• Title/Summary/Keyword: MEK1/2

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Induction of Dectin-1 Expression and Intracellular Signal Transduction by β-Glucan of Ganoderma lucidum (불로초의 β-Glucan에 의한 Dectin-1 발현 유도와 세포 내 신호전달)

  • Ryu, Han Wook;Kim, Ha Won
    • The Korean Journal of Mycology
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    • v.46 no.2
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    • pp.161-176
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    • 2018
  • Fungal ${\beta}$-glucan, known to have immunostimulatory and antitumor activities, can be recognized by host immune cells as one of the pathogen-associated molecular patterns (PAMPs). Although there are several reports on the diverse immunostimulatory activities of ${\beta}$-glucan, little is known about the intracellular signal transduction of ${\beta}$-glucan. Stimulation of RAW264.7 macrophage cells with ${\beta}$-glucan from Ganoderma lucidum induced the expressions of dectin-1, toll-like receptor 2 (TLR2), TLR4, and TLR6 at the transcription stage. Treatment with ${\beta}$-glucan also induced inflammatory mediators such as macrophage inflammatory proteins (MIP)-$1{\alpha}$, MIP-$1{\beta}$, MIP-$1{\gamma}$, interleukin (IL)-$1{\beta}$, and tumor necrosis factor (TNF)-${\alpha}$. Treatment of the cells with polymyxin B, an inhibitor of lipopolysaccharides (LPS), blocked the induction of inflammatory mediators in LPS- or ${\beta}$-glucan-stimulated systems. Pretreatment of the cells in our cell culture system with LY294002, a phosphoinositide 3-kinase (PI3K) inhibitor, or U0126, a mitogen-activated protein kinase/extracellular-signal-regulated kinase (MAPK/ERK) kinase (MEK)1/MEK2 inhibitor, led to a reduction in the induction of inflammatory mediators in a concentration-dependent manner. These results show that stimulation of the macrophage cells by ${\beta}$-glucan induced the expressions of both dectin-1 and TLRs. We also found that the PI3K/Akt and MEK pathways were involved in the induction of inflammatory mediators in macrophage cells during intracellular signal transduction of ${\beta}$-glucan.

The Inhibitory Effect of Broccoli in Cruciferous Vegetables Derived-Sulforaphane on Vascular Tension (브로콜리 유래 Sulforaphane의 혈관 수축성 조절 효과)

  • Je, Hyun Dong
    • YAKHAK HOEJI
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    • v.58 no.4
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    • pp.223-228
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    • 2014
  • The present study was undertaken to investigate the influence of sulforaphane on vascular smooth muscle contractility and to determine the mechanism involved. We hypothesized that sulforaphane, the primary ingredient of broccoli of cruciferous vegetables, plays a role in vascular relaxation through inhibition of Rho-kinase in rat aortae. Intact of denuded arterial rings from male Sprague-Dawley rats were used and isometric tensions were recorded using a computerized data acquisition system. Interestingly, sulforaphane significantly inhibited fluoride, phorbol ester or thromboxane $A_2$ mimetic-induced contraction in denuded muscles suggesting that additional pathways different from endothelial nitric oxide synthesis such as inhibition of Rho-kinase or MEK might be involved in the vasorelaxation. Furthermore, sulforaphane inhibited thromboxane $A_2$-induced increases in pERK1/2 levels suggesting the mechanism including inhibition of thromboxane $A_2$-induced increases in ERK1/2 phosphorylation. This study provides evidence that sulforaphane induces vascular relaxation through inhibition of Rho-kinase or MEK in rat aortae.

A Study on the Way of Pulse-diagnosis by Dongeuibogam (동의보감(東醫寶鑑)중 맥법(脈法)에 관한 연구(硏究))

  • Joo, Shin-Tak;Kim, Jung-Guk;Park, Won-Hwan
    • The Journal of the Society of Korean Medicine Diagnostics
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    • v.15 no.1
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    • pp.1-28
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    • 2011
  • Objectives: A study on the importance and consistency of pulse-daignosis in the Dongeuibogam Methods: We used Deyeuk Dongeuibogam of Dongeuibogam publishing company from original photographic edition. Results: The frequency of 27 mek (pulse condition) in Dongeuibogam is as in the following. Bumek (Floating pulse) appeared 120(8.9%) times, wanmek (moderate pulse) appeared 28(2%) times, chokmek (running pulse) appeared 7 (0.5%) times, gyumek (hollow pulse) appeared 19 (1.4%) times, saekmek (uneven pulse) appeared 33 (2.4%) times, sapmek (uneven pulse) appeared 51 (3.8%) times, kyulmek (knotted pulse) appeared 18 (1.3%) times, whalmek (slippery pulse) appeared 69 (5.1%) times, chimek (slow pulse) appeared 43 (3.2%) times, demek (intermittent pulse) appeared 13 (1%) times, silmek (replete pulse) appeared 45 (3.3%) times, bokmek (deep-sited pulse) appeared 29 (2.1%) times, neomek (firm pulse) appeared 4 (0.3%) times, hyunmek (taut pulse) appeared 110 (8.1%) times, yumek (soft pulse) appeared 20 (1.5%) times, dongmek (short and rapid pulse) appeared 16 (1.2%) times, kinmek (tense pulse) appeared 67 (5%) times, yakmek (weak pulse) appeared 46 (3.4%) times, semek (thready pulse) appeared 62 (4.6%) times, hongmek (full pulse) appeared 50 (3.7%) times, jangmek (long pulse) appeared 14 (1%) times, sakmek (rapid pulse) appeared 103 (7.6%) times, mimek (indistinctive pulse) appeared 65 (4.8%) times, danmek (short pulse) appeared 16 (1.2%) times, demek (large pulse) appeared 106 (7.9%) times, chimmek (deep pulse) appeared 112 (8.3%) times, heomek appeared 70 (5.2%) times, sanmek (scattered pulse) appeared 14(1%)times. Conclusions: We can know Donguibogam is given on the basis 27mek (pulse condition), because the frequency of 27mek (pulse condition) is high. But there are another expressions. So we can not say that Donguibogam is consistent in expressing mekbub(the way of pulse-diagnosis).

Expression of MEK1 Fusion Protein in Yeast for Developing Cell Based Assay System, a Major Substrate of LeTx (Yeast내에서 MEK1 융합 단백질 발현 및 Lethal Factor 활성 검증)

  • Hwang, Hye-Hyun;Kim, Joung-Mok;Choi, Kyoung-Jae;Park, Hae-Chul;Han, Sung-Hwan;Chung, Hoe-Il;Koo, Bon-Sung;Park, Joon-Shik;Yoon, Moon-Young
    • Korean Journal of Microbiology
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    • v.42 no.3
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    • pp.195-198
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    • 2006
  • Lethal toxin is a critical virulence factor of anthrax. It is composed two protein: protective antigen (PA) and lethal factor (LF). PA binds to specific cell surface receptors and, forms a membrane channel that mediates entry of LF into the cell. LF is a zinc-dependent metalloprotease, which cleaves MKKs [MAPK (mitogen-activated protein kinase) kinases] at peptide bonds very close to their N-termini. In this study, we suggest application of cell-based assays in the early phase of drug discovery, with a particular focus on the use of yeast cells. We constructed MEK1 expression system in yeast to determine LF activity and approached cell-based assay system to screen inhibitors, in which the results covering the construction of LF-substrate in yeast expression vector, expression, and LF-mediated proteolysis of substrate were described. These results could provided the basic steps in design of cell-based assay system with the high efficiency, rapidly and easy way to screening of inhibitors.

Effect of retinoic acid and delta-like 1 homologue (DLK1) on differentiation in neuroblastoma

  • Kim, Yu-Ri
    • Nutrition Research and Practice
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    • v.4 no.4
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    • pp.276-282
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    • 2010
  • The principal objective of this study was to evaluate the chemopreventive and therapeutic effects of a combination of all-trans-retinoic acid (RA) and knockdown of delta-like 1 homologue (Drosophila) (DLK1) on neuroblastoma, the most common malignant disease in children. As unfavorable neuroblastoma is poorly differentiated, neuroblastoma cell was induced differentiation by RA or DLK1 knockdown. Neuroblastoma cells showed elongated neurite growth, a hallmark of neuronal differentiation at various doses of RA, as well as by DLK1 knockdown. In order to determine whether or not a combination of RA and DLK1 knockdown exerts a greater chemotherapeutic effect on neuroblastoma, cells were incubated at 10 nM RA after being transfected with SiRNA-DLK1. Neuronal differentiation was increased more by a combination of RA and DLK1 knockdown than by single treatment. Additionally, in order to assess the signal pathway of neuroblastoma differentiation induced by RA and DLK1 knockdown, treatment with the specific MEK/ERK inhibitors, U0126 and PD 98059, was applied to differentiated neuroblastoma cells. Differentiation induced by RA and DLK1 knockdown increased ERK phosphorylation. The MEK/ERK inhibitor U0126 completely inhibited neuronal differentiation induced by both RA and DLK1 knockdown, whereas PD98059 partially blocked neuronal differentiation. After the withdrawal of inhibitors, cellular differentiation was fully recovered. This study is, to the best of our knowledge, the first to demonstrate that the specific inhibitors of the MEK/ERK pathway, U0126 and PD98059, exert differential effects on the ERK phosphorylation induced by RA or DLK1 knockdown. Based on the observations of this study, it can be concluded that a combination of RA and DLK1 knockdown increases neuronal differentiation for the control of the malignant growth of human neuroblastomas, and also that both MEK1 and MEK2 are required for the differentiation induced by RA and DLK1 knockdown.

Urinary Metabolites of Dimethylformamide, Methyl Ethyl Ketone, and Toluene exposed Workers in Synthetic Leather Factories (일부 합성피혁 근로자들의 Dimethylformamide, Methyl Ethyl Ketone, Toluene 노출에 따른 요중 대사물질)

  • Choi, Ho-Chun;Kim, Kang Yoon;An, Sun-Hee;Lee, Young-Ja;Chung, Kyou-Chull
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.11 no.2
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    • pp.135-144
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    • 2001
  • This study was performed to measure airborne dimethylformamide(DMF), methyl ethyl ketone(MEK) and toluene and their urinary metabolites concentrations and to determine the relationship between airborne and urinary concentration. Airborne samples and their urinary metabolites were measured 98 male workers who work for 8 synthetic leather factories in a portion of Kyoung-In area. Urine samples were collected at end-of-shift to estimate the exposure levels. 1. The concentration of airborne DMF by process was 8.81 ppm for wet-mixing, 15.05 ppm for wet-coating, 6.03 ppm for dry-mixing, 5.58 ppm for dry-coating, 5.37 ppm for printing, and 9.03 ppm for total. There was statistically significant difference by process. Urinary NMF concentrations of wet-mixing, wet-coating, dry-mixing, dry-coating and printing were $90.55mg/{\ell}$, $79.80mg/{\ell}$, $39.86mg/{\ell}$, $25.23mg/{\ell}$, and $38.15mg/{\ell}$, respectively, and total geometric mean was $56.24mg/{\ell}$. There was significant difference by process. 2. The concentration of airborne MEK by process was 1.89 ppm for wet-mixing, 1.96 ppm for wet-coating, 10.33 ppm for dry-mixing, 29.24 ppm for dry-coating, 14.98 ppm for printing, and 4.87 ppm for total. There was statistically significant difference by process. Urinary MEK concentrations of wetmixing, wet-coating, dry-mixing, dry-coating and printing were $0.93mg/{\ell}$, $0.70mg/{\ell}$, $3.29mg/{\ell}$, $3.29mg/{\ell}$, and $1.06mg/{\ell}$, respectively, and total geometric mean was $1.25mg/{\ell}$. There was statistically significant difference by process. Urinary MEK 3. The concentration of airborne toluene by process was 0.35ppm for wet-mixing, 0.42ppm for wet-coating, 2.95ppm for dry-mixing, 11.67ppm for dry-coating, 4.88ppm for printing, 1.24ppm for total. There was statistically significant difference by process. Urinary hippuric acid concentrations of wet-mixing, wet-coating, dry-mixing, dry-coating and printing were 0.24g/g creatinine, 0.21g/g creatinine, 0.34g/g creatinine, 0.52g/g creatinine, and 0.29g/g creatinine, respctively and total geometric mean was 0.28g/g creatinine. There was statistically significant difference by process. 4. No. of exceeded KPEL was 40 workers(40.8%) for DMF(10ppm), 1 worker(1.0%) for MEK(200ppm), and no worker for toluene(100ppm). No. of exceeded KBEI was 62 workers(63.3%) for urinary NMF($40mg/{\ell}$), 29 workers(29.6%) for urinary MEK, 1 worker(1.0%) for urinary hippuric acid. 5. The regression equations were Log(NMF)=0.4094*Log(DMF)+1.3587(r=0.4516) for DMF, Log(MEKU)=0.1859*Log(MEK)-0.0324(r=0.3303) for MEK, Log(HA)=0.2106*Log(Toluene)-0.5685(r=0.4497) for toluene. Synthetic leather factory workers expose to 3 kinds of organic solvents which are DMF, MEK and toluene. Their urinary NMF and MEK levels were higher than their concentration levels through respiratory. It seems that the urinary levels were affected skin absorption for working habit and alcohol intake.

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The effect of dilution solvent ratio on dewaxing of waxy oils (Waxy Oil 탈납에 있어서 용제희석의 영향)

  • 김주항
    • Tribology and Lubricants
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    • v.3 no.2
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    • pp.34-43
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    • 1987
  • The effect of dilution solvent ratio, and the mixing ratio of MEK and toluene on the solvent dewaxing process has been studied. The results of this study are Summarized as follows; 1. The best mixing ratio of solvent of MEK process and Toluene when in case of light and medium Waxy oils, which has low viscosity was 48: 52, and when in case of heavy waxy oil, which has high viscosity was 45: 55. 2. The best dilution solvent ratio when in case of low viscosity oils. was 1:2.8 against waxy oils after annexing 5 times divided as well as, when incase of high viscosity oil was 1:3.5 after annexing 2 times divided. 3. The chilling temperature was -26$\circ$C and the reguired dewaxing time depends on the viscosity of waxy oils.

A Convenient Method on the Methyl-Ethyl-Ketone Extraction of $^{99m}TcO^-{_4}$ ($^{99m}TcO^-{_4}$의 메틸-에틸-케톤-간편 추출법)

  • Lee, Jong-Du;Lee, Byung-Hyn
    • Journal of Radiation Protection and Research
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    • v.9 no.2
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    • pp.103-111
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    • 1984
  • A convenient method of $^{99m}Tc$-methyl-ethyl-ketone (MEK) extraction technique was developed and a mobile $^{99m}Tc$-extraction generator was designed. The MEK extraction and the phase separation of $^{99m}TcO^-{_4}$ were carried out with a simple procedure in the same container. The shielding of $^{99}Mo$ radioactivity was made with one lead container. The system was simplified by shielding $^{99m}TcO_4{^-}({\gamma}_e=0.14\;MeV)$ separately. $^{99m}TcO^-{_4}\;in\;^{99m}Tc-MEK$ extract was recovered by adsorption and elution only, and therefore, the possibility of volatilization was reduced. The volume of $^{99m}TcO^{-}{_4}$-saline product was reduced to 1 ml by using a small alumina column and the column operation time was shortened. The separation time of $^{99m}Tc$ was reduced to 30 minutes, and the operation was carried out at the outside of the shielding. The system was designed to operate under the condition of bacteria-free.

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Manufacture of the Hydrophobic HY-type Zeolite-honeycomb and Its Adsorption/Desorption Characteristics for the Benzene, o-xylene, and MEK (소수성 HY-형 제올라이트제 하니컴의 제조 및 그 하니컴의 벤젠, o-xylene, MEK에 대한 흡.탈착특성)

  • Mo, Se-Young;Jeon, Dong-Hwan;Kwon, Ki-Seung;Sohn, Jong-Ryeul
    • Journal of Korean Society for Atmospheric Environment
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    • v.23 no.1
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    • pp.84-96
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    • 2007
  • We performed the experiments to manufacture the hydrophobic $200cells/in^2$-zeolite honeycomb using HY-type zeolite of Si/Al ratio of 80 for separating and removing the VOCs emitted from small and medium size-plants by adsorption and to determine the drying method for the honeycomb at $105^{\circ}C$ without cracking, then measured performances of the honeycomb to adsorb the benzene, o-xylene, and MEK and to desorb the benzene and MEK saturated on the honeycomb by the nitrogen gas as the desorption gas. As a results, the good honeycomb was formed and the honeycomb was not cracked when the mixing ratio of the zeolite to bentonite to methyl cellulose to polyvinyl alcohol to glycerine to water is 100 : 8.73 : 2.18 : 4.19 : 1.38 : 126 and dried the honeycomb at $105^{\circ}C$ for 24 hours in the drying oven. The shape of the dried honeycomb was not changed after calcination, and the compressive strengths of the honeycomb after drying and calcination were 6.7 and $0.69kg/cm^2$, respectively. The adsorption efficiencies of the honeycomb for benzene, o-xylene, and MEK were $92{\sim}96%$ at the room temperature. The desorption efficiency at $180^{\circ}C$ was higher than that at $150^{\circ}C\;by\;1.5{\sim}13.8%$ depending on the flow rate of the nitrogen gas, and it was found that desorption efficiency is higher than 85% at $180^{\circ}C$ and 1.0L/min of the nitrogen gas. At $180^{\circ}C$ and 0.2 L/min, the concentration of the benzene and MEK in the used desorption gas are higher than 40,000 and 50,000ppm, respectively, so it be used as the fuel for preheating the desorption gas fed into the column in desorption cycle.

Radiosynthesis of $[^{11}C]6-OH-BTA-1$ in Different Media and Confirmation of Reaction By-products. ($[^{11}C]6-OH-BTA-1$ 조제 시 생성되는 부산물 규명과 반응용매에 따른 표지 효율 비교)

  • Lee, Hak-Jeong;Jeong, Jae-Min;Lee, Yun-Sang;Kim, Hyung-Woo;Lee, Eun-Kyoung;Lee, Dong-Soo;Chung, June-Key;Lee, Myung-Chul
    • Nuclear Medicine and Molecular Imaging
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    • v.41 no.3
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    • pp.241-246
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    • 2007
  • Purpose: $[^{11}C]6-OH-BTA-1$ ([N-methyl-$^{11}C$]2-(4'-methylaminophenyl)-6-hydroxybenzothiazole, 1), a -amyloid imaging agent for the diagnosis of Alzheimer's disease in PET, can be labeled with higher yield by a simple loop method. During the synthesis of $[^{11}C]1$, we found the formation of by-products in various solvents, e.g., methylethylketone (MEK), cyclohexanone (CHO), diethylketone (DEK), and dimethylformamide (DMF). Materials and Methods: In Automated radiosynthesis module, 1 mg of 4-aminophenyl-6-hydroxybenzothiazole (4) in 100 l of each solvent was reacted with $[^{11}C]methyl$ triflate in HPLC loop at room temperature (RT). The reaction mixture was separated by semi-preparative HPLC. Aliquots eluted at 14.4, 16.3 and 17.6 min were collected and analyzed by analytical HPLC and LC/MS spectrometer. Results: The labeling efficiencies of $[^{11}C]1$ were $86.0{\pm}5.5%$, $59.7{\pm}2.4%$, $29.9{\pm}1.8%$, and $7.6{\pm}0.5%$ in MEK, CHO, DEK and DMF, respectively. The LC/MS spectra of three products eluted at 14.4, 16.3 and 17.6 mins showed m/z peaks at 257.3 (M+1), 257.3 (M+1) and 271.3 (M+1), respectively, indicating their structures as 1, 2-(4'-aminophenyl)-6-methoxybenzothiazole (2) and by-product (3), respectively. Ratios of labeling efficiencies for the three products $([^{11}C]1:[^{11}C]2:[^{11}C]3)$ were $86.0{\pm}5.5%:5.0{\pm}3.4%:1.5{\pm}1.3%$ in MEK, $59.7{\pm}2.4%:4.7{\pm}3.2%:1.3{\pm}0.5%$ in CHO, $9.9{\pm}1.8%:2.0{\pm}0.7%:0.3{\pm}0.1%$ in DEK and $7.6{\pm}0.5%:0.0%:0.0%$ in DMF, respectively. Conclusion: The labeling efficiency of $[^{11}C]1$ was the highest when MEK was used as a reaction solvent. As results of mass spectrometry, 1 and 2 were conformed. 3 was presumed.