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Screening and Extraction Condition of Antiaging Bioactive Substances from Medicinal Plants (각종 약용 식물로부터 노화 억제 관련 생리활성 물질의 탐색 및 추출 조건)

  • Yu, Hyung-Eun;Dela Paz, Leaniza Michella M.;Bae, Young-Joo;Lee, Dae-Hyoung;Park, Jong-Sang;Kwak, Hahn-Shik;Kim, Ha-Kun;Lee, Jong-Soo
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.34 no.8
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    • pp.1136-1142
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    • 2005
  • To develop new anti-aging foods or cosmetics by using antioxidants, SOD activator and elastase inhibitor, both potent anti-aging substances, were screened from various extracts of medicinal Plants and its optimal extraction conditions were investigated. Antioxidant activity has showed the highest in methanol extracts of Prunus persica (seed; 98.0$\%$). Methanol extracts of Morus alba (leave; 41.0$\%$) showed the highest elastase inhibitory activity while Lycium chinense (fruit; 197$\%$) showed the highest activation effect in SOD activity. The Prunus persica extract that exhibited the highest activity was extracted by treatment of Prunus persica powder with methanol at 40$^{\circ}C$ for 18 h and the SOD activity was maximum with extract from Lycium chinense extracted with deionized water at 30$^{\circ}C$ for 12 h. Elastase inhibitory activity of Morus alba was maximally extracted when it was treated with 70$\%$ methanol at 50$^{\circ}C$ for 12 h.

Background $K^+$ channel currents in WEHI-231 cells, immature B lymphocytes

  • Nam, Joo-Hyun;Woo, Ji-Eun;Kim, Tae-Jin;Uhm, Dae-Yong;Kim, Sung-Joon
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.39-39
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    • 2003
  • In our previous study, WEHI-231, an immature B cell line, showed intractable increase in [C $a^{2+}$]$_{c}$ after the B-cell receptor (BCR) ligation and treatment with 2-aminoethoxydiphenylborate (2-APB), which was never observed in Bal-17, a mature B cell line (Nam et al., 2003, FEBS Lett). In this study, a whole cell voltage clamp study revealed a specific expression of a novel type of $K^{+}$ current, namely voltage-independent background-type $K^{+}$ channels (IK-bg), in WEHI-231 cells. IK-bg was dramatically increase by the application of 2-APB (50 $\square$M), which induced severe hyperpolarization of WEHI-231 from -45 ㎷ to -90 ㎷, When dialyzed with $Mg^{2+}$ and ATP-free pipette solution, a spontaneous development of IK-bg and membrane hyperpolarization were observed. IK-bg was insensitive to classical $K^{+}$ channel blockers (TEA, glibenclamide, $Ba^{2+}$(1 mM)), whereas blocked by quinine and quinidine in a voltage-dependent manner ($IC_{50}$/=6~9 $\square$M at +60㎷). Phorbol myrstate, a PKC activator, decreased the amplitude of IK-bg. Extracellular acidification (pH 6.5) slightly inhibited IK-bg. Arachidonic acid, riluzole, or hyposmotic stress could not affect the IK-bg after the full development by the intracellular dialysis with Mg-ATP-free solution. In a cell-attached mode of single channel recording from WEHI231, we found two types of voltage-independent $K^{+}$ channels with unitary conductance of 300 pS and 120 pS, respectively. Both channels showed very short mean open times and their open probabilities were increase by the application of 2-APB. In Bal-17 cells, no such $K^{+}$ current was observed in 50 cells tested. In summary, WEHI-231 immature B cells express background $K^{+}$ channels. The pharmacological properties and the large unitary conductance suggest that novel types of two-pore domain $K^{+}$ channels (2-P-K channels) might be expressed in WEHI-231, which may provide an intriguing targets of signal transduction in the immature B lymphocytes.e B lymphocytes.

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Inhibition of L-type Ca2+ current by ginsenoside Rd in rat ventricular myocytes

  • Lu, Cheng;Sun, Zhijun;Wang, Line
    • Journal of Ginseng Research
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    • v.39 no.2
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    • pp.169-177
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    • 2015
  • Background: Ginsenoside Rd (GSRd), one of the most abundant ingredients of Panax ginseng, protects the heart via multiple mechanisms including the inhibition of $Ca^{2+}$ influx.We intended to explore the effects of GSRd on L-type $Ca^{2+}$ current ($I_{Ca,L}$) and define the mechanism of the suppression of $I_{Ca,L}$ by GSRd. Methods: Perforated-patch recording and whole-cell voltage clamp techniques were applied in isolated rat ventricular myocytes. Results: (1) GSRd reduced $I_{Ca,L}$ peak amplitude in a concentration-dependent manner [half-maximal inhibitory concentration $(IC_{50})=32.4{\pm}7.1{\mu}mol/L$] and up-shifted the current-voltage (I-V) curve. (2) GSRd ($30{\mu}mol/L$) significantly changed the steady-state activation curve of $I_{Ca,L}$ ($V_{0.5}:-19.12{\pm}0.68$ vs. $-6.26{\pm}0.38mV$; n = 5, p < 0.05) and slowed down the recovery of $I_{Ca,L}$ from inactivation [the time content (${\zeta}$) from 91 ms to 136 ms, n = 5, p < 0.01]. (3) A more significant inhibitive effect of GSRd ($100{\mu}mol/L$) was identified in perforated-patch recording when compared with whole-cell recording [$65.7{\pm}3.2%$ (n = 10) vs. $31.4{\pm}5.2%$ (n = 5), p < 0.01]. (4) Pertussis toxin ($G_i$ protein inhibitor) completely abolished the $I_{Ca,L}$ inhibition induced by GSRd. There was a significant difference in inhibition potency between the two cyclic adenosine monophosphate elevating agents (isoprenaline and forskolin) prestimulation [$55{\pm}7.8%$ (n = 5) vs. $17.2{\pm}3.5%$ (n = 5), p < 0.01]. (5) 1H-[1,2,4]Oxadiazolo[4,3-a]-quinoxalin-1-one (a guanylate cyclase inhibitor) and N-acetyl-$\small{L}$-cysteine (a nitric oxide scavenger) partly recovered the $I_{Ca,L}$ inhibition induced by GSRd. (6) Phorbol-12-myristate-13-acetate (a protein kinase C activator) and GF109203X (a protein kinase C inhibitor) did not contribute to the inhibition of GSRd. Conclusion: These findings suggest that GSRd could inhibit $I_{Ca,L}$ through pertussis toxin-sensitive G protein ($G_i$) and a nitric oxide-cyclic guanosine monophosphate-dependent mechanism.

Immunological benefits by ginseng through reciprocal regulation of Th17 and Treg cells during cyclosporine-induced immunosuppression

  • Heo, Seong Beom;Lim, Sun Woo;Jhun, Joo Yeon;Cho, Mi La;Chung, Byung Ha;Yang, Chul Woo
    • Journal of Ginseng Research
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    • v.40 no.1
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    • pp.18-27
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    • 2016
  • Background: It is not clear whether ginseng affects cyclosporine A (CsA)-induced desirable immunosuppressive action. In this study, we evaluated the immunological influence of combined treatment of ginseng with CsA. Methods: Using CD4+ T cells from mouse spleens stimulated with the T cell receptor (TCR) or allogeneic antigen-presenting cells (APCs), we examined the differentiation of naïve T cells into T helper 1 (Th1), Th2, Th17, and regulatory T cells (Tregs), and their cytokine production during treatment by Korean Red Ginseng extract (KRGE) and/or CsA. The influence of KRGE on the allogeneic T cell response was evaluated by mixed lymphocyte reaction (MLR). We also evaluated whether signal transducer and activator of transcription 3 (STAT3) and STAT5 are implicated in this regulation. Results: Under TCR stimulation, KRGE treatment did not affect the population of CD4+interferon gamma ($IFN{\gamma}$)+ and CD4+interleukin (IL)-4+ cells and their cytokine production compared with CsA alone. Under the Th17-polarizing condition, KRGE significantly reduced the number of CD4+IL-17+ cells and CD4+/phosphorylated STAT3 (p-STAT3)+ cells, but increased the number of CD4+CD25+forkhead box P3 (Foxp3)+ cells and CD4+/p-STAT5+ cells compared with CsA alone. In allogeneic APCs-stimulated CD4+ T cells, KRGE significantly decreased total allogeneic T cell proliferation. Consistent with the effects of TCR stimulation, KRGE reduced the number of CD4+IL-17+ cells and increased the number of CD4+CD25+Foxp3+ cells under the Th17-polarizing condition. Conclusion: KRGE has immunological benefits through the reciprocal regulation of Th17 and Treg cells during CsA-induced immunosuppression.

Characterization of geopolymer made of municipal solid waste incineration ash slag (도시쓰레기 소각재 슬래그로 제조된 지오폴리머의 특성)

  • Kim, Yongsung;Kang, Seunggu
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.24 no.1
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    • pp.15-20
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    • 2014
  • In this research, the geopolymer was fabricated using municipal solid waste incineration ash (denoted as MSWIA) slag and alkali activator, NaOH and its properties were analyzed. Particularly, the effects of NaOH molarity, particle size of MSWIA, and liquid/solids ratio on the compressive strength of geopolymers were investigated. The compressive strength of geopolymers fabricated increased with finer grain size of MSWIA, and optimum value of the liquid/solids ratio was identified as 0.13. As the molarity of the NaOH increased, the compressive strength of geopolymers was increased. Even more the 20 M of NaOH, but the strength was not increased. The calcium aluminum silicate and calcium aluminum silicate hydrate zeolites were generated in the geopolymer fabricated with more than 20 M of NaOH, with some unreacted silica and unknown crystals remained. The highest compressive strength, 163 MPa, of geopolymer was appeared at conditions of curing temperature $70^{\circ}C$, and 20 M of NaOH, indicating that the high concentration of NaOH accelerates the geopolymer reaction and dense microstructure. The high-strength geopolymer fabricated in the present study is expected to contribute significantly to develop the field of cement alternative substances and to improve the recycling rate of MSWIA slag.

Characteristics of Natural Loess (Hwangtoh) Paste Subjected to Geopolymerization (Geopolymerization을 적용한 천연황토 페이스트의 특성)

  • Kim, Baek-Joong;Choi, Hee-Bok;Kang, Kyung-In;Yi, Chong-Ku
    • Journal of the Korea Concrete Institute
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    • v.23 no.1
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    • pp.121-127
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    • 2011
  • In this study, possible use of indigenous natural loess (Hwangtoh) as a new binding material via geopolymerization process is examined. Hwangtoh pastes with four different mix proportions of varying alkali liquid concentrations (6 M, 8 M) and the constituents of the binder as well as the alkali liquid at a constant liquid-to-binder ratio of 0.55 were prepared. Analysis of the natural loess (Hwangtoh) paste was carried out as follows : 1) Measurement of compressive strength and weight of cubic specimens versus curing time; 2) Analysis by X-ray diffraction (XRD) and scanning electron microscope (SEM) about reaction product; 3) Porosity analysis of hardened Hwangtoh paste. The result showed that it is possible to prepare Hwangtoh paste with 29.1 MPa at the age of 7 day by using alkali solution (made as 1 : 4.5 the mass ratio of liquefied $Na_2SiO_3$ and NaOH solution and applying the curing temperature of $60^{\circ}C$). Compressive strength development with respect to the degree of moisture evaporation from the paste seems to be independent of curing temperature. Therefore, it seems that higher early strength of the paste specimens cured at higher temperature can be attributed to both higher rate of reaction and moisture evaporation.

Tricho-dento-osseous Syndrome Mutant Dlx3 Shows Lower Transactivation Potential but Has Longer Half-life than Wild-type Dlx3

  • Cha, Ji-Hun;Ryoo, Hyun-Mo;Woo, Kyung-Mi;Kim, Gwan-Shik;Baek, Jeong-Hwa
    • International Journal of Oral Biology
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    • v.32 no.4
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    • pp.119-125
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    • 2007
  • Dlx3 is a homeodomain protein and is known to play a role in development and differentiation of many tissues. Deletion of four base pairs in DLX3 (NT3198) is causally related to tricho-dento-osseous (TDO) syndrome (OMIM #190320), a genetic disorder manifested by taurodontism, hair abnormalities, and increased bone density in the cranium. The molecular mechanisms that explain the phenotypic characteristics of TDO syndrome have not been clearly determined. In this study, we examined phenotypic characteristics of wild type DLX3(wtDlx3) and 4-BP DEL DLX3 (TDO mtDlx3) in C2C12 cells. To investigate how wtDlx3 and TDO mtDlx3 differentially regulate osteoblastic differentiation, reporter assays were performed by using luciferase reporters containing the promoters of alkaline phosphatase, bone sialoprotein or osteocalcin. Both wtDlx3 and TDO mtDlx3 enhanced significantly all the reporter activities but the effect of mtDlx3 was much weaker than that of wtDlx3. In spite of these differences in reporter activity, electrophoretic mobility shift assay showed that both wtDlx3 and TDO mtDlx3 formed similar amounts of DNA binding complexes with Dlx3 binding consensus sequence or with ALP promoter oligonucleotide bearing the Dlx3 binding core sequence. TDO mtDlx3 exhibits a longer half-life than wtDlx3 and it corresponds to PESTfind analysis result showing that potential PEST sequence was missed in carboxy terminal of TDO mtDlx3. In addition, co-immunoprecipitation demonstrated that TDO mtDlx3 binds to Msx2 more strongly than wtDlx3. Taken together, though TDO mtDlx3 acted as a weaker transcriptional activator than wtDlx3 in osteoblastic cells, there is possibility that during in vivo osteoblast differentiation TDO mtDlx3 may antagonize transcriptional repressor activity of Msx2 more effectively and for longer period than wtDlx3, resulting in enhancement of osteoblast differentiation.

Anti-inflammatory effect of (-)-epigallocatechin-3-gallate on Porphyromonas gingivalis lipopolysaccharide-stimulated fibroblasts and stem cells derived from human periodontal ligament

  • Jung, Im-Hee;Lee, Dong-Eun;Yun, Jeong-Ho;Cho, Ah-Ran;Kim, Chang-Sung;You, Yoon-Jeong;Kim, Sung-Jo;Choi, Seong-Ho
    • Journal of Periodontal and Implant Science
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    • v.42 no.6
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    • pp.185-195
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    • 2012
  • Purpose: (-)-epigallocatechin-3-gallate (EGCG) has been reported to exert anti-inflammatory and antibacterial effects in periodontitis. However, its exact mechanism of action has yet to be determined. The present in vitro study evaluated the anti-in-flammatory effects of EGCG on human periodontal ligament fibroblasts (hPDLFs) and human periodontal ligament stem cells (hPDLSCs) affected by bacterial lipopolysaccharide (LPS) extracted from Porphyromonas gingivalis. Methods: hPDLFs and hPDLSCs were extracted from healthy young adults and were treated with EGCG and/or P. gingivalis LPS. After 1, 3, 5, and 7 days from treatment, cytotoxic and proliferative effects were evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and bromodeoxyuridine assay, respectively. And then, the gene expressions of hPDLFs and hPDLSCs were observed for interleukin (IL)-$1{\beta}$, IL-6, tumor necrosis factor (TNF)-${\alpha}$, osteoprotegerin (OPG), receptor activator of nuclear factor kappa-B ligand (RANKL), and RANKL/OPG using real-time polymerase chain reaction (PCR) at 0, 6, 24, and 48 hours after treatment. The experiments were performed with the following groups for hPDLFs and hPDLSCs; 1) No treat, 2) EGCG alone, 3) P. gingivalis LPS alone, 4) EGCG+P. gingivalis LPS. Results: The 20 ${\mu}M$ of EGCG and 20 ${\mu}g/mL$ of P. gingivalis LPS had the lowest cytotoxic effects, so those concentrations were used for further experiments. The proliferations of hPDLFs and hPDLSCs increased in all groups, though the 'EGCG alone' showed less increase. In real-time PCR, the hPDLFs and hPDLSCs of 'EGCG alone' showed similar gene expressions to those cells of 'no treat'. The gene expressions of 'P. gingivalis LPS alone' in both hPDLFs and hPDLSCs were highly increased at 6 hours for IL-$1{\beta}$, IL-6, TNF-${\alpha}$, RANKL, and RANKL/OPG, except the RANKL/OPG in hPDLSCs. However, those increased gene expressions were down-regulated in 'EGCG+P. gingivalis LPS' by the additional treatment of EGCG. Conclusions: Our results demonstrate that EGCG could exert an anti-inflammatory effect in hPDLFs and hPDLSCs against a major pathogen of periodontitis, P. gingivalis LPS.

Korean Mistletoe Viscum album coloratum Induces Specific and Non-Specific Immune Responses in Japanese Eel Anguilla japonica

  • Choi, Sang-Hoon;Rhow, Jin-Goo;Choi, Min-Soon;Park, Sung-Woo;Park, Kwan-Ha;Yoon, Jong-Man;Park, Gyung-Il
    • Journal of Aquaculture
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    • v.22 no.1
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    • pp.56-62
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    • 2009
  • Effects of Korean mistletoe extracts (KM-110), Viscum album coloratum on the specific and non-specific immune responses of Japanese eel Anguilla japonica were examined. The optimal concentration not showing toxicity of KM-110 was determined to $30-40{\mu}g/ml$ in vitro and $100{\mu}g$/100 g of fish in vivo. Even $1000{\mu}g$ of KM-110/100 g of fish did not show any clinical problem in fish though the levels of toxic parameters were slightly increased. In terms of antibody production, KM-110 significantly elicited more antibody production than FCA or $\beta$-glucan. $\beta$-glucan plus KM-110 group synergistically enhanced antibody production. There was no significant difference between KM-110 and KM-110 plus $\beta$-glucan group. The ROI production by head kidney (HK) leucocytes of eel injected with 500 or $1000{\mu}g$ KM-110 was significantly (P<0.05) enhanced than the control and FCA-treated group. Maximum increase in the NBT reduction value was observed in $1000{\mu}g$ KM-110 group but no significant difference was found between 500 and $1000{\mu}g$ KM group. The level of serum lysozyme activity was significantly (P<0.05) higher in the 500 and $1000{\mu}g$ KM-110- or FCA-treated group than in the control and $200{\mu}g$ KM-110 group. The phagocytic activities of HK leucocytes isolated from eel injected with 500 and $1000{\mu}g$ KM-110 were significantly (P<0.05) higher than $200{\mu}g$ KM-110 and PBS-injected control group. Korean mistletoe appeared to be a good activator of the specific and non-specific immune responses of Japanese eel.

Luminescence Characteristics of ${Y_2}{O_3}$:Eu Phosphor Treated with $\alpha$-${Fe_2}{O_3}$Prepared by Two Different Methods Using $FeSO_4$.$7H_2$O ($FeSO_4$.$7H_2$O를 이용하여 서로 다른 방법으로 만들어진 $\alpha$-${Fe_2}{O_3}$를 표면처리한 ${Y_2}{O_3}$:Eu 적색 형광체의 발광 특성)

  • 김봉철;이춘엽;송윤호;서경수;이진호;이남양;김동국;박이순;이병교
    • Journal of the Korean Ceramic Society
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    • v.38 no.12
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    • pp.1115-1122
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    • 2001
  • The tendency of the miximum brightness of $Y_2$O$_3$:Eu phosphor with various activator concentration, by different surface treatment methods as well as different exciting energies were investigated. The surface treatment methods were the adsorption method used $\alpha$-Fe$_2$O$_3$powder prepared emulsion-drying process and the precipitation method used FeSO$_4$/ethanol. Eu concentration of maximum brightness of $Y_2$O$_3$:Eu phosphor prepared by solid-solid state was changed with various exciting energies. The concentrations were 0.02 mol at VUV(147 nm) as well as 400 V and 0.03 mol at 5 kV. The phosphor treated both by adsorption method and precipitation method showed decreasing luminescent intensity with increasing amount of $\alpha$-Fe$_2$O$_3$, and the methods are chosen by exciting energy. Adsorption method was effective in a low voltage and VUV(147nm) region, and precipitation method was effective in the high voltage region.

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