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Destabilization of TNF-α mRNA by Rapamycin

  • Park, Jong-Woo;Jeon, Ye-Ji;Lee, Jae-Cheol;Ahn, So-Ra;Ha, Shin-Won;Bang, So-Young;Park, Eun-Kyung;Yi, Sang-Ah;Lee, Min-Gyu;Han, Jeung-Whan
    • Biomolecules & Therapeutics
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    • v.20 no.1
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    • pp.43-49
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    • 2012
  • Stimulation of mast cells through the high affinity IgE receptor (Fc${\varepsilon}$RI) induces degranulation, lipid mediator release, and cytokine secretion leading to allergic reactions. Although various signaling pathways have been characterized to be involved in the Fc${\varepsilon}$RI-mediated responses, little is known about the precious mechanism for the expression of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in mast cells. Here, we report that rapamycin, a specific inhibitor of mammalian target of rapamycin (mTOR), reduces the expression of TNF-${\alpha}$ in rat basophilic leukemia (RBL-2H3) cells. IgE or specific antigen stimulation of RBL-2H3 cells increases the expression of TNF-${\alpha}$ and activates various signaling molecules including S6K1, Akt and p38 MAPK. Rapamycin specifically inhibits antigeninduced TNF-${\alpha}$ mRNA level, while other kinase inhibitors have no effect on TNF-${\alpha}$ mRNA level. These data indicate that mTOR signaling pathway is the main regulation mechanism for antigen-induced TNF-${\alpha}$ expression. TNF-${\alpha}$ mRNA stability analysis using reporter construct containing TNF-${\alpha}$ adenylate/uridylate-rich elements (AREs) shows that rapamycin destabilizes TNF-${\alpha}$ mRNA via regulating the AU-rich element of TNF-${\alpha}$ mRNA. The antigen-induced activation of S6K1 is inhibited by specific kinase inhibitors including mTOR, PI3K, PKC and $Ca^{2+}$chelator inhibitor, while TNF-${\alpha}$ mRNA level is reduced only by rapamycin treatment. These data suggest that the effects of rapamycin on the expression of TNF-${\alpha}$ mRNA are not mediated by S6K1 but regulated by mTOR. Taken together, our results reveal that mTOR signaling pathway is a novel regulation mechanism for antigen-induced TNF-${\alpha}$ expression in RBL-2H3 cells.

Effect of Ethane 1,2-Dimethane Sulfonate(EDS) on the Expression of Steroid Hormone Receptors, $5{\alpha}$-reductase and Aromatase in the Rat Epididymis (흰쥐 부정소 내의 스테로이드 호르몬 수용체, $5{\alpha}$-reductase 그리고 Aromatase 발현에 미치는 EDS의 영향)

  • Son, Hyeok-Joon;Lee, Sung-Ho
    • Development and Reproduction
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    • v.11 no.3
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    • pp.187-193
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    • 2007
  • Ethane 1,2-dimethane sulfonate(EDS), a Leydig cell specific toxicant, has been widely used to create the reversible testosterone withdrawal rat model. Though the maintenance of epididymal structure and function is highly dependent on the testosterone secreted from testis, its derivatives, dihydroxytestosterone(DHT) and estrogen, might have crucial roles. The aim of present study was to monitor the expression patterns of sex steroid receptors, cytochrome P450 aromatase(P450arom) and $5{\alpha}$-reductase in the rat epididymis up to 7 weeks after EDS injection. Adult male rats($350{\sim}400g$) were injected with a single does of EDS(75 mg/kg i.p.) and sacrificed on weeks 0, 1, 2, 3, 4, 5, 6 and 7. The transcriptional activities of the target genes were evaluated by semi-quantitative RT-PCRs. The transcript level of estrogen receptor alpha($ER{\alpha}$) in EDS group was significantly higher than control level on week 1(P<0.01). After week 2, there was no significant difference in $ER{\alpha}$ levels between EDS group and control. The transcript level of estrogen receptor beta($ER{\beta}$) in EDS group was significantly higher than control level on week 1(P<0.05), lowered on weeks 2 and 3(P<0.05 and P<0.01, respectively), fluctuated during weeks 4 and 6, and elevated on week 7(P<0.05). The androgen receptor (AR) message levels increased significantly week 2(P<0.01), then returned to control level on week 3. In contrast, expression of cytochrome P450 aromatase(P450arom) decreased sharply during weeks $1{\sim}3$(P<0.01 on weeks 1 and 2; P<0.05 on week 3), then went back to control level on week 4. The mRNA level of $5{\alpha}$-reductase type 2($5{\alpha}$-RT2) increased significantly on week 4(P<0.01), then returned to control level. The present study indicated that EDS administration could induce reversible alterations in the transcriptional activities of sex steroid hormone receptors and androgenconverting enzymes in rat epididymis. EDS injection model will be useful to clarify the regulation mechanism of mammalian epididymal physiology.

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Sex Detection and In Vitro Development of Biopsied Bovine Embryo for LAMP Based Embryo Sexing (LAMP 방법에 의한 소 수정란의 성 판별과 Biopsy에 따른 수정란의 체외발달)

  • Cho S. R.;Choi S. H.;Kim H. J.;Han M. H.;Choe C. Y.;Chung Y. G.;Son D. S.
    • Journal of Embryo Transfer
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    • v.20 no.2
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    • pp.169-176
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    • 2005
  • Loop-mediated isothermal amplification (LAMP) is novel DNA amplification methods that amplifies a target sequence specifically under isothemal condition. The present study was to assess the in vitro viability afier biopsy and sexing rate of different types of embryo biopsied. In vivo compact morulae and blastocyst embryos were obtained from Korean Native Cow (KNC) superovulated with FSH (Antorin, R-10) on 7 Day after artificial insemination. in vitro compact morulae and blastocyst embryos were obtained with KNC or Holsteins that were gained on 6, 7 or 8 day after in vitro fertilization(IVF) with frozen semen. Biopsy of bovine embryo was carried out in a $80{\mu}l$ drop with $Ca^{2+}-Mg^{2+}$ free D-PBS and the viability of biopsied embryos were evaluated in IVMD (IFP, Japan) medium at 12 hrs culture time. The sex ratio of biopsied Hanwoo embryos were male vs. female of $43.5\%\;vs.\;56.5\%$ in vivo and $33.9\%\;vs.\;49.2\%$ in vitro respectively, and male rate of biopsied Holstein embryos were significantly higher than female $(70.8\;vs.\;29.2\%)$. and indefinite rate of in vitro embryos was $16.9\%$ and in vivo was not. The degeneration rate of biopsied embryo, in vitro embryos were significantly higher than in vivo $(13.2\%\;vs,\;0.0\%,\;p<0.05)$. The survivability of in vivo embryo were between biopsied following punching method was significantly (P<0.05) higher than bisection method produced embryos $(100\%\;vs.\;83.3\%)$ and in vitro had no difference. However, the degeneration rate of biopsied embryo by bisection method was significantly higher than punching methods between in vivo and in vitro $(16.7\;vs.\;22.6\%,\;respectively,\;p<0.05)$. In conclusion, these results indicate that punching method was optimal and survivability after embryo biopsy was useful for reducing the damage caused by the embryo biopsy procedure for LAMP-based embryo sexing.

Current Status and Prospects of Various Methods used for Screening Probiotic Microorganisms (Probiotic 미생물 검사에 사용되는 다양한 방법들에 대한 현황과 향후 전망)

  • Kim, Dong-Hyeon;Kim, Hong-Seok;Jeong, Dana;Chon, Jung-Whan;Kim, Hyunsook;Kim, Young-Ji;Kang, Il-Byung;Lee, Soo-Kyung;Song, Kwang-Young;Park, Jin-Hyeong;Chang, Ho-Seok;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.34 no.4
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    • pp.203-216
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    • 2016
  • Probiotic microorganisms are thought to provide health benefits when consumed. In 2001, the World Health Organization defined probiotics as "live microorganisms which confer a health benefit on the host, when administered in adequate amounts." Three methods for screening potential probiotics have currently widely available. (1) In vitro assays of potential probiotics are preferred because of their simplicity and low cost. (2) The use of in vivo approaches for exploring various potential probiotics reflects the enormous diversity in biological models with various complex mechanisms. (3) Potential probiotics have been analyzed using several genetic and omics technologies to identify gene expression or protein production patterns under various conditions. However, there is no ideal procedure for selecting potential probiotics than testing cadidate strains on the target population. Hence, in this review, we provide an overview of the different methodologies used to identify new probiotic strains. Furthermore, we describe futre perspectives for the use of in vitro, in vivo and omics in probiotic research.

The Analysis of Verbal Interaction in Elementary Science Programs Using Multi-Level Instruction (다수준 포함 교수법을 적용한 초등과학 프로그램에서의 언어적 상호작용 분석)

  • Jung, Suk-Jin;Shin, Young-Joon
    • Journal of The Korean Association For Science Education
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    • v.33 no.7
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    • pp.1450-1470
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    • 2013
  • The purpose of this study was to develop elementary science programs that used multi-level instruction and to analyze verbal interaction patterns in science classes that applied these programs. For this study, the 27 children from the fifth grade at B elementary school, located in Gyeonggi-do, were selected and separated into heterogeneous groups of four students. Verbal interactions occuring in two groups during each class were recorded using video. Elementary science programs using multi-level instruction were developed to target a fifth grade second semester 'Lesson 1. Human Body'. This program provided a mission form for each group and evaluation form for each child. A mission form depending on the children's level has different colors and levels of difficulty for questions. The evaluation form is composed of questions suitable for a child's level in reaching the goal with key concepts. The verbal interaction was mostly categorized into the cognitive domain and the affective domain for analysis. The cognitive domain was subdivided into question, response, making solution, receiving opinion, and the affective domain was divided into behavioral participation and students' attitude. Results of study showed that the frequency of the cognitive domain was higher than the frequency of the affective domain. In the cognitive domain, the median-level was of highest frequency in the children. In the affective domain, high-level was of highest frequency in the children. In terms of both the cognitive and affective domains of children, low-level exhibited the lowest frequency. Verbal interaction frequency was no difference between high-level and median level in cognitive and affective aspects, so median-level children were actively participating in activities similarly with high-level children. There were more types of interactions question, response, making solution, students' attitude in the median-low level children's verbal interaction than high-median level children's verbal interaction.

Usefulness and Comparison of $^{201}TI$-chloride, $^{99}mTc$-MIBI, $^{99}mTc(V)$-DMSA Single Photon Emission Computed Tomography in Distinguishing Lung Cancer from Benign Lesion (폐암과 양성질환의 감별에 $^{201}TI$-chloride, $^{99}mTc$-MIBI, $^{99}mTc(V)$-DMSA 단일광자전산화단층촬영술의 비교 및 가치)

  • Kim, Chang Ho;Chae, Sang Cheol;Park, Jae Yong;Jung, Tae Hoon;Ahn, Byeong Cheol;Lee, Jae Tae
    • Tuberculosis and Respiratory Diseases
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    • v.43 no.5
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    • pp.720-727
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    • 1996
  • Objectives: $^{201}TI$ - chloride, $^{99m}Tc$ - MIBI, $^{99m}Tc$(V) - DMSA SPECT has been used in distinguishing lung cancer from benign lesion. To compare the diagnostic efficacy of SPECT with these tumor - seeking agents, we perfonned three consecutive SPECT using $^{201}TI$, $^{99m}Tc$ - MIBI, $^{99m}Tc$(V) - DMSA in same subjects with a solitary pulmonary lesion. Methods: SPECT was carried out at 10min and 3hr for $^{201}TI$ after injection of 20mCi, and 2hr for $^{99m}Tc$ - MIBI and $^{99m}Tc$(V) - DMSA after injection of 20mCi, respectively, in 37 patients with a solitary pulmonary lesion(27 lung cancer and 10 benign diseases). In patients showing visual uptake on lesion site, we obtained the lesion - to - background(target lesion/contralateral normal lung) uptake ratio from transverse slice for each radionuclide and also calculated the retention index for $^{201}TI$. Results: The diagnostic sensitivity of $^{201}TI$, $^{99m}Tc$ - MIBI and $^{99m}Tc$(V) - DMSA SPECT to lung cancer was 100%, 96% and 73%, and the specificity was 40%, 70% and 70%, respectively. The low specificities for these agents were mainly due to high positive uptake in patients with active pulmonary tuberculosis. There were no significant differences in uptake ratios and retention index between malignant and benign lesions, and among the histologic types of lung cancer. Conclusion : $^{201}TI$ and $^{99m}Tc$ - MIBI showed higher sensitivity than $^{99m}Tc$(V) - DMSA for detecting lung cancer, but was of limited usefulness in distinguishing lung cancer from benign lesion due to low specificity, especially in area with a high prevalence of active pulmonary tuberculosis.

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Routine Off-pump Total Arterial Coronary Revascularization (심폐바이때스 없이 시행된 동맥 도관만를 이용한 관상동맥 완전 재관혈화)

  • Lee, Jae-Won;Park, Nam-Hee;Kang, Seong-Sik;Choo, Suk-Jung;Park, Seung-Jung;Park, Seung-Wook;Hong, Myeong-Ki;Song, Hyun;Song, Meong-Gun
    • Journal of Chest Surgery
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    • v.36 no.5
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    • pp.309-315
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    • 2003
  • Background: To avoid the adverse effects of cardiopulmonary bypass and to overcome late vein graft failure we routinely peformed off-pump total arterial coronary revascularization. Material and Method: From July 2000 to August 2001, 104 consecutive patients underwent first elective off-pump total arterial coronary revascularization. Both internal mammary, radial and gastroepiploic arteries were used. Sequential and composite grafts were used to achieve complete revascularization. Perioperative adverse events and postoperative angiograms were analyzed. Result: A total of 252 arterial conduits were used with an average of 2.47 grafts per patient. A total of 326 distal anastomosis were performed with a mean of 3.13 distal anastomosis per patient. Cross over to on-pump occurred in seven patients (6.7%). Of these 4 were due to unstable hemodynamics during lateral or posterior wall stabilization as a result of cardiomegaly and 3 were due to uncontrolled bleeding during dissection of diffusely dimunitive deeply placed intramyocardial coronary arteries. There were no opeartive deaths. Two cases of perioperative myocardial infarction and transient neurologic complications occurred, respectively. Of the 312 distal anastomoses, 308 (98.7%) were compatible with Fitz-Gibboll A or B patency grading. Conclusion: Off-pump total arterial coronary revascularization was technically feasible in most elective cases with satisfactory early results. However, on-pump coronary bypass surgery should be considered in difficult circumstances, such as cardiomegaly or unfavorable anatomy of the target coronary artery.

Preliminary Results of Stereotactic Radiosurgery Using Stereotactic Body Frame (정위 체부 고정틀을 이용한 체부 방사선수술의 예비적 결과)

  • Ahn Seung Do;Yi Byong Yong;Choi Eun Kyung;Kim Jong Hoo;Nho Young Ju;Shin Kyung Hwan;Kim Kyoung Ju;Chung Won Kyun;Chang Hyesook
    • Radiation Oncology Journal
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    • v.18 no.4
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    • pp.251-256
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    • 2000
  • Purpose : To evaluate efficacy and complication of stereotactic radiosurgery using stereotactic body frame. Methods and Materials :From December 1997 to June 1999, 11 patients with primary and metastatic tumors were treated with stereotactic radiosurgery using stereotactic body frame(Precision TherapyTu). Three patients were treated with primary hepatoma and seven with metastatic tumor from liver, lung, breast, trachea and one with arteriovenous malformation on neck. We used vacuum pillow for immobilization and made skin marker on sternum and tibia area with chest marker and leg marker. Diaphragm control was used for reducing movement by respiration. CT-simulation and treatment planning were peformed. Set-up error was checked by CT-Simulator before each treatment. Dose were calculated on the 80$\~$90$\%$ isodose of isocenter dose and given consecutive 3 fractions for total dose of 30 Gy (10 Gy/fraction). Results :Median follow-up was 12 months. One patient (9$\%$) showed complete response and four Patients (36$\%$) showed partial response and others showed stable disease. Planning target volumes (PTV) ranged from 3 to 111 cc (mean 18.4 n). Set-up error was within 5 mm in all directions (X, Y, Z axis). There was no complication in all patients. Conclusion :In Primary and metastatic tumors, stereotactic body frame is very safe, accurate and effective treatment modality.

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A study to 3D dose measurement and evaluation for Respiratory Motion in Lung Cancer Stereotactic Body Radiotherapy Treatment (폐암의 정위적체부방사선치료시 호흡 움직임에 따른 3D 선량 측정평가)

  • Choi, Byeong-Geol;Choi, Chang-Heon;Yun, Il-Gyu;Yang, Jin-Seong;Lee, Dong-Myeong;Park, Ju-Mi
    • The Journal of Korean Society for Radiation Therapy
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    • v.26 no.1
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    • pp.59-67
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    • 2014
  • Purpose : This study aims to evaluate 3D dosimetric impact for MIP image and each phase image in stereotactic body radiotherapy (SBRT) for lung cancer using volumetric modulated arc therapy (VMAT). Materials and Methods : For each of 5 patients with non-small-cell pulmonary tumors, a respiration-correlated four-dimensional computed tomography (4DCT) study was performed. We obtain ten 3D CT images corresponding to phases of a breathing cycle. Treatment plans were generated using MIP CT image and each phases 3D CT. We performed the dose verification of the TPS with use of the Ion chamber and COMPASS. The dose distribution that were 3D reconstructed using MIP CT image compared with dose distribution on the corresponding phase of the 4D CT data. Results : Gamma evaluation was performed to evaluate the accuracy of dose delivery for MIP CT data and 4D CT data of 5 patients. The average percentage of points passing the gamma criteria of 2 mm/2% about 99%. The average Homogeneity Index difference between MIP and each 3D data of patient dose was 0.03~0.04. The average difference between PTV maximum dose was 3.30 cGy, The average different Spinal Coad dose was 3.30 cGy, The average of difference with $V_{20}$, $V_{10}$, $V_5$ of Lung was -0.04%~2.32%. The average Homogeneity Index difference between MIP and each phase 3d data of all patient was -0.03~0.03. The average PTV maximum dose difference was minimum for 10% phase and maximum for 70% phase. The average Spain cord maximum dose difference was minimum for 0% phase and maximum for 50% phase. The average difference of $V_{20}$, $V_{10}$, $V_5$ of Lung show bo certain trend. Conclusion : There is no tendency of dose difference between MIP with 3D CT data of each phase. But there are appreciable difference for specific phase. It is need to study about patient group which has similar tumor location and breathing motion. Then we compare with dose distribution for each phase 3D image data or MIP image data. we will determine appropriate image data for treatment plan.

The Effect of Subject Well-being on the Consumer's Pricing of Alternatives (주관적 행복이 대안에 대한 소비자의 가격 책정에 미치는 영향)

  • Kim, Moon-Seop;Choi, Jong-An
    • Journal of Distribution Science
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    • v.10 no.4
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    • pp.29-36
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    • 2012
  • Research on subjective well-being (SWB) has flourished in recent years. As SWB determines cognitive and motivational processes, including social comparison and cognitive dissonance, it determines how consumers make decisions, including the comparison and evaluation of alternatives. Considering that the comparison and evaluation of alternatives is related to social comparison and cognitive dissonance, the influence of SWB on the comparison and evaluation of alternatives needs to be investigated. This research aims to examine the effect of SWB on the comparison and evaluation of alternatives, especially when people acquire additional information about their chosen or non-chosen alternatives, leading to a change of absolute/relative value of alternatives. The reasonable price of an alternative as evaluated by individuals is used as a measure reflecting the perceived value of an alternative. Putting all of this together, the current study intended to investigate the influence of absolute and relative value on the reasonable price of an alternative depending on SWB. Participants were randomly assigned to one of two experiment groups (deterioration of non-chosen alternative vs. improvement of non-chosen alternative). After reading consumer report ratings of alternatives shown on monitor screens, participants chose one of the alternatives, followed by the change of the consumer report ratings (deterioration of non-chosen alternative vs. improvement of non-chosen alternative). Participants evaluated the reasonable price of their chosen alternative based on the provided price of the non-chosen alternative. Two weeks after the experiment, they were asked to answer survey questionnaire on SWB measures. A regression was performed on the reasonable price with experiment groups, mean-centered SWB, and their interaction. There was a significant simple effect of groups and SWB. More importantly, these effects were qualified by the predicted interaction of groups and SWB. To interpret this interaction further, simple slope tests were performed on the price when SWB was centered at one standard deviation above (i.e., happy people) and below (i.e., unhappy people) the mean. As predicted, happy people rated the reasonable price of the chosen alternative higher in the improvement of non-chosen alternative group than in the deterioration of non-chosen alternative group. Conversely, unhappy people showed no price difference between groups. These results show that happy people pay attention to the absolute value of the alternative, whereas unhappy people give more weight to the relative value as well as to the absolute value of a chosen alternative, indicating that unhappy people are more sensitive to the negative information of a non-chosen alternative compared to happy people. The present research expanded the existing research stream on SWB by showing the influence of SWB on the consumers' evaluation of alternatives. Furthermore, this study adds to previous research on SWB and social comparison by suggesting that unhappy people tend to be more sensitive to negative social comparison information of alternatives even when a target of social comparison is not explicitly present. Moreover, these results yield some managerial implications on how to provide product information based on SWB in order to make products more attractive among the alternatives available to consumers.

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