• Title/Summary/Keyword: hRA

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Optimization of Human Embryonic Stem Cells into Differentiation of Dopaminergic Neurons in Vitro: II. Genetically Modified Human Embryonic Stem Cells Treated with RA/AA or b-FGF

  • 신현아;김은영;이영재;이금실;조황윤;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.75-75
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    • 2003
  • Since the establishment of embryonic stem cell, pluripotency of the cells was known to allow differentiation of the cells into various cell types consisting whole body. Several protocols have been developed to induce expression of specific genes.. However, no precise protocol that will generate a single type of the cells from stem cells has been reported. In order to produce cells suitable for transplantion into brain of PD animal model, which arouse due to a progressive degeneration of dopaminergic neurons in midbrain, human embryonic stem cell (hESC, MB03) was transfected with cDNAs cording for tyrosine hydroxylase (TH). Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by the two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA/ascorbic acid (AA), embryoid bodies (EB, for 4days) derived from hES cells were exposed to RA (10$^{-6}$ M)/AA (50 mM) for 4 days, and were allowed to differentiate in N2 medium for 7, 14, 21, or 28 days. Exp. II) When bFGF was used, neuronal precursor cells were selected for 8 days in N2 medium after EB formation. After selection, cells were expanded at the presence of bFGF (20 ng/ml) for another 6 days followed by a final differentiation in N2 medium for 7, 14, 21 or 28 days. By indirect immunocytochemical studies, proportion of cells expressing NF200 increased rapidly from 20% at 7 days to 70 % at 28 days in RA/AA-treated group, while those cells expressing NF160 decreased from 80% at 7 days to 10% at 28 days upon differentiation in N2 medium. However, in differentiation by RA/AA treatment system, there was a significant increase in proportion of neuron maturity (73%) at day 14 after N2 medium. TH#2/MB03 cells expressing TH are >90% when matured at the absence of either bDNF or TGF-$\alpha$. These results suggested that TH#2/MB03 cells could be differentiated in vitro into mature neurons by RA/AA.

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Comparison of Histamine 2 Receptor Antagonists and Proton Pump Inhibitors on Infectious Complications in Critically Ill Patients (중환자에서 스트레스성 궤양 예방 약물에 따른 감염성 합병증 발생률 비교)

  • Park, Sun young;Choi, Jae Hee;Youn, Young Ju;Rhie, Sandy Jeong
    • Korean Journal of Clinical Pharmacy
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    • v.26 no.1
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    • pp.46-52
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    • 2016
  • Background: The use of acid suppressive agents became a standard therapy in an intensive care unit (ICU) to prevent stress related gastrointestinal mucosal damage. However, the risk of infectious diseases has been concerned. Objective: The study was to determine the differences between histamine 2 receptor antagonists (H2RA) and proton pump inhibitors (PPI) in incidence of nosocomial pneumonia and pseudomembranous colitis (PMC) by Clostridium difficile with patients in ICU. Methods: This is a retrospective comparative study including patients admitted to the ICU who were at least 18 years of age and stayed for more than 48hrs from August 1, 2014 to January 31, 2015. The propensity score analysis and propensity matched multivariable logistic regression were used in analyzing data to control for confounders. Results: A total of 155 patients were assessed. H2RA were prescribed in 110 (53.9%) and PPI were in 45 (22.1%). Nosocomial pneumonia developed in 37 (23.9%); 25 (22.7%) were on H2RA and 12 (26.7%) were on PPI. The unadjusted incidence of nosocomial pneumonia was slightly higher in the patients with PPI (odds ratio (OR) 1.24; 95% confidence interval (CI): 0.54-2.71) compared to them with H2A. After adjusting with propensity score, the adjusted OR with PPI was 1.35 (95% CI: 0.44-4.11). The propensity score matched analyses showed similar results. Conclusion: The uses of PPI and H2RA as a stress ulcer prophylaxis agent showed similarity in the incidence of nosocomial pneumonia and PMC.

Design of a Greenhouse Monitoring System using Arduino and Wireless Communication (아두이노와 무선통신을 이용한 온실 환경 계측 시스템 설계)

  • Sung, Bo Hyun;Cho, Young-Yeol
    • Journal of Bio-Environment Control
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    • v.31 no.4
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    • pp.452-459
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    • 2022
  • One of the important factors among the smart farm factors is environmental measurement. This study tried to design an environmental measurement monitoring system through Bluetooth wireless communication with LoRa using the open source programs Arduino, App Inventor, and Node Red. This system consists of Arduino, LoRa shield, temperature and humidity sensor (SHT10), and carbon dioxide sensor (K30). The environmental measurement system is configured as a system that allows the sensor to collect environmental data and transmit it to the user through wireless communication to conveniently monitor the farm environment. As libraries used in the Arduino program, LoRa.h, Sensirion.h, LiquidCrystal_I2C.h and K30_I2C.h were used. When receiving environmental data from the sensor at regular intervals, coding using average value was used for data stabilization. An Android-based app was developed using Node Red and App Inventor program as the user interface. It can be seen that the environmental data for the sensor is well collected with the screen output to the serial screen of Arduino, the screen of the smartphone, and the user interface of Node Red. Through these open source-based platforms and programs will be applied to various agricultural applications.

Suppressive Effects of GST on Cytokine-induced Activation of Human Fibroblast-like Sinoviocytes (가미사물탕(GST)의 사이토카인으로 유도된 인간 섬유아세포양 활막 세포 활성화 저해 작용)

  • Park, Jee-Young;Jin, Mi-Rim;Kim, Dong-Hee
    • Journal of Haehwa Medicine
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    • v.14 no.2
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    • pp.45-54
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    • 2005
  • GST, an extract from 16 herbs, has been formulated and prescribed for the treatment of human rheumatoid arthritis(hRA) for many years. The present study was done to investigate whether GST has suppressive effects on activation of fibroblast-like sinoviocytes isolated from an RA patient. In tumor necrosis factor-a(TNF-a)/interleukin-1b(IL-1b) treated human sinoviocytes, The mRNA expression of molecular indicators related to pathologic changes of the sinoviocytes were examined using quantitative real-time PCR. The treatment of GST($100\;{\mu}g/ml$) suppressed the expression of proinflammatory cytokines and chemokines such as TNF-a, IL-1b, IL-6 and IL-8 compared with the control. The mRNA level of intracellular adhesion molecule-1(ICAM-1) which is known to increase in the activated sinoviocytes of RA patients, was slightly decreased by GST. The expression of NOS-II was considerably reduced, which was accompanied by a decrease in the production of nitric oxide(NO). In addition, GST considerably increased the mRNA levels of tissue inhibitors of matrix metalloproteinase-1(TIMP-1), while those of matrix metalloproteinase-3(MMP-3) were decreased. Taken together, these data suggested that GST might suppress the activation of sinoviocytes in hRA.

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Inhibitory Effects of GC, an Extract from Herbs, on $TNF-{\alpha}/IL-1{\beta}$-induced Activation of Human Fibroblast-like Sinoviocytes (계혈등복합방(GC)의 $TNF-\alpha$$IL-1{\beta}$로 유도된 인간 섬유아세포양 활막 세포 활성화 억제 작용)

  • Jang Kwang-Ho;Jin Mi-Rim;Park Hee-Ok;Kim Dong-Hee
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.5
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    • pp.1225-1232
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    • 2005
  • Based on traditional medicine theories, GC, an extract from 5 herbs, has been formulated and prescribed for the treatment of human rheumatoid arthritis(hRA) for many years. The present studies was done to investigate whether GC has inhibitory effects on activation of fibroblast-like sinoviocytes isolated from a RA patient. In tumor necrosis factor-${\alpha}(TNF-{\alpha}$)/ interleukin-IL-$1{\beta}$(IL-$1{\beta}$) treated human sinoviocytes, the mRNA expression of molecular indicators related to pathologic changes of the sinoviocytes were examined using quantitative real-time PCR. The treatment of GC($10{\mu}g/ml$) significantly suppressed the expression of proinflammatory cytokines and chemokines such as $TNF-{\alpha}$, IL-6 and IL-8 compared with the control, but not $IL-1{\beta}$, The mRNA level of intracellular adhesion molecule-1 (ICAM-1) which is known to increase in the activated sinoviocytes of RA patients, was slightly decreased by GC. The expression of NOS-II was considerably reduced, which was accompanied by a decrease in the production of nitric oxide(NO). Furthermore, GC dramatically raised the mRNA levels of tissue inhibitors of matrix metalloproteinase-1 (TIMP-1), while those of matrix metalloproteinase-3 were significantly lowered. Taken together, these data suggested that GC might suppress the activation of sinoviocytes in hRA.

In vitro Neural Cell Differentiation of Genetically Modified Human Embryonic Stem Cells Expressing Tyrosine Hydroxylase (Tyrosine Hydroxylase 유전자가 주입된 인간 배아줄기세포의 체외 신경세포 분화)

  • Shin, Hyun-Ah;Kim, Eun-Young;Lee, Keum-Sil;Cho, Hwang-Yoon;Kim, Yong-Sik;Lee, Won-Don;Park, Se-Pill;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.31 no.1
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    • pp.67-74
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    • 2004
  • Objective: This study was to examine in vitro neural cell differentiation pattern of the genetically modified human embryonic stem cells expressing tyrosine hydroxylase (TH). Materials and Methods: Human embryonic stem (hES, MB03) cell was transfected with cDNAs cording for TH. Successful transfection was confirmed by western immunoblotting. Newly transfected cell line (TH#2/MB03) was induced to differentiate by two neurogenic factors retinoic acid (RA) and b-FGF. Exp. I) Upon differentiation using RA, embryoid bodies (EB, for 4 days) derived from TH#2/MB03 cells were exposed to RA ($10^{-6}M$)/AA ($5{\times}10^{-2}mM$) for 4 days, and were allowed to differentiate in N2 medium for 7, 14 or 21 days. Exp. II) When b-FGF was used, neuronal precursor cells were expanded at the presence of b-FGF (10 ng/ml) for 6 days followed by a final differentiation in N2 medium for 7, 14 or 21 days. Neuron differentiation was examined by indirect immunocytochemistry using neuron markers (NF160 & NF200). Results: After 7 days in N2 medium, approximately 80% and 20% of the RA or b-FGF induced Th#2/MB03 cells were immunoreactive to anti-NF160 and anti-NF200 antibodies, respectively. As differentiation continued, NF200 in RA treated cells significantly increased to 73.0% on 14 days compared to that in b-FGF treated cells (53.0%, p<0.05), while the proportion of cells expressing NF160 was similarly decreased between two groups. However, throughout the differentiation, expression of TH was maintained ($\sim$90%). HPLC analyses indicated the increased levels of L-DOPA in RA treated genetically modified hES cells with longer differentiation time. Conclusion: These results suggested that a genetically modified hES cells (TH#2/MB03) could be efficiently differentiated in vitro into mature neurons by RA induction method.

Obtaining the zwitterionic form of L-lysine from L-lysine monohydrochloride by electrodialysis

  • Aghajanyan, A.E.;Tsaturyan, A.O.;Hambardzumyan, A.A.;Saghyan, A.S.
    • Membrane and Water Treatment
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    • v.4 no.1
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    • pp.1-9
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    • 2013
  • The process of electromembrane transformation of L-lysine monohydrochlorides into their zwitterionic form in four- and two-chamber electrodialysis apparatus was investigated. The process of transformation at various concentrations of lysine monohydrochloride (0.1-0.6 mol.L-1) was studied and it was established that at the optimum density of current optimal concentrations of lysine hydrochloride during electrodyalisis was in the range of 0.2-0.4 mol.L-1. It was determined that the process of total transformation was accomplished when pH of the lysine solution achieved 10. Changes of concentrations of $Cl^-$ ions and lysine diffused into the neighboring chamber were determined depending on the time. The method developed by us allows adjusting the removal coefficient of $Cl^-$ ions during transformation to a maximal value, the losses of lysine diffused into the next chamber after its return to the technological cycle being less than 1.0 %. The specific energy consumption during the process of transformation in two- and four-chamber electrodialyzers was 0.19 and 0.205 A.h.kg-1 and the current efficiency was 75.9 and 73.1 %, correspondingly. Study of the process of electromembrane transformation allowed obtaining zwitterionic form of L-lysine from L-lysine monohydrochloride with minimal reagent and energy consumption.

Effect of Retinoic Acid and dibutyryl cyclic AMP on G1 Phase Associated Molecules during F9 Embryonic Carcinoma Cell Differentiation (Retinoic acid와 dibutyryl cyclic AMP가 F9 embryonic carcinoma cell 분화 중 G1 Phase 관련 분자에 미치는 영향)

  • 박귀례;김건홍;한순영;이유미;장성재
    • YAKHAK HOEJI
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    • v.43 no.3
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    • pp.378-384
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    • 1999
  • Retinoic acid (RA) and dibutyryl cyclic AMP (dbcAMP) induce the differentiation of the multipotent embryonic carcinoma cell line, F9 cells, into parietal endoderm like cell. The F9 cells are highly proliferative doubling approximately 12 hourse. S Phase is predominant, lasting 10 hours and G2/M phase occupies most of the remaining cycle (2 hours) and G1 phase is nearly non-existent. In this study, we showed the effect of RA and dbcAMPon the cell cycle associated molecules (especially around G1 phase) during F9 cell differentiation. Differentiation of F9 cells was induced by the combined addition of RA ($10^{-7}M$) and dbcAMP (0.5mM), and cells were harvested daily up to 4 days. Flow cytometric analysis showed the prolongation of G1 phase around 30 hours after induction. Western blot analysis revealed that the amount of cyclin D1 and cdk2 were increased at day 4. However, histone H1 kinase activity of cdk2 was decreased. These data strongly suggest that RA and dbcAMP induce the growth arrest of F9 cells at G1 phase by decreasing the activity of cdk2, although they have increased the protein contents of cyclin D1 and cdk2. The reason for the discrepancy between the H1 kinase activity and protein contents are not clear yet.

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Application of Ultrasonic Vibration Energy on Eco-superfinishing and Surface Hardening Treatment of Cold Work Roller (초음파 진동에너지를 이용한 냉간 압연롤러 표면의 환경 친화적 초정밀 사상 및 표면 경화 처리 및 시험)

  • Y.S. Pyoun;Park, J.H.;C.H. Han;Park, Y.;I.S. Cho;N. Azuma;Lee, J.H.;Kim, C.S.;Park, C.H.
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2002.05a
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    • pp.39-39
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    • 2002
  • In order to improve wear and fatigue resistance of the structural materials, especially cold work roller for 304 stainless steel, an eco-super-finishing and surface hardening treatment using ultrasonic vibration energy was developed and applied to the SKD-ll roller. The eco-super-finishing machine was designed and fabricated by DesignMecha Co, by its own technology. It was observed that the surface roughness, hardness and residual stress were changed from $Ra{\;}={\;}O.25\mu\textrm{m}$, Hv=710 and ${\sigma}$={\;}+400{\;}MPa{\;}to{\;}Ra{\;}={\;}0.165\mu\textrm{m}$, Hv = 1200 and ${\sigma}=-610$ MPa after 20 KHz micro-cold forging, which means almost equal to the 300 % improvement of life-time.

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