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Detection of Ref-1 (Redox factor-1) Interacting Protein Using the Yeast Two-hybrid System (Yeast two-hybrid system을 이용한 Ref-1 (redox factor-1) 결합 단백질의 분리 및 동정)

  • 이수복;김규원;배문경;배명호;정주원;안미영;김영진
    • Journal of Life Science
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    • v.14 no.1
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    • pp.26-31
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    • 2004
  • Redox factor-1 (Ref-1), known as a redox regulator, controls the DNA binding of AP-1 and is activated in HT29 colon cancer cells by hypoxia in vitro. REF-1 also increases tile DNA binding affinity of Hypoxia-inducible Factor-lalpha$ (HIF-lalpha$), HIF-like Factor (HLF) and early growth response-1 (Egr-1) which induce expression of the genes involved in angiogenesis, so that we speculate that REF-1 may play a role in hypoxia-induced angiogenesis. In this research we tried to detect novel proteins interacting with REF-1 using Yeast two-hybrid system using full-length REF-1 cDNA as bait. As result of such screening we detected 3 positive clones. DNA sequencing and GeneBank search revealed that one of the clones contained the same sequences as M.musculus cDNA for tioredoxin.

Physiological Characteristics and Anti-obesity Effect of Lactobacillus plantarum K10

  • Kim, Seulki;Huang, Eunchong;Park, Soyoung;Holzapfel, Wilhelm;Lim, Sang-Dong
    • Food Science of Animal Resources
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    • v.38 no.3
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    • pp.554-569
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    • 2018
  • This study aimed to investigate the physiological characteristics and anti-obesity effects of Lactobacillus plantarum K10. The ${\alpha}-amylase$ inhibitory activity, ${\alpha}-glucosidase$ inhibitory activity, and lipase inhibitory activity of L. plantarum K10 was $94.66{\pm}4.34%$, $99.78{\pm}0.12%$, and $87.40{\pm}1.41%$, respectively. Moreover, the strain inhibited the adipocyte differentiation of 3T3-L1 cells ($32.61{\pm}8.32%$) at a concentration of $100{\mu}g/mL$. In order to determine its potential for use as a probiotic, we investigated the physiological characteristics of L. plantarum K10. L. plantarum K10 was resistant to gentamycin, kanamycin, streptomycin, ampicillin, ciprofloxacin, tetracycline, vancomycin, and chloramphenicol. It also showed higher Leucine arylamidase, Valine arylamidase, and ${\beta}-galactosidase$ activities. Moreover, it was comparatively tolerant to bile juice and acid, exhibiting resistance to Escherichia coli, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus with rates of 90.71%, 11.86%, 14.19%, and 23.08%, respectively. The strain did not produce biogenic amines and showed higher adhesion to HT-29 cells compared to L. rhamnosus GG. As a result of the animal study, L. plantarum K10 showed significantly lower body weight compared to the high-fat diet group. The administration of L. plantarum K10 resulted in a reduction of subcutaneous fat mass and mesenteric fat mass compared to the high-fat diet (HFD) group. L. plantarum K10 also showed improvement in gut permeability compared to the HFD positive control group. These results demonstrate that L. plantarum K10 has potential as a probiotic with anti-obesity effects.

Anticancer Activity of the Safflower Seeds (Carthamus tinctorius L.) through Inducing Cyclin D1 Proteasomal Degradation in Human Colorectal Cancer Cells

  • Park, Gwang Hun;Hong, Se Chul;Jeong, Jin Boo
    • Korean Journal of Plant Resources
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    • v.29 no.3
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    • pp.297-304
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    • 2016
  • The seed of safflower (Carthamus tinctorius L) has been reported to suppress human cancer cell proliferation. However, the mechanisms by which safflower seed inhibits cancer cell proliferation have remained nuclear. In this study, the inhibitory effect of the safflower seed (SS) on the proliferation of human colorectal cancer cells and the potential mechanism of action were examined. SS inhibited markedly the proliferation of human colorectal cancer cells (HCT116, SW480, LoVo and HT-29). In addition, SS suppressed the proliferation of human breast cancer cells (MDA-MB-231 and MCF-7). SS treatment decreased cyclin D1 protein level in human colorectal cancer cells and breast cancer cells. But, SS-mediated downregulated mRNA level of cyclin D1 was not observed. Inhibition of proteasomal degradation by MG132 attenuated cyclin D1 downregulation by SS and the half-life of cyclin D1 was decreased in SS-treated cells. In addition, SS increased cyclin D1 phosphorylation at threonine-286 and a point mutation of threonine-286 to alanine attenuated SS-mediated cyclin D1 degradation. Inhibition of ERK1/2 by PD98059 suppressed cyclin D1 phosphorylation and downregulation of cyclin D1 by SS. In conclusion, SS has anti-proliferative activity by inducing cyclin D1 proteasomal degradation through ERK1/2-dependent threonine-286 phosphorylation of cyclin D1. These findings suggest that possibly its extract could be used for treating colorectal cancer.

Study on Antitumor Activity of Hwalheuldaibotang(HDBT) (활혈대보탕(活血大補湯)의 항암활성(抗癌活性) 및 항전이(抗轉移) 효과(效果)에 관(關)한 연구(硏究))

  • Bae, Moon-yong;Kim, Dong-hee
    • Journal of Haehwa Medicine
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    • v.9 no.2
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    • pp.97-109
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    • 2001
  • To evaluate the antitumor activity and antimetastatic effects of HDBT, studies were done experimentally. The results were obtained as follows: 1. HDBT extracts didn't show cytotoxicity against BALB/C mouse lung fibroblast cell. 2. In cytotoxicity against A549, SK-OV-3, B16-BL6 and HT1080 concen- tration inhibiting cell growth up to below 30% of control was recognized at $10^{-3}g/ml$ of HDBT. 3. The concentration inhibiting adhesion of A549 and B16-BL6 to complex extracellular matrix up to below 30% of control was recognized at $10^{-3}g/ml$ of HDBT. 4. In Inhibitory effect on activity of DNA topoisomerase I, the $IC_{50}$ was shown $200-300{\mu}g/m{\ell}$ of HDBT. 5. The T/C% was 137.9% in HDBT-treated group in S-180 bearing ICR mice. 6. In CAM assay, HDBT extracts inhibited angiogenesis significantly at $15{\mu}g/egg$ concentration as compared with control. 7. In pumonary colonization assay, a number of colonies in the lungs were decreased but insignificantly in HDBT-treated group as compared with control group. 8. In hematological changes in B16-BL6 injected C57BL/6, numbers of WBC were decreased significantly in HDBT-treated group but numbers PLT were increased insignificantly as compared with control. From above results it was concluded that HDBT could be usefully applied for the prevention and treatment of cancer.

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Cytotoxic and ACAT-inhibitory Sesquiterpene Lactones from the Root of Ixeris dentata forma albiflora

  • Ahn, Eun-Mi;Bang, Myun-Ho;Song, Myoung-Chong;Park, Mi-Hyun;Kim, Hwa-Young;Kwon, Byoung-Mog;Baek, Nam-In
    • Archives of Pharmacal Research
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    • v.29 no.11
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    • pp.937-941
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    • 2006
  • Ixeris dentata forma albiflora was extracted with 80% aqueous MeOH, and the concentrated extract was partitioned with EtOAc, n-BuOH and $H_{2}O$. Eight sesquiterpenes were isolated through repeated silica gel and octadecyl silica gel ($C_{18},\;ODS$) column chromatography of the EtOAc and n-BuOH fractions. Physicochemical analysis using NMR, MS and IR revealed the chemical structures of the sesquiterpenes, which were zaluzanin (1), 9a-hydroxyguaian-4(15), 10(14), 11 (13)-triene-6, 12-olide(2), $3{\beta}-O-{\beta}-D-glucopyranosyl-8{\beta}-hydroxyguaian$-4(15), 10(14)-diene-6, 12-olide (3), $3-O-{\beta}-D-glucopyranosyl-8{\beta}-hydroxyguauan$-10(14)-ene-6, 12-olide (4), ixerin M (5), glucozaluzanin C (6), crepiside I (7), and ixerin D (8). This is the first time that these sesquiterpene lactones have been isolated from this plant. Compounds 1, 2 and 7 revealed relatively high cytotoxicities on human colon carcinoma cell and lung adeno-carcinoma cell, while compounds 5 and 7 showed acyl-CoA: cholesterol acyltransferase (ACAT) inhibitory activity.

Heterotopic electroacupuncture modulates formalin-induced pain via descending inhibition in the rat (백서(白鼠)의 formalin 유발(誘發) 통증(痛症)에 대한 전침자극(電鍼刺戟)과 하행성 진통기전)

  • Koo, Sung-Tae;Sohn, In-Chul;Kim, Jae-Hyo
    • Korean Journal of Acupuncture
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    • v.23 no.3
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    • pp.55-71
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    • 2006
  • Objectives : The present study was investigated the effect and pathway of heterotopic electroacupuncture (EA) on pain induced by formalin in rats. Methods : Acupoints in the right forepaws, $HT_7$ and $PC_7$, were stimulated with 3 mA, 2 ms, and 10 Hz before subcutaneously formalin injection (5%, $50{\mu}l$) to the left hind paw. Moreover, it was investigated whether the dorsolateral funiculus (DLF), as known to the descending inhibition, mediates analgesia of the heterotopic EA, and an administration of naltrexone blocks the effect of EA. Results : In the immunohistochemistry of cFos-like protein (cFL), there were inhibitory effects of EA on the increased expression of cFL in the lumbar spinal dorsal horn neurons following formalin injection. Especially, EA inhibited the expression of cFL on the superficial laminae than that on the deep laminae at 1 hr after, but that on the deep laminae than that on the superficial laminae at 2 hr after. Also, EA suppressed the increased expression of nitric oxide (NO) and neuronal nitric oxide synthase (nNOS) in the lumbosacral spinal cord after formalin injection, but not Sham-EA. Suppressed expressions of cFL, NO and nNOS in the spinal cord were eliminated after transection of the ipsilateral DLF at $T_{10}{\sim}T_{11}$ levels. However, pretreatment of naltrexone could not prevent the suppressive expressions of cFL, NO and nNOS at the spinal cord. Conclusions : These results suggest that the analgesia of heterotopic EA may be modulated through the DLF constituting the descending inhibition.

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Influence of Hot-Extrusion on Mechanical Properties of AZ31B Magnesium Alloy Sheet (AZ31B 마그네슘 합금의 기계적 특성에 미치는 열간압출의 영향)

  • Kim Yong-Gil;Choi Hak-Kyu;Kang Min-Cheol;Jeong Hae-Yong;Bae Cha-Hurn
    • Korean Journal of Materials Research
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    • v.15 no.1
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    • pp.25-30
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    • 2005
  • The microstructural changes by hot extrusion of AZ31B magnesium alloy were observed, and the relation to the tensile property was examined. The tensile properties as oriented longitudinal(L), half transverse(HT) and long transverse(LT) to the extrusion direction were investigated at $20^{\circ}C,\;100^{\circ}C,\;200^{\circ}C,\;300^{\circ}C\;and\;400^{\circ}C$, respectively. As the results, many recrystallized small grains distributed uniformly in large banded microstructures formed along the extrusion direction. The grain size of as-extruded specimen was around $30\~150\;{\mu}m$. As increasing the test temperature the tensile and yield strength with respect to the angle between the axis of the tensile and the longitudinal direction in extrusion was decreased, but their elongation were increased and their deviation between L and LT specimens have disappeared from $300^{\circ}C$. This mechanical anisotropy was reduced at elevated temperatures and almost disappeared at $400^{\circ}C$. It was considered that the homogenization was occured by the recrystallization and the change of slip system was occurred during tensile test process in elevated temperatures.

Microencapsulation of Probiotic Lactobacillus acidophilus KBL409 by Extrusion Technology to Enhance Survival under Simulated Intestinal and Freeze-Drying Conditions

  • Lee, YunJung;Ji, Yu Ra;Lee, Sumi;Choi, Mi-Jung;Cho, Youngjae
    • Journal of Microbiology and Biotechnology
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    • v.29 no.5
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    • pp.721-730
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    • 2019
  • The probiotic Lactobacillus acidophilus KBL409 was encapsulated with alginate (Al) and alginate-chitosan (Al/Chi) through extrusion method. The sizes and zeta potentials of microspheres were measured to confirm encapsulation. To evaluate the protective effect of microspheres against gastrointestinal fluids, all the samples were exposed to simulated gastric fluids (SGFs, pH 1.5) at $37^{\circ}C$ for 1 or 2 h, followed by incubation with simulated intestinal fluids (SIFs, pH 6.5) for 2 h. The mucoadhesive ability of microspheres was evaluated using the intestinal epithelial cell line HT29-MTX. To extend the shelf-life of probiotics, lyoprotectants such as disaccharide and polysaccharide were mixed with free or encapsulated cells during the freeze-drying process. The size of the microspheres demonstrated a narrow distribution, while the zeta potentials of Al and Al/Chi-microspheres were $-17.9{\pm}2.3$ and $20.4{\pm}2.6mV$, respectively. Among all the samples, Al/Chi-encapsulated cells showed the highest survival rate even after exposure to SGF and SIF. The mucoadhesive abilities of Al and Al/Chi-microspheres were higher than 94%, whereas the free L. acidophilus showed 88.1% mucoadhesion. Ten percent of sucrose showed over 80% survival rate in free or encapsulated cells. Therefore, L. acidophilus encapsulated with Al and Al/Chi-microspheres showed higher survival rates after exposure to the gastrointestinal tract and better mucoadhesive abilities than the free cells. Also, sucrose showed the highest protective effect of L. acidophilus during the freeze-drying process.

Cr Electroplating Technology to prevent Interdiffusion between Metallic Fuel and Clad Material (금속연료-피복재 상호확산 방지를 위한 크롬 도금법 적용 연구)

  • Kim, Jun Hwan;Lee, Kang Soo;Yang, Seong Woo;Lee, Byoung Oon;Lee, Chan Bock
    • Korean Journal of Metals and Materials
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    • v.49 no.12
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    • pp.937-944
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    • 2011
  • Studies have been carried out in order to reduce fuel-cladding chemical interaction (FCCI) behavior of metallic fuel in sodium-cooled fast reactors (SFR) using an electroplating technique. A $20{\mu}m$ thick Cr layer has been plated by the electrochemical method in the Sargent bath over the HT9 (12Cr-1Mo) clad material and diffusion couple tests of the U-10Zr metallic fuel as well as the rare earth alloy (70Ce-29La) have been conducted. The results show that the Cr plating can prevent FCCI behavior along the fuel-clad interface. However, cracks developed through the thickness during plating, which resulted in the migration of some fuel constituents. Variation of bath temperature, application of pulse current, and post heat treatment have been conducted to control such cracks. We found out that some conditions like the pulse current and the post heat treatment enhanced the layer property by reducing the internal cracks and improving the diffusion couple test.

Spray-dried powder preparation of pumpkin sweet potato hydrolysates and its physicochemical properties (호박고구마 효소 분해물의 분무건조 분말 제조 및 물리화학적 품질특성)

  • Lee, Dae-Hoon;Jang, Jong-Hyun;Hong, Joo-Heon
    • Food Science and Preservation
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    • v.24 no.2
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    • pp.246-253
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    • 2017
  • This study was conducted prepare spray-dried powder using pumpkin sweet potato hydrolysates and examine the physicochemical properties of the powder. The insoluble dietary fiber and soluble dietary fiber of the pumpkin sweet potato treated by enzyme were 4.17% and 2.07%, respectively. The spray-dried pumpkin sweet potato hydrolysates was manufactured via spray-drying with different forming agents: i.e., pectin 0.1%, 0.5%, 1%, and 2.0%. The moisture contents and total starches of the spray-dried powders were approximately 1.68-2.46 and 45.32-46.51%, respectively. The color of the L and a value decreased, and that of the b and ${\Delta}E$ value increased. The particle size and outer topology of the spray-dried powders were $37.17-42.32{\mu}m$, and its shape was generally globular. The water absorption index of the spray-dried powder (1.74-1.91) was lower than that of the freeze-dried powder (2.15). The water solubility index of the spray-dried powder, 80.75-87.61%, was higher than that of the freeze-dried powder (70.47%). The adhesion values of spray-dried powder to epithelial HT-29 cells were 2.66-6.18% of the initial cell counts, whereas freeze-dried powder showed lower adhesive ability (1.79%). The in vitro human digestibility in the spray-dried powder was 70.09% which is very effective in digestion.