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Biological activity of Euonymus alatus (Thunb.) Sieb. wing extracts (화살나무 날개 추출물의 생리활성)

  • Hye-Ji Min;Du-Hyun Kim;Kwon-Il Seo
    • Food Science and Preservation
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    • v.30 no.2
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    • pp.358-368
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    • 2023
  • Euonymus alatus (Thunb.) Sieb., also known as the arrow tree in Korea, is a plant in East Asia used in traditional medicine and food. In particular, the wings of E. alatus are rich in phenolic compounds. This study evaluated the antioxidant, α-glucosidase inhibition, and anti-cancer activities of E. alatus wing extracts. The radical and hydrogen peroxide scavenging acitvities and reducing the power of 1,000 ㎍/mL E. alatus wing extracts, were similar to those of the positive control (0.1% BHT, 0.1% α-tocopherol). In addition, ethanol and methanol extract at 250 ㎍/mL showed 95.70 and 94.99% of α-glucosidase inhibition activity, respectively. The ethanol extract of E. alatus wings had the highest total polyphenol and flavonoid contents (867.8 mg% and 521.7 mg%, respectively). The E. alatus wing extracts significantly decreased the cell viability of LNCaP human prostate cancer cells (p<0.001), MDA-MB-231 human breast cancer cells (p<0.001), and HT-29 human colon cancer cells (p<0.001) in a dose-dependent manner. However, there was no significant effect on B16 mouse melanoma cells. Notably, the ethanol extracts showed higher cancer cell growth inhibitory activity in LNCaP and HT-29 cells than the other extracts. These results suggest that E. alatus wing extracts could have significant clinical applications, and our results can be used as basic data for future functional food material development.

Somatic Cells Count and Its Genetic Association with Milk Yield in Dairy Cattle Raised under Thai Tropical Environmental Conditions

  • Jattawa, D.;Koonawootrittriron, S.;Elzo, M.A.;Suwanasopee, T.
    • Asian-Australasian Journal of Animal Sciences
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    • v.25 no.9
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    • pp.1216-1222
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    • 2012
  • Somatic cells count (SCC), milk yield (MY) and pedigree information of 2,791 first lactation cows that calved between 1990 and 2010 on 259 Thai farms were used to estimate genetic parameters and trends for SCC and its genetic association with MY. The SCC were log-transformed (lnSCC) to make them normally distributed. An average information-restricted maximum likelihood procedure was used to estimate variance components. A bivariate animal model that considered herd-yr-season, calving age, and regression additive genetic group as fixed effects, and animal and residual as random effects was used for genetic evaluation. Heritability estimates were 0.12 (SE = 0.19) for lnSCC, and 0.31 (SE = 0.06) for MY. The genetic correlation estimate between lnSCC and MY was 0.26 (SE = 0.59). Mean yearly estimated breeding values during the last 20 years increased for SCC (49.02 cells/ml/yr, SE = 26.81 cells/ml/yr; p = 0.08), but not for MY (0.37 kg/yr, SE = 0.87 kg/yr; p = 0.68). Sire average breeding values for SCC and MY were higher than those of cows and dams (p<0.01). Heritability estimates for lnSCC and MY and their low but positive genetic correlation suggested that selection for low SCC may be feasible in this population as it is in other populations of dairy cows. Thus, selection for high MY and low SCC should be encouraged in Thai dairy improvement programs to increase profitability by improving both cow health and milk yield.

Role of cAMP, EGF, IGF-I and Protein Phosphorylation in Mammary Development I. Effect of EGF, IGF-I and Photoreactive Cyclic AMP on DNA Synthesis of Mammary Epithelial Cell (유선발달에 있어서 cAMP, EGF, IGF-I 및 단백질 인산화 작용의 역할 I. EGF, IGF-I 및 Photoreactive Cyclic AMP가 유선상피세포의 DNA합성에 미치는 효과)

  • 여인서;박춘근;홍병주
    • Korean Journal of Animal Reproduction
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    • v.17 no.1
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    • pp.49-56
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    • 1993
  • Mouse mammary epithelial cells(NMuMG) were plated onto 24 well phates(100,000 cells/well), in DMEM supplemented with 10% fetal calf serum. After serum starvation for 24 hours, EGF)0~100ng/ml) was added simultaneously with IGF-I(10ng/ml), 1$\mu$M photoreactive cAMP(4,5-dimethoxy-2-nitrobenzyl adenosine-3',5' cyclic monophosphate, DMNB) or IGF-I plus DMNB. After 2 hours, the cells were expposed to UV light(300nm, 3 second pulse0 in order to activate DMNB which induces a rapid transient increase in intracellular cAMP upon UV irradiation. DNA synthesis was estimated as incorporation of 3H-thymidine into DNA(1 hour pulse with 1$\mu$Ci/ml, 18~19 hours after UV exposure). Without IGF-I or DMNB, EGF(10 or 100ng/ml) increased DNA synthesis from 8,362 dpm/well in control to 16,345 or 18,684 dpm/well with EGF(pooled SE=1,239 dpm/well, P<0.05). IGF-I or IGF-I plus DMNB alone increased DNA synthesis from 8,362 dpm/well in control to 17,307 or 20,427 dpm/well, respectively(P<0.05). Addition of IGF-I, DMNB or IGF-I plus DMNB into 0~100ng/ml EGF did not significantly change the shape of dose response curve of EGF alone. In other experiment, EGF or IGF-I plus DMNB into 10ng/ml EGF group exhibited interaction effect in DNAsynthesis [EGF(10ng/ml)=18,497; IGF-I+EGF=22,837; DMNB+EGF=20,658 ; IGF-I+DMNB+EGF=29,658, pooled SE=1,055, P<0.05]. These results indicate that simultaneous activation of EGF, IGF-I and intracellular cAMP interact in DNA synthesis of mouse mammary epithelial cells.

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Studies on the Optimal Culture Conditions for Itaconic Acid Production by Aspergilus Terreus (Aspergillus Terreus에 의한 이타콘산 생산을 위한 최적배양조건에 관한 연구)

  • 박승원;김승욱
    • KSBB Journal
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    • v.9 no.4
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    • pp.436-442
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    • 1994
  • The production of itaconic acid by Aspergillus terreus NRRL 1960 was studied. The optimal culture conditional such as pH, inoculum size and medium composition were established. Maximum production of itaconic acid, $19.18g/\ell$, was obtained when the cultivation was carried out at $37^{\circ}C$ and pH 2.5 for 7days, with medium containing 5%(w/v) glucose, 0.5%(w/v) NH4Cl, 0.2%(w/v) yeast extract 0.1%(w/v) CaC12, 0.1%(w/v) MgSO4 and 0.2%(w/v) NaCl. A proper medium for inoculum culture was found to be 2%(w/v) malt extract. The batch production of itaconic acid with free cells in a stirredtank reactor was not efficient compared to the shake-flask culture.

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A Natural L-Arginine Analog, L-Canavanine-Induced Apoptosis is Suppressed by Protein Tyrosine Kinase p56lck in Human Acute Leukemia Jurkat T Cells (인체 급성백혈병 Jurkat T 세포에 있어서 L-canavanine에 의해 유도되는 세포자살기전에 미치는 단백질 티로신 키나아제 p56lck의 저해 효과)

  • Park, Hae-Sun;Jun, Do-Youn;Woo, Hyun-Ju;Rue, Seok-Woo;Kim, Sang-Kook;Kim, Kyung-Min;Park, Wan;Moon, Byung-Jo;Kim, Young-Ho
    • Journal of Life Science
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    • v.19 no.11
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    • pp.1529-1537
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    • 2009
  • To elucidate further the antitumor effects of a natural L-arginine analogue, L-canavanine, the mechanism underlying apoptogenic activity of L-canavanine and its modulation by protein tyrosine kinase $p56^{lck}$ was investigated in human Jurkat T cells. When the cells were treated with 1.25 to 2.5 mM L-canavanine for 36 h, several apoptotic events including mitochondrial membrane potential (${\Delta\Psi}m$) loss, activation of caspase-9, -3, -8, and -7, poly (ADP-ribose) polymerase (PARP) degradation, and DNA fragmentation were induced without alteration in the levels of Fas or FasL. These apoptotic changes were more significant in $p56^{lck}$-deficient Jurkat clone JCaM1.6 than in $p56^{lck}$-positive Jurkat clone E6.1. The L-canavanine-induced apoptosis observed in $p56^{lck}$-deficient JCaM1.6 cells was significantly reduced by introducing $p56^{lck}$ gene into JCaM1.6 cells by stable transfection. Treatment of JCaM1.6/lck cells with L-canavanine caused a transient 1.6-fold increase in the kinase activity of $p56^{lck}$. Both FADD-positive wild-type Jurkat T cell clone A3 and FADD-deficient Jurkat T cell clone I2.1 exhibited a similar susceptibility to the cytotoxicity of L-canavanine, excluding involvement of Fas/FasL system in triggering L-canavanine-induced apoptosis. The L-canavanine-induced apoptotic sub-$G_1$ peak and activation of caspase-3, -8, and -7 were abrogated by pan-caspase inhibitor (z-VAD-fmk), whereas L-canavanine-induced activation of caspase-9 was not affected. These results demonstrated that L-canavanine caused apoptosis of Jurkat T cells via the loss of ${\Delta\Psi}m$, and the activation of caspase-9, -3, -8, and -7, leading to PARP degradation, and that the $p56^{lck}$ kinase attenuated the ${\Delta\Psi}m$ loss and activation of caspases, and thus contributed as a negative regulator to L-canavanine-induced apoptosis.

Production and Characterization of Monoclonal Antibodies to Yeast Mitochondrial RNA Polymerase Specificity Factor

  • Lee, Chang-Hwan;Jang, Sei-Heon
    • BMB Reports
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    • v.31 no.6
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    • pp.607-610
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    • 1998
  • Transcription of mitochondrial DNA in the yeast S. cerevisiae depends on recognition of a consensus nonanucleotide promoter sequence by mitochondrial RNA polymerase specificity factor, which is a 43 kDa polypeptide encoded by the nuclear MTF1 gene. Mtf1p has only limited amino acid sequence homology to bacterial sigma factors, but functions in many ways like sigma in that it is required for promoter recognition and initiation of transcription. To analyze the corebinding region of Mtf1p, monoclonal antibodies to this protein were prepared. Recombinant Mtf1p overproduced in E. coli was purified to near homogeneity and used to raise monoclonal antibodies (mAbs). From fused cells screened for Mtf1p mAbs by immunodot blot analysis, 19 positive clones were initially isolated. Further analysis of positive clones by Western blotting resulted in 4 mAbs of Mtf1p.

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Effects of the Essential Oil of Nelumbo nucifera Flower on Glioma Cells (련화향(蓮花香) 정유액이 glioma cell에 미치는 효과)

  • Kim, In-Ja;Lee, Joo-Yeon;Choi, Bang-Seob;Kim, Geun-Woo;Koo, Byung-Soo
    • Journal of Oriental Neuropsychiatry
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    • v.19 no.2
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    • pp.111-122
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    • 2008
  • Objective : Herb medicines are potential sources of useful edible and medicinal plants. They are used as a drug because of their various biological activities such as immunomodulatory, antiviral, and antitumor functions. Nelumbo nucifera have been applied in Chinese herbal prescriptions to improve tissue inflammation. However, it has not been elucidated on the effect of the flower of Nelumbo nucifera in cells. Method : In the present study, to examine the effect of that on glioma cells, U87, the essential oil was extracted from the flower of Nelumbo nucifera (NN essential oil). U87 cells were exposed to different concentrations of 2-40 ug/ml of NN essential oil in ethanol. Cell viability was measured by MTT assay at 24 h. To find out the intracellular target signal molecule(s) for this antiproliferative activity of NN essential oil, phosphorylation of Akt, ERM, MAPK or p38 proteins were examined by Western blot analysis. To study long term effect of NN essential oil in U87 cells, the image of cells treated with NN essential oil for 4 days were obtained. Results and Conclusion : NN essential oil was shown to exhibit antitumor activity in glioma cells, at a broad range of concentrations of 10-40 ug/ml. The phosphorylation of Akt and Endoplasmic Reticulum Matrix (ERM) proteins which known to be involved in the cell death, were gradually decreased to 2 hours after addition 20 ug/ml of NN essential oil. However, the phosphorylation of mitogen-activated protein (MAPK) and p38 was found to increase in NN essential oil treated cells. NN essential oil treated cells showed decreased glioma cell number. These results provide a possible NN essential oil-induced inhibitory signal for tumor cell proliferation that is initiated by the decrease in Akt activity. Moreover, it is likely that the activation of p38 is required for the NN essential oil-induced inhibition of tumor proliferation.

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Studies on the Immobilization of Saccharomyces cerevisiae for Ethanol Production (효모의 Alginate 고정화에 관한 연구)

  • 한면수;하상도;정동효
    • Microbiology and Biotechnology Letters
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    • v.19 no.4
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    • pp.390-397
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    • 1991
  • Ethanol production by calcium alginate-immobilized baker's yeast (Saccharor/tyces cereviszae) was studied in the batch fermentation using glucose medium as a feed. Immobilied cells were stable between $30^{\circ}C$ and $40^{\circ}C$ whereas free cells were stable between $30^{\circ}C$ and $37^{\circ}C$ The beads were showed constant ethanol productivity during 720 hours (30 days) over. Fermentation characteristics of immobilized baker's yeast were examined changing the initial glucose concentration of broth in fermentation. Initial glucose concentrations employed were 50, 100, 150 and 200 g/l, respectively. In 15% gucose medium, maximum specific growth rate, maximum ethanol yield and ethanol concentration were observed as 0.092 $h^{-1}$, 0.45, 67.5 g/l, respectively.

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Cloning and Prokaryotic Expression of C-type Lysozyme Gene from Agrius convolvuli

  • Kim, Jong-Wan;Yoe, Sung-Moon
    • Animal cells and systems
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    • v.12 no.3
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    • pp.149-155
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    • 2008
  • We have isolated and characterized Agrius convolvuli cDNA encoding a c-type lysozyme. The cDNA sequence encodes a processed protein of 139 amino acid residues with 19 amino acid residues amino-terminal signal sequence and 120 amino acid residues mature sequence. The amino acid residues responsible for the catalytic activity and the binding of the substrate are conserved. Agrius lysozyme has a high identity to Manduca sexta. Recombinant A. convolvuli lysozyme was expressed in Escherichia coli BL21(DE3) pLysS cells for pGEX 4T-1 expression vector. Their optimal conditions for the fusion protein expression and purification were screened. Lysozyme gene amplified with primers ACLyz BamHI and ACLyz XhoI was ligated into the pGEX 4T-1 vector, which contained the glutathione S-transferase(GST) gene for fusion partner. The fusion protein was induced by IPTG and identified by SDS-PAGE analysis. Molecular weight of the fusion protein was estimated to be about 45 kDa. Recombinant lysozyme, fused to GST, was purified by glutathion-Sepharose 4B affinity chromatography. Western blot analysis of this protein revealed an immunoreactivity with the anti-Agrius lysozyme.

Stimulatory Effect of Saponin from Panax ginseng on Function of Lymnphocytes in the Elderly

  • Liu, Jun-Da;Wang, Shu;Liu, Hong-Tao
    • Proceedings of the Ginseng society Conference
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    • 1998.06a
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    • pp.312-321
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    • 1998
  • We used the saponin Rgi extracted from Panax ginseng to study its effects on lymphocytes of 10 young and 19 elderly persons. The proliferate response of Iymphocytes cocultured for 72h with PHA and saponin was measured by using MTT method and the'H-TdR incorporation procedure. PHA and Rgl had stimulative effects on the phenotype of Iymphocytes (p<0.001). Rgl also increased the fluidity of lymphocyte membrane of the aged (p<0.001). The CD2s and CDfsRA positive cells of Iymphocytes in the elderly were lower than those of the young people,8.6clo $\pm$ 2.7olo vs 10.43% : 3.5%, 20.95% $\pm$ 15.5clQ vs 50.86% :4.3olo, respectively. More CDfsRO positive cell lymphocy populations were seen in the aged. The CEfsRO positive cells of the young people were 39.63% $\pm$ 3.2%. We discussed the cause of declined immune function of Lymphocytes of aged person and the mechanism of the effect of P. ginseng on Lymphocytes. Key words: Saponin, Lymphocytes, Aged person, Stimulatory effect, and Panax ginseng

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