• Journal of Applied Biological Chemistry
• /
• v.58 no.3
• /
• pp.219-226
• /
• 2015

The anticancer activity of a methanolic extract from lemon leaves (MLL) was assessed in MCF-7-SC human breast cancer stem cells. MLL induced apoptosis in MCF-7-SC, as evidenced by increased apoptotic body formation, sub-G1 cell population, annexin V-positive cells, Bax/Bcl-2 ratio, as well as proteolytic activation of caspase-9 and caspase-3, and degradation of poly (ADP-ribose) polymerase (PARP) protein. Concomitantly, MLL induced the formation of acidic vesicular organelles, increased LC3-II accumulation, and reduced the activation of Akt, mTOR, and p70S6K, suggesting that MLL initiates an autophagic progression in MCF-7-SC via the Akt/mTOR pathway. Epithelial-mesenchymal transition (EMT), a critical step in the acquisition of the metastatic state, is an attractive target for therapeutic interventions directed against tumor metastasis. At low concentrations, MLL induced anti-metastatic effects on MCF-7-SC by inhibiting the EMT process. Exposure to MLL also led to an increase in the epithelial marker E-cadherin, but decreased protein levels of the mesenchymal markers Snail and Slug. Collectively, this study provides evidence that lemon leaves possess cytotoxicity and anti-metastatic properties. Therefore, MLL may prove to be beneficial as a medicinal plant for alternative novel anticancer drugs and nutraceutical products.

• Journal of Applied Biological Chemistry
• /
• v.62 no.4
• /
• pp.375-384
• /
• 2019

The root of Panax ginseng C.A. Meyer were extracted with 70% aqueous EtOH and the concentrates were partitioned into MeOH and H₂O fractions using Diaion HP-20. The repeated SiO₂ or octadecyl SiO₂ column, and MPLC for the MeOH fraction led to isolation of four malonyl ginsenosides. The chemical structures of these compounds were determined as malonyl ginsenoside Rd (1) malonyl ginsenoside Rc (2) malonyl ginsenoside Rb2 (3) malonyl ginsenoside Rb1 (4) based on spectroscopic analyses including Nuclear magnetic resonance and HR-TOF/MS. The contents of malonyl ginsenoside Rb1 was highist as 5.44 mg/g of five years of ginseng. And malonyl ginsenoside Rd was lowest as 0.11 mg/g of six years of ginseng. Additionally, the malonyl ginsenoside Rd exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

• Journal of Applied Biological Chemistry
• /
• v.62 no.4
• /
• pp.399-406
• /
• 2019

The roots of Scrophularia buergeriana were extracted with 80% aqueous Methanol and the concentrates were partitioned into EtOAc, n-BuOH, and H₂O fractions. The repeated silica gel or octadecyl SiO₂column, and medium pressure liquid chromatographies for the n-BuOH fraction led to isolation of phenylethanoid glycosides and iridoid glycosides. The chemical structures of these compounds were determined as harpagoside (1), angoroside C (2), aucubin (3) and acetoside (4) based on spectroscopic analyses including nuclear magnetic resonance and MS. A simple and efficient HPLC with UV detection method for the simultaneous determination of the four compounds (1-4) has been developed and applied to their content determination in the S. buergeriana. The roots were extracted by 80% methanol, and the contents of 1, 2, 3, and 4 were determined to 11.5, 7.6, 41.2, and 4.8 mg/g, respectively. Additionally, angoroside C (2) and acetoside (4) exhibited hepatoprotective effect against ethanol-induced hepatotoxicity in HepG2 cell line.

• Korean Journal of Animal Reproduction
• /
• v.24 no.1
• /
• pp.59-67
• /
• 2000

This study was investigated the developmental potential of bovine embryos following nuclear transfer with bovine fetal fibroblasts (BFF). BFF were isolated from a male 45-day-old-fetus. Non-starved BFF labeled with MitoTracker were transferred into perivitelline space of enucleated oocytes. BFF-oocyte units were fused by electric pulse, and then fused oocytes were activated with calcium ionophore A23187 and subsequently 6-dimethylaminopurine (6-DMAP). The resulting zygotes were placed into CRlaa bovine embryo culture medium. Transfer of the nucleus into enucleated oocyte led to premature chromosome condensation, swelling and pronucleus formation. Remodeled oocytes were developed to the mitotic and 2-cell stage at 18 to 26 h after nuclear transfer. The incidence of in vitro development to the blastocyst stages was 21% of fused oocytes. Mitochondria of BFF eliminated rapidly and were not detected at 8 h after fusion. These results suggest that BFF can be successfully reprogrammed in enucleated bovine oocytes, and that reconstructed embryos can develop to the blastocyst stage.

• Korean Journal of Environmental Biology
• /
• v.32 no.4
• /
• pp.344-352
• /
• 2014

The aim of the present study was to assess the effects of ammonia on physiological responses in Korean rockfish (Sebastes schlegeli). Interestingly, no mortality were observed when the specimens ($301.1{\pm}8.0g$) were exposed to five levels of un-ionized ammonia ($NH_3$) (control, 1, 2, 4, $8mg\;L^{-1}$) for 3 hours. Furthermore, a significantly higher increase in gill $Na^+/K^+$-ATPase (NKA) pump activity with was detected due to the ammonia exposure. The activity of the fishes were found to be 4 and $8mg\;L^{-1}$ $NH_3$, which was significantly high compared to normal $1mg\;L^{-1}$ $NH_3$. Although ammonia exposure had no effect on plasma $Cl^-$, exposure to both 4 and $8mg\;L^{-1}$ $NH_3$, however it led to increase in the plasma $Na^+$, $K^+$ and osmolality levels. Also, prolong ammonia exposure cause increase of plasma cortisol and glucose levels. The increase in glucose was accompanied by an increase in cortisol. The fish exposed to 4 and $8mg\;L^{-1}$ $NH_3$ showed significantly higher hematocrit than control group than those exposed to 1 and $2mg\;L^{-1}$ $NH_3$. The intensity of cell damage increased with the increase concentration and exposure to ammonia. Furthermore, hyperplasia, separation and epithelial necrosis were also observed in gill tissues. Taken together, the results showed that direction of changes to the investigated parameters can be used to determine the physiological responses of Korean rockfish to ammonia.

• Applied Biological Chemistry
• /
• v.34 no.1
• /
• pp.61-66
• /
• 1991

In order to improve the productivity of ethanol by sugar-alcohol-tolerant Saccharomyces cerevisiae D1, the effect of addition of soybean meal on the alcohol fermentation was investigated. The addition of soybean meal led tn the increase of the ethanol productivity and viable cell concentration. Increasing the mont of soybean meal increased the number of viable cells and the consumption percentage of glucose. The water-soluble fraction of soybean meal was nearly as effective as whole-soybean meal, whereas the lipidic fraction had no positive effect. The addition of 4% soybean meal increased the rate of ethanol production regardless of the initial concentrations of glucose. The rate of glucose consumption fermenting a soybean meal supplemented medium was higher than possible in a non-supplemented medium, either in the absence or in the presence of ethanol. But the percentage of ethanol inhibition of the glucose consumption rate was identical for supplemented md unsupplemented media. The increase of final ethanol concentration could not be attributed In an increase of ethanol tolerance of yeast cells but to the satisfaction of nutritional deficiencies.

• Journal of Life Science
• /
• v.21 no.3
• /
• pp.349-357
• /
• 2011

Inhibitors of EGFR (epidermal growth factor receptor) kinase activity may prove useful to therapeutically intervene in cancer and to treat other proliferative diseases. In this study, we investigated the inhibitive effects of two compounds named 63013 and 63033 possess a [1,4]-dioxino quinazoline structure that links the alkoxy side chains together and their structural characteristics are considered to allow better solubility than the dialkoxyquinazoline derivatives. The EGFR kinase activities of A431 human epidermoid carcinoma cells, stimulated by EGF were inhibited by treatment with 63013 and 63033 in a dose-dependent manner respectively. Consistent with the compound-mediated EGFR kinase suppression, the major EGF-related downstream target molecules, such as MEK1/2, MAPK p44/42, AKT and STAT3, were also suppressed by both compounds. Interestingly, both compounds led to cell growth inhibition at a lower concentration than that of Gefitinib (Iressa$^{(R)}$). Collectively, our study showed that both compounds may have good therapeutic potential as an EGFR kinase specific inhibitor to treat EGFR-related diseases.

• Korean Journal of Materials Research
• /
• v.20 no.10
• /
• pp.501-507
• /
• 2010

Changes in surface morphology and roughness of dc sputtered ZnO:Al/Ag back reflectors by varying the deposition temperature and their influence on the performance of flexible silicon thin film solar cells were systematically investigated. By increasing the deposition temperature from $25^{\circ}C$ to $500^{\circ}C$, the grain size of Ag thin films increased from 100 nm to 1000 nm and the grain size distribution became irregular, which resulted in an increment of surface roughness from 6.6 nm to 46.6 nm. Even after the 100 nm thick ZnO:Al film deposition, the surface morphology and roughness of the ZnO:Al/Ag double structured back reflectors were the same as those of the Ag layers, meaning that the ZnO:Al films were deposited conformally on the Ag films without unnecessary changes in the surfacefeatures. The diffused reflectance of the back reflectors improved significantly with the increasing grain size and surface roughness of the Ag films, and in particular, an enhanced diffused reflectance in the long wavelength over 800 nm was observed in the Ag back reflectors deposited at $500^{\circ}C$, which had an irregular grain size distribution of 200-1000 nm and large surface roughness. The improved light scattering properties on the rough ZnO:Al/Ag back reflector surfaces led to an increase of light trapping in the solar cells, and this resulted in a noticeable improvement in the $J_{sc}$ values from 9.94 mA/$cm^2$ for the flat Ag back reflector at $25^{\circ}C$ to 13.36 mA/$cm^2$ for the rough one at $500^{\circ}C$. A conversion efficiency of 7.60% ($V_{oc}$ = 0.93, $J_{sc}$ = 13.36 mA/$cm^2$, FF = 61%) was achieved in the flexible silicon thin film solar cells at this moment.

• Archives of Pharmacal Research
• /
• v.24 no.2
• /
• pp.126-135
• /
• 2001

Quinacrine (QU), a phospholipase-A2 (PLA-2) inhibitor has been used clinically as a chemotherapeutic adjuvant. To understand the mechanisms leading to its chemotherapeutic effect, we have investigated QU-induced apoptotic signaling pathways in human cervical squamous carcinoma HeLa cells. In this study, we found that QU induced cytochrome c-dependent apoptotic signaling. The release of pro-apoptotic cytochrome c was QU concentration- and time-dependent, and preceded activation of caspase-9 and -3. Flow cytometric FACScan analysis using fluorescence intensities of $DiOC_6$/ demonstrated that QU-induced cytochrome c release was independent of mitochondrial permeability transition (MPT), since the concentrations of QU that induced cytochrome c release did not alter mitochondrial membrane potential (${\blacktriangle}{\Psi}_m$). Moreover, kinetic analysis of caspase activities showed that cytochrome c release led to the activation of caspase-9 and downstream death effector caspase-3, Caspase-3 inhibitor (Ac-DEVD-CHO) partially blocked QU-induced apoptosis, suggesting the importance of caspase-3 in this apoptotic signaling mechanism. Supplementation with arachidonic acid (AA) sustained caspase-3 activation induced by QU. Using inhibitors against cellular arachidonate metabolism of lipooxygenase (Nordihydroxyguaiaretic Acid, NDGA) and cyclooxygenase (5,8,11,14-Eicosatetraynoic Acid, ETYA) demonstrated that QU-induced apoptotic signaling may be dependent on its role as a PLA-2 inhibitor. Interestingly, NDCA attenuated QU-induced cytochrome c release, caspase activity as well as apoptotic cell death. The blockade of cytochrome c release by NDCA was much more effective than that attained with cyclosporin A (CsA), a MPT inhibitor. ETYA was not effective in blocking cytochrome c release, except under very high concentrations. Caspase inhibitor z-VAD blocked the release of cytochrome c suggesting that this signaling event is caspase dependent, and caspase-8 activation may be upstream of the mitochondrial events. In summary, we report that QU induced cytochrome c-dependent apoptotic signaling cascade, which may be dependent on its role as a PLA-2 inhibitor. This apoptotic mechanism induced by QU may contribute to its known chemotherapeutic effects.

• Korean Journal of Agricultural Science
• /
• v.13 no.2
• /
• pp.157-167
• /
• 1986

The study was carried out to identify several factors affecting isolation and culture of cotyledone and leaf mesophyll protoplasts of cabbage (Brassica oleraceae), petsai (B.campestris subsp. pekinensis) and rape (B.napus). High viable protoplasts could be obtained when the cotyledon and the leaf mesophyll tissue of all species were treated with enzyme solution composed of 1% macerozyme 'R-10', 1.5% Onozuka 'R-10', 10% mannitol and 50,0 mM $CaCl_22H_2O$ for 4 hours. The protoplasts which obtained from the cotyledon of all species except the cabbage and the leaf mesophyll tissue of all species were divided on NN culture medium supplemented with 9.1% mannitol, 1% glucose, 1% sucrose, $1mg/{\ell}$ 2,4-D, $0.5mg/{\ell}$ NAA and $0.5mg/{\ell}$ BA. The division of the rape leaf mesophyll protoplasts were continued and led to colony.