional compound for prevention of allergic diseases.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.10a
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• pp.37-43
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• 2001

1. About fifty thousand of cattle embryos were transferred and 16000 ET-calves were born in 1999. Eighty percents of embryos were collected from Japanese Black beef donors and transferred to dairy Holstein heifers and cows. Since 1985, we have achieved in bovine in vitro fertilization using immature oocytes collected from ovaries of slaughterhouse. Now over 8000 embryos fertilized by Japanese Black bull, as Kitaguni 7~8 or Mitsufuku, famousbulls as high marbling score of progeny tests were sold to dairy farmers and transferred to their dairy cattle every year. 2. Embryo splitting for identical twins is demonstrated an useful tool to supply a bull for semen collection and a steer for beef performance test. According to the data of Dr. Hashiyada(2001), 296 pairs of split-half embryos were transferred to recipients and 98 gave births of 112 calves (23 pairs of identical twins and 66 singletons). 3. A blastomere-nuclear-transferred cloned calf was born in 1990 by a joint research with Drs. Tsunoda, National Institute of Animal Industry (NIAI) and Ushijima, Chiba Prefectural Farm Animal Center. The fruits of this technology were applied to the production of a calf from a cell of long-term-cultured inner cell mass (1988, Itoh et al, ZEN-NOH Central Research Institute for Feed and Livestock) and a cloned calf from three-successive-cloning (1997, Tsunoda et al.). According to the survey of MAFF of Japan, over 500 calves were born until this year and a glaf of them were already brought to the market for beef. 4. After the report of "Dolly", in February 1997, the first somatic cell clone female calves were born in July 1998 as the fruits of the joint research organized by Dr. Tsunoda in Kinki University (Kato et al, 2000). The male calves were born in August and September 1998 by the collaboration with NIAI and Kagoshima Prefecture. Then 244 calves, four pigs and a kid of goat were now born in 36 institutes of Japan. 5. Somatic cell cloning in farm animal production will bring us as effective reproductive method of elite-dairy- cows, super-cows and excellent bulls. The effect of making copy farm animal is also related to the reservation of genetic resources and re-creation of a male bull from a castrated steer of excellent marbling beef. Cloning of genetically modified animals is most promising to making pig organs transplant to people and providing protein drugs in milk of pig, goat and cattle. 6. Farm animal cloning is one of the most dreamful technologies of 21th century. It is necessary to develop this technology more efficient and stable as realistic technology of the farm animal production. We are making researches related to the best condition of donor cells for high productivity of cloning, genetic analysis of cloned animals, growth and performance abilities of clone cattle and pathological and genetical analysis of high rates of abortion and stillbirth of clone calves (about 30% of periparutum mortality). 7. It is requested in the report of Ministry of Health, labor and Welfare to make clear that carbon-copy cattle(somatic cell clone cattle) are safe and heathy for a commercial market since the somatic cell cloning is a completely new technology. Fattened beef steers (well-proved normal growth) and milking cows(shown a good fertility) are now provided for the assessment of food safety.

• Journal of Life Science
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• v.28 no.6
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• pp.733-737
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• 2018

Cucurbitacin-I, a natural triterpenoid derived from Cucurbitaceae family plants, exhibits a number of potentially useful pharmacological and biological activities. Indeed, the previous study demonstrated that cucurbitacin-I reduced the proliferation of colon cancer cells by enhancing apoptosis and causing cell cycle arrest at the G2/M phase. CD44, a type I transmembrane protein with the function of adhering to cells, mediates between the extracellular matrix and other cells through hyaluronic acid. Recent studies have demonstrated that an overexpression of the CD44 membrane receptor results in tumor initiation and growth, specific behaviors of cancer stem cells, the development of drug resistance, and metastasis. The aim was to examine the effect of cucurbitacin-I on CD44 expression human ovarian cancer cells because the effect of cucurbitacin-I on CD44 expression has not been reported. The expressions of CD44 mRNA and protein were detected using a quantitative real-time reverse-transcription polymerase chain reaction and a Western blot analysis, respectively. Treatment with cucurbitacin-I inhibited the expression of CD44 mRNA and protein. A subsequent analysis revealed that cucurbitacin-I blocked the phosphorylation of activator protein-1 (AP-1) and nuclear factor kappa-B ($NF-{\kappa}B$), which are key regulators of CD44 expression. Taken together, the data demonstrate that cucurbitacin-I regulates the AP-1 and $NF-{\kappa}B$ signaling pathways, leading to decreased CD44 expression.

• Journal of Embryo Transfer
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• v.22 no.3
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• pp.179-184
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• 2007

Culture method of somatic cells is one of the important factors in the production of transgenic pigs by somatic cell nuclear transfer. In this study, we established an efficient culture method of porcine fetal fibroblasts. Porcine fetal fibroblasts were isolated from 33-day-old fetuses. The proliferation of porcine fetal fibroblasts was analyzed by different serum types and culture media. The cultures in medium supplied 15% ES screened FBS showed faster increase in cell number than 15% FBS. Also, fetal fibroblasts have been propagated continuously for $7{\sim}8$ passages in ES modified DMEM and DMEM medium. We transfected $PGK-neo^r$ vector (pKJ2) into porcine fetal fibroblasts to estimate colony formation in this culture condition. The formation of colonies was confirmed in the medium containing $300\;{\mu}g/ml$ G418 at 12 day. These data show that this culture system can be used screening of porcine somatic cells transfected transgene.