• Title/Summary/Keyword: MEK1/2

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Semaphoring mAb: a New Guide in RIT in Inhibiting the Proliferation of Human Skin Carcinoma

  • Liu, Yuan;Ma, Jing-Yue;Luo, Su-Ju;Sun, Chen-Wei;Shao, Li-Li;Liu, Quan-Zhong
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.3
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    • pp.941-945
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    • 2015
  • Semaphoring is a transmembrane receptor which participates in many cytokine-mediated signal pathways that are closely related to the angiogenesis, occurrence and development of carcinoma. The present study was designed to access the effect of mono-antibody (mAb) guided radioimmunotherapy (RIT) on skin carcinoma and investigate the potential mechanisms. Semaphoring mAb was acquired from mice (Balb/c), purified with rProtein A column; purity, concentration and activity were tested with SDS-PAGE and indirect ELISA; specificity and expression on the cutanuem carcinoma line and tissue were tested by Western blotting; morphology change was assessed by microscopy. MTT assay and colony inhibition tests were carried out to test the influence on the proliferation of tumor cells; Western blotting was also carried out for expression of apoptosis-associated (caspase-3, Bax, Bcl-2) and proliferation-related (PI3K, p-Akt, Akt, p-ERK1/2, ERK1/2) proteins and analyse the change in signal pathways (PI3K/Akt and MEK/ERK). The purity of purified semaphorin mAb was 96.5% and the titer is about $1{\times}10^6$. Western blotting showed semaphoring mAb to have specifically binding stripes with semaphoring b1b2 protein, B16F10, and A431 cells at 39KDa, 100KDa and 130KDa, respectively. Positive expression was detected both in cutanuem carcinoma line and tissue and it mostly located in cell membranes. MMT assay revealed dose-relate and time-relate inhibitory effect of semaphorin mAb on A431 and B16F10. Colony inhibition tests also showed dose-relate inhibitory effects. Western blotting demonstrated the expression of apoptosis and proliferation-related protein and changes in signal pathway. In conclusion, we demonstrated that semaphorin is highly expressed on the tumor cell-surfaces and RIT with semaphorin mAb has effect in i nhibiting proliferation and accelerating apoptosis of tumor cells.

A Study on Desorption Efficiency for Polar Solvents Collected on Charcoal Tube (활성탄관에 포집된 극성유기용제의 탈착효율에 관한 연구)

  • Kim, Kyeong-Ran;Paik, Nam-Won
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.5 no.1
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    • pp.104-118
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    • 1995
  • This study was performed to evaluate factors affecting desorption of organic solvents collected on charcoal tube and to find out the optimum condition. Desorption efficiency for polar analytes was improved when several polar desorption solvents such as methanol, dimethylformamide(DMF), 2-(2-butoxyethoxy)ethanol were added to carbon disulfide($CS_2$). The best improvement was achieved when 10% dimethylformamide(DMF) in $CS_2$ was used as desorption solvent. During storage of polar analytes, recovery was greatly reduced. Especially, the recovery of cyclohexanone was decreased to 18.1 % after a month storage at $34^{\circ}C$. After two weeks storage, recovery of polar analytes was sharply decreased. Water adsorbed on charcoal interfered the recovery of polar analytes but didn't interfere that one of nonpolar solvent, toluene. When 10% DMF in $CS_2$ was used as desorption solvent, the effect of water on recovery was decreased, comparing with Desorption efficiency increased when analyte loading increased, and usage of 10% DMF in $CS_2$ decreased the loading effect. Increasing volume of desorption solvent was not effective to improve desorption efficiency of analytes when 10% DMF was used. Continuous shaking and sonication is not helpful to increase the desorption efficiency of analytes except cyclohexanone using 10% DMF. When silica gel used as adsorbent, methanol was better desorbent than dimethylsulfoxide. Analytes adsorbed on silica gel showed high recovery in low concentration and less affected by humidity. On the basis of this study, the following conclusions have been drawn. To improve the recovery of polar organic materials in air samples, it is necessary to analyze samples as soon as possible after they were collected. Otherwise, samples must be stored at low temperature. Using two components of desorption solvents, such as 10% DMF in $CS_2$, the effects of loading and humidity decreased for polar analytes such as methyl ethyl ketone and methyl isobutyl ketone. When work place has high humidity with low concentration of polar organic solvents, silica gel can be used as adsorbent, because it produces quantitative recovery for polar analytes at this condition. But it should be noted that high humidity makes breakthrough easy in silica gel samples.

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hEPO 형질전환 돼지 후대의 임신기간내 혈액조성 변화

  • 이연근;이현기;이풍연;김성우;김정호;박진기;장원경
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.38-38
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    • 2003
  • 본 연구는 사람 조혈촉진유전자(hEPO)가 도입된 형질전환돼지 Fl 수컷과 암컷간 교배로 F2 생산시, Fl 모돈에 대한 임신기간(약 114일)내 혈액조성(적혈구, 백혈구, Hemoglobin, Hematocrit value, MCV, MCH, MCHC, 혈소판) 변화를 구명하기 위하여 실시하였다. 시험에 이용된 암퇘지는 형질전환 F1 암컷 2마리와 일반돼지 1마리였고 교배와 동시에 임신기간동안(114일) 5일 간격으로 5ml의 혈액을 경정맥(jugular vein)에서 채취하였으며 혈액조성분석은 Celltac MEK 5108K(Nihon Kohden, Japan)를 이용하였는데 얻어진 결과는 다음과 같다. 임신기간중 혈액내 백혈구의 변화는 일반돼지의 경우 13.8($\pm$2.4)$\times$$10^3$/ul 로임신기간내에 일정한 변화양상을 보였으나 형질전환돼지의 경우 29.4($\pm$19.4)$\times$$10^3$와 22.5($\pm$14.1)$\times$$10^3$/ul 으로 일반돼지보다는 높게 나타났으며 임신기간내에 매우 높은 변화 양상을 나타내었다. 또한 적혈구는 일반돼지(7.2$\pm$0.7 $\times$$10^{6}$/ul)에 비하여 형질전환돼지(11.5$\pm$0.5 와 11.9$\pm$0.5 $\times$$10^{6}$ul)에서 매우 높게 나타났으며 변화양상은 일반돼지와 마찬가지로 작게 나타났다. 적혈구와 함께 Hemoglobin(Hb)과 평균혈구용적율(Hematocrit value ;, HCT) 모두 형질전환돼지가 일반돼지보다 높게 나타났으며 변화양상 또한 작게 나타남으로서 사람 조혈촉진유전자(hEPO)가 형질전환된 돼지에서는 지속적으로 발현되어 조혈촉진이 이루어지고 있으며, 조혈촉진에 의해 헤모글로빈 및 적혈구가 생성되어 혈액내 높은 혈구용적율(HCT)을 이루고 있음을 알 수 있었다. 적혈구 평균용적(Mean Corpuscular Volume ; MCV), 평균적혈구혈색소량(Mean Corpuscular Hemoglobin ; MCH), 평균적혈구혈색소농도(Mean Corpuscular Hemoglobin Concentration ; MCHC) 그리고 혈소판(Platelets) 분석결과 형질전환돼지가 일반돼지보다 약간 높은 수치를 나타냈으며 변화양상 또한 유사한 결과를 나타내었다. 이와 같은 시험분석결과를 토대로 사람 조혈촉진유전자(hEPO)가 형질전환된 돼지는 유즙으로 발현할 수 있도록 형질전환 되었음에도 불구하고 헤모글로빈 및 적혈구가 증가함으로서 형질전환돼지 개체의 혈장으로도 사람 조혈촉진인자가 분비하고 있음을 간접적으로 알 수 있었다. 정상돼지 보다 형질전환돼지가 약 30% 높은HCT 수준을 보였으며 이러한 현상은 사람에서는 적혈구증다증(erythrocytosis)으로 분류되고 있다. 이에 대한 고찰은 형질전환돼지 자체의 생리적 문제점(side effects)에 대한 해결과 더불어 기존의 인간질병에 대한 모델동물로서의 이용 가능성을 제시할 수 있는 것으로 사료된다.

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Panax ginseng and its ginsenosides: potential candidates for the prevention and treatment of chemotherapy-induced side effects

  • Wan, Yan;Wang, Jing;Xu, Jin-feng;Tang, Fei;Chen, Lu;Tan, Yu-zhu;Rao, Chao-long;Ao, Hui;Peng, Cheng
    • Journal of Ginseng Research
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    • v.45 no.6
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    • pp.617-630
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    • 2021
  • Chemotherapy-induced side effects affect the quality of life and efficacy of treatment of cancer patients. Current approaches for treating the side effects of chemotherapy are poorly effective and may cause numerous harmful side effects. Therefore, developing new and effective drugs derived from natural nontoxic compounds for the treatment of chemotherapy-induced side effects is necessary. Experiments in vivo and in vitro indicate that Panax ginseng (PG) and its ginsenosides are undoubtedly non-toxic and effective options for the treatment of chemotherapy-induced side effects, such as nephrotoxicity, hepatotoxicity, cardiotoxicity, immunotoxicity, and hematopoietic inhibition. The mechanism focus on anti-oxidation, anti-inflammation, and anti-apoptosis, as well as the modulation of signaling pathways, such as nuclear factor erythroid-2 related factor 2 (Nrf2)/heme oxygenase-1 (HO-1), P62/keap1/Nrf2, c-jun Nterminal kinase (JNK)/P53/caspase 3, mitogen-activated protein kinase (MEK)/extracellular signal-regulated kinases (ERK), AMP-activated protein kinase (AMPK)/mammalian target of rapamycin (mTOR), mitogen-activated protein kinase kinase 4 (MKK4)/JNK, and phosphatidylinositol 3-kinase (PI3K)/AKT. Since a systemic review of the effect and mechanism of PG and its ginsenosides on chemotherapy-induced side effects has not yet been published, we provide a comprehensive summarization with this aim and shed light on the future research of PG.

Longitudinal Gradients and Seasonal Dynamics of Nutrients, Organic Matter and Conductivity Along the Main Axis of Han-River

  • Kim, Bit-Na;Lee, Sang-Jae;Seo, Jin-Won;An, Kwang-Guk
    • Korean Journal of Ecology and Environment
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    • v.41 no.4
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    • pp.457-465
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    • 2008
  • The purpose of the study was to evaluate spatial and temporal dynamics of nutrients (TN, TP), organic pollution (BOD, COD), and ionic dynamics (electrical conductivity, EC) in the North Han-River, South Han-River, and merged downriver using the dataset of $1998{\sim}2007$, obtained from the MEK (Ministry of Environment, Korea). Accord. ing to interannual nutrient analysis, TN varied slightly in the North Han-River and South Han-River, but decreased in the merged downriver along with BOD. Longitudinal analysis in the water quality showed that BOD, COD, and nutrients had linear decreasing trend along the main axis of headwater-to-downriver. Concentrations of TP and TN in the North Han-River averaged $26.97{\mu}g\;L^{-1}$, $1.696mg\;L^{-1}$, respectively, which were minimum in the three watersheds, followed by South Han-River and then the merged downriver in order. Ratios of TN:TP in the watersheds were >40 in all the sites, indicating that nitrogen may be enough for periphyton or phytoplankton growth and phosphorus may be limited partially. After the North Han-River water is merged with South Han-River, the concentrations of BOD, COD, TN, and TP were similar to the values of $S6{\sim}S7$, respectively or a little bit higher, but increased abruptly in Site M4 (Fig. 3). Thus, mean values of all the water quality parameters in the reach of $M4{\sim}M7$ sites were greater than any other sites. Seasonal data analysis indicated that BOD and EC in the downstream ($S3{\sim}S7$) was greater in the premonsoon than two seasons of the monsoon and postmonsoon, and no significant differences in BOD between the three seasons were found in the upstream ($S1{\sim}S2$). Empirical models of COD in the merged downriver was predicted ($R^2=0.87$, p>0.01, slope = 0.84, intercept = -1.28) well by EC. These results suggest that EC to be measured easily in the field may be used for estimations of nutrients and organic matter pollutions in the merged downriver and these linear models are cost-effective for the monitoring of the parameters.

Study of Operation Condition of Biofilter Using Fibril-form Matrix for Odor Gas Removal (악취가스 제거를 위안 섬유상 담체를 적용한 바이오필터 운전조건에 관한 연구)

  • Jeong Gwi-Taek;Lee Gwang-Yeon;Byun Ki-Young;Lee Kyoung-Min;Sunwoo Chang-Shin;Lee Woo-Tae;Park Chan-Young;Kim Do-Hyeong;Cha Jin-Myoung;Jang Young-Seon;Park Don-Hee
    • KSBB Journal
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    • v.20 no.5 s.94
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    • pp.341-344
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    • 2005
  • This research was performed for developing of biological treatment process of odor gas such as MEK, $H_2S$, and toluene, which was generated from the food waste recycling process. To establish the operational conditions of odor gas removal by small-scale biofiltration equipment, it was continuously operated by using toluene as a treating odor object. When the odor treating microorganisms were adhered to fibril form biofilter, high removal efficiency over $93\%$ was obtained by biofilm formation. At 400 ppm of inlet odor gas concentration and 10 sec of retention time, the removal efficiency was $76\%$ and $93\%$ in 1 st stage reactor and End stage reactor, respectively. However, the removal efficiency remained over $97\%$ at the operational conditions above 15 sec of retention time.

Lysophosphatidic Acid Stimulates SKOV-3 Cell Migration through the Generation of Reactive Oxygen Species via the mTORC2/Akt1/NOX Signaling Axis (리소포스타티드산은 SKOV-3 난소암세포의 mTORC2/Akt1/NOX 신호전달 기전을 통해 활성산소를 형성하고 이를 통해 세포의 이동을 촉진)

  • Eun Kyoung Kim;Seo Yeon Jin;Jung Min Ha;Sun Sik Bae
    • Journal of Life Science
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    • v.33 no.2
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    • pp.129-137
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    • 2023
  • Reactive oxygen species (ROS) play an essential role in a variety of cellular physiological phenomena. The present study assessed the signaling axis that mediates the lysophosphatidic acid (LPA)-induced migration of SKOV-3 cells. Insulin-like growth factor-1 (IGF-1) stimulated SKOV-3 cell migration in a time- and dose-dependent manner. Similarly, LPA stimulated SKOV-3 cell migration and the phosphorylation of Akt in a time- and dose-dependent manner. The pharmacological inhibition of LPA receptors (LPA1/LPA3) significantly suppressed LPA-induced SKOV-3 cell migration. However, IGF-1-induced SKOV-3 cell migration was not affected by the inhibition of LPA1 and LPA3. Pharmacological inhibition of phosphoinositide 3-kinase (PI3K) or Rho-associated kinase (ROCK) significantly suppressed LPA-induced migration, whereas the inhibition of MAPK kinase (MEK) had no effect. Inhibition of PI3K or ROCK completely suppressed LPA-induced ROS generation, and suppression of nicotinamide adenine dinucleotide phosphate oxidase (NOX) or chelation of ROS by N-acetylcysteine (NAC) blocked LPA-induced SKOV-3 cell migration. LPA-induced ROS generation was suppressed by silencing Rictor or Akt1 but not Raptor or Akt2. Silencing Rictor or Akt1 significantly suppressed LPA-induced SKOV-3 cell migration, whereas silencing Raptor or Akt2 had no effect. Finally, the overexpression of the constitutively active form Akt1 (CA-Akt1) significantly enhanced the LPA-induced migration of SKOV-3 cells. Given these results, we suggest that LPA stimulates SKOV-3 cell migration by ROS generation, which is mediated by the mTORC2/Akt1/NOX signaling axis.

Dielectric Properties of Semi-IPN Poly(phenylene oxide) Blend/$BaTiO_3$ Composites with Type of Cross-linker (가교체 종류에 따른 Semi-IPN Poly(phenylene oxide) 블렌드와 $BaTiO_3$ 복합재료의 유전특성)

  • Jang, Yong-Kyun;Lee, Ho-Il;Seong, Won-Mo;Park, Sang-Hoon;Yoon, Ho-Gyu
    • Polymer(Korea)
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    • v.33 no.3
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    • pp.224-229
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    • 2009
  • The dielectric properties of semi-IPN poly(phenylene oxide)(PPO) blend/$BaTiO_3$(BT) composites are investigated. The composites are fabricated via melt-mixing of crosslinker and peroxide in precursor PPO composite obtained by precipitating the suspension consisted of PPO, BT and toluene into methylethyl ketone, poor solvent of PPO. The permittivity of the precursor PPO composites shows higher value than that of integral-blended PPO composites by extruder and coincides with the theoretical value calculated by logarithmic rule of mixture. The blend of PPO and cross-linked triallyl isocyanurate is most effective for lowering the permittivity and loss tangent owing to the suppression of the orientation polarization of matrix. In contrast, 4,4'-(1,3-phenylene diisopropylidene) bisaniline, which has amine unit in its structure, increases the permittivity as well as loss tangent of the composite, but it has the ability to densify the matrix resin and the interfacial adhesion between the matrix and filler to improves flexural strength and modulus.

Diagnosis of Sapkyo Stream Watershed Using the Approach of Integrative Star-Plot Area (생태평가모형(Integrative Star-Plot Area)을 이용한 삽교천 수계 진단)

  • Kim, Ja-Hyun;Yeom, Dong-Hyuk;An, Kwang-Guk
    • Korean Journal of Ecology and Environment
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    • v.43 no.3
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    • pp.356-368
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    • 2010
  • In this study, we applied approach of integrative star-plot area (SPA), chemical water quality and habitat conditions (QHEI) to diagnoze ecological conditions at the eight sampling sites of Sapkyo Stream. These outcomes were compared with biological health based on the Index of Biological Integrity (IBI) using fish assemblage. And then, we evaluated the integrative ecological health condition using the star-plot method. This approach based on the sum of all the star-plot areas over these water and habitat characteristics. It was developed to reflect an integrative assessment of the ecological health in the stream. The biological health, based on the model values of IBI indicating "fair-poor" condition according to the criteria. Physical habitat health, based on the QHEI, averaged 123 indicating a "good-fair" condition. Also, chemical health, based on simply BOD values indicating "poor grade" according to the criteria of the Ministry of Environment Korea (MEK). The SPA indicating that 50% of the all was impaired condition and the most sampling sites were downstream sites influenced by the point and non-point sources. Overall our results suggest that the ecological health impact was a combined effect of eutrophication and habitat degradations in the stream. The approach of SPA can be used as a tool to evaluate the integrative health of stream environment and to identify possible causes of observed effects.

A study on the regulatory effect of p-38 MAP kinase on nitric oxide and interleukin-6 in osteoblasts (조골세포에시 p-38 MAP kinase의 nitric oxide 및 interleukin-6 생성조절에 관한 연구)

  • Lee, Kyung-Won;Lee, Doe-Hoon;Kang, Kyung-Hwa;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.33 no.3 s.98
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    • pp.199-210
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    • 2003
  • Tooth movement is the result of bone metabolism in the periodontium, where various cytokines take important roles. Interleukin-6(II-6) and nitrous oxide (NO) were reported to be secreted from osteoblasts in the process of bone resorption. The mechanism of the process has not been clearly understood, but the activation of mitogen-activated protein kinase (MAPK) was known to be an important process in the release of the inflammatory cytotines in macrophages. In this regard, to prove the role of MAPK in the release of IL-6 and NO in MC3T3E-1 osteoblasts, Northern blot analysis, Western blot analysis and immune complex kinase assay were used. As a result, the treatment of MC3T3E-1 osteoblast cultures with combined $interferon-\gamma(IFN-\gamma)$, lipopolysaccharide (LPS) and tumor necrosis $factor-\alpha(TNF-\alpha)$ induces expressions of inducible nitric oxide synthase (iNOS) and IL-6, resulting in sustained releases of large amounts of NO and IL-6. However, $IFN-\gamma,\;LPS,\;and\;TNF-\alpha$ individually induce a non-detectable or small amount of NO and IL-6 in MC3T3E-1 osteoblasts. The role of MAPK activation in the early intracellular signal transduction involved in iNOS and IL-6 transcription in the combined agents-stimulated osteoblasts has been investigated. The p38 MAPK pathway is specifically involved in the combined agents-induced NO and IL-6 release, since NO and IL-6 release in the presence of a specific inhibitor of p38 MAPK, 4-(4-fluorophenyl)-2-(4-metylsulfinylphenyl)-5-(4-metylsulfinylphenyl)-5-(4-pyridyl)imidazole) (SB203580), were significantly diminished. In contrast, PD98059, a specific inhibitor of MEK1, had no effect on NO and IL-6 release. Northern blot analysis showed that the p3a MAPK pathway controlled the iNOS and IL-6 transcription level. These data suggest that p38 MAPK play an important role in the secretion of NO and IL-6 in $LPS/IFN{\gamma}-or\;TNF-\gamma-treated\;MC3T3E-1$ osteoblasts.