• Title/Summary/Keyword: Activators

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Identification of HUGT1 as a Potential BiP Activator and a Cellular Target for Improvement of Recombinant Protein Production Using a cDNA Screening System

  • Ku, Sebastian Chih Yuan;Lwa, Teng Rhui;Giam, Maybelline;Yap, Miranda Gek Sim;Chao, Sheng-Hao
    • Molecules and Cells
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    • v.27 no.5
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    • pp.577-582
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    • 2009
  • The development of a high-throughput functional genomic screening provides a novel and expeditious approach in identifying critical genes involved in specific biological processes. Here we describe a cell-based cDNA screening system to identify the transcription activators of BiP, an endoplasmic reticulum (ER) chaperone protein. BiP promoter contains the ER stress element which is commonly present in the genes involved in unfolded protein response (UPR) that regulates protein secretion in cells. Therefore, the positive regulators of BiP may also be utilized to improve the recombinant protein production through modulation of UPR. Four BiP activators, including human UDP-glucose:glycoprotein glucosyltransferase 1 (HUGT1), are identified by the cDNA screening. Overexpression of HUGT1 leads to a significant increase in the production of recombinant erythropoietin, interferon ${\gamma}$, and monoclonal antibody in HEK293 cells. Our results demonstrate that the cDNA screening for BiP activators may be effective to identify the novel BiP regulators and HUGT1 may serve as an ideal target gene for improving the recombinant protein production in mammalian cells.

Plasminogen Activators Activities in Oviductal Epithelial Cells during Estrus Cycle in the Pig

  • Shin, Mi-Young;Kim, Tae-Shin;Kwon, Eun-Hye;Park, Soo-Bong;Park, Chun-Keun;Lee, Dong-Seok
    • Reproductive and Developmental Biology
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    • v.32 no.2
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    • pp.89-95
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    • 2008
  • The present study was undertaken to identify changes of plasminogen activators (PAs) in porcine oviductal epithelial cells (POECs) during the estrous cycle classified with post-ovulatory stages (Post-Ov), early to mid-luteal stages (Early-mid L) and pre-ovulatory (Pre-Ov) stages. The urokinase-type plasminogen activator (uPA) was only observed on day 5 and day 7 of culture in the POECs on all the estrous cycles and gradually increased according to increasing culture times, but not Early-mid L. In POECs-conditioned medium, uPA, tissue-type (tPA) and tPA-PA inhibitor (tPA-PAI) activity were observed at all culture times during estrous cycles. The uPA activity of POECs-conditioned medium on Post-Ov stage were significantly (p<0.05) decreased during prolonged cultures. On the other hand, the tPA activity of POECs-conditioned medium at Post-Ov stage was significantly (p<0.05) higher on day 5 than compared to the other days. Although was higher on day 1 at Post-Ov stage, the tPA-PAI activity of POECs-conditioned medium was significantly (p<0.05) higher on day 7 at all stage than that of day 5 of the culture. Taken together, these results suggest that uPA, tPA and tPA-PAI are produced by POECs, and the variations of the PAs activity are regulated in the different stages of the estrous cycle.

Analysis of Plasminogen Activators Activity and Three Dimensional (3D) Culture of Endometrial Cells in Pigs (돼지 자궁내막 세포의 3차원 배양과 Plasminogen Activator 활성화 분석)

  • Cha, Hye-Jin;Lee, Sang-Hee;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Journal of Embryo Transfer
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    • v.28 no.3
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    • pp.273-280
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    • 2013
  • The aim of this study was to establish a three dimensional (3D) culture system of endometrial cells and to examine the plasminogen activators (PAs) activity in porcine uterine. The 3D culture system in porcine endometrial cells was composed to mixture 3D gel, stromal cells and epithelial cells. The 3D culture system was used to identify normal structure search as uterine tissue and PAs expression in this study. In results, porcine endometrium epithelial cells forming a top monolayer and endometrium stromal cells developed as fibroblast-like within 3D matrix scaffold. Expression of urokinase-type PA (uPA) and tissue-type PA (tPA) were observed during the 3D culture using immunofluorescence. PA activity in 3D-cultured endometrial cells was no significant difference between the tissue type, but 2D culture system were significantly lower than in 3D-cultured endometrial cells (P<0.05). Therefore, basic system and functional aspect of 3D culture could be established with similar system of endometrium tissue. We suggest that this study was assumed applicable as baseline data to investigate mechanism between porcine uterus cells in vitro.

Effect of two root activators [phthalamic acid and N(p - chlorophenyl) phthalamic acid] on Rice yield (근활력(根活力) 증진제(增進劑) [phthalamic acid 와 N(p - chlorophenyl) phthalamic acid] 의 수도증수(水稻增收) 효과(效果))

  • Park, Hoon;Mok, Sung-Kyun
    • Applied Biological Chemistry
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    • v.17 no.4
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    • pp.235-239
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    • 1974
  • Rice root activiators (phthalamic acid (PA) and p-chlorophenyl PA (PCPPA) were tested in pot culture for their effects on yield of IR667-Suwon 214 weak in root activity. The results were as follows: 1) Maximum percentage yield increase was 36.7% with organic matter applied and 13.3% without organic matter in 100 ppm (90ml/0.05$m^2$) of PCPPA treatment. 2) Root activators increased oxidizing power of root (root activity), consequently gave a balanced nutrient uptake, stimulated root growth and subsequent shoot growth. 3) Root activators increased number of panicle per hill, grain weight, filled grain ratio and effective tiller ratio.

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Action of Protein Kinase A and C Activators on Germinal Vesicle Breakdown and One-Cell Embryos in the Mouse (생쥐 GV난자와 1-세포기 배아의 핵막붕괴에 미치는 Protein Kinase A와 C의 작용)

  • 이대기;김경진;조완규
    • The Korean Journal of Zoology
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    • v.32 no.2
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    • pp.153-162
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    • 1989
  • Expedments were perfonned to examine the role of cAMP-dependent protein kinase (PK-A) and diacylglycerol-dependent protein kinase (PK-C) during the meiodc resumption and the first mitotic cell cycle of mouse embryogenesis. Mejoric GV oocytes and one-cell embryos derived from in vitro fertilization were cultured in vitro, and morphological changes in response to activators of PK-A and PK-C were examined. Treatments with a membrane-permeable cAMP analog, dbcAMP (0.1 mg/mi), phosphodiesterase inhibitor, IBMX (0.1 mM), biologically active phorbol ester, WA (10 nglmi), or a synthetic diacylglycerol, sn-diC8 inhibited resumption of melosis. Combination of PK-A and PK-C activator brought about furiher inhibition. On the contrary, dbcAMP (0.1 mg/mi), IBMX (0.2 mM), WA (10 nglml), and sn-diC8 (0.5 mM) did not inhibit pronucleus membrane breakdown (PNBD) when added S or G2 phase of cell cycle. However, activators of PK-C inhibited cleavage of one-cefl embryos. This result indicates that the action mechanism of PK-A and PK-C on dissolution of nuclear membrane in primary meiotic arrest oocytes may be different from that of mitotic one-cell embryos.

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Effects of Progesterone and 17β-Estradiol under Presence or Absence of FBS on Plasminogen Activators Activity in Porcine Uterine Epithelial Cells

  • Hwangbo, Yong;Lee, Mi-Rim;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Development and Reproduction
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    • v.22 no.4
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    • pp.309-318
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    • 2018
  • The present study was conducted to investigate the regulatory mechanism of plasminogen activators (PAs) activation by $17{\beta}$-estradiol ($E_2$) and progesterone ($P_4$) in porcine uterine epithelial cells (pUECs). pUECs were collected from porcine uterine horn and cultured at 80% confluence. Then, 0.1% (v/v) DMSO, 20 ng/mL $E_2$, and $P_4$ with or without fetal bovine serum (FBS) treated to cultured cells for 24 hours. The supernatants were used for measurement of PAs activity and expression of urokinase-type PA (uPA), tissue-type PA (tPA), uPA specific receptor (uPAR), and type-1 PA inhibitor (PAI-1) mRNA were analyzed by real-time PCR. The expression of PAs-related genes was not affect by steroid hormones in both of serum treatment groups. However, PAs activity was increased by treatment of $E_2$ compared to 0.1% DMSO treatment in serum-free group (p<0.05). Then, $E_2$ and $P_4$ were diluted with 0.002% (v/v) DMSO for reduction of its effect and treated to cultured cells without FBS. Only tPA mRNA was significantly increased by $E_2$ treatment (p<0.05). PAs activity was enhanced in $E_2$ treated group compared to control groups (p<0.05). These results indicate that serum-free condition is more proper to evaluate effect of steroid hormones and activation of PAs in pUECs was mainly regulated by estrogen. These regulation of PAs activation may be associated with uterine remodeling during pre-ovulatory phase in pigs, however, further studies are needed to investigate precise regulatory mechanism.

The Estimation of Optimal Mixing Ratio of CLSM Mixed with Red Mud and Paper Sludge Ash (Red mud와 제지회를 혼합한 CLSM의 적정 혼합비 산정)

  • Roh, Seongoh;Kim, Taeyeon;Lee, Bongjik
    • Journal of the Korean GEO-environmental Society
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    • v.23 no.4
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    • pp.21-27
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    • 2022
  • Recently, numerous studies are being performed to examine alkali-activated cement which uses industrial by-products, such as GGBS and fly ash, as well as alkali activators. Alkali-activated cement is a type of binder that exerts the same strength as cement without using cement by mixing industrial by-products with alkali activators. Alkali activators, which are used mainly for carbon-reducing technologies and alkali activation, are expensive and difficult to apply in the field due to risks related to strong alkalinity. Therefore, this study intends to explore methods to use red mud as a substitute for an alkali activator. To that end, this study has evaluated engineering properties, such as flow and strength, of CLSM that uses red mud and paper sludge ash as binders and its possibility to cause soil pollution. This study also aims to present the appropriate mixing ratios of red mud and paper sludge ash to produce CLSM.

Effects of activators contained in adhesives on dentin bond strengths (상아질 접착제에 포함된 활성제의 사용 유무가 자가중합 복합레진의 상아질에 대한 전단결합강도에 미치는 영향)

  • Kang, Hye-Kyung;Shin, Joo-Hee;Suh, Kyu-Won;Ryu, Jae-Jun
    • The Journal of Korean Academy of Prosthodontics
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    • v.46 no.5
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    • pp.511-519
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    • 2008
  • Statement of problem: An incompatibility between the initiator systems of self-curing composite resins and light-curing adhesives was supposed recently. Purpose: The purpose of the study was to evaluate the influence of activators for self/dual bonding on dentin shear bonding strengths. Material and methods: Fifty human molars were divided into 5 groups. A flat dentin surface was created for each tooth. A self-curing composite resin (Luxacore) was bonded with the following adhesives (n = 10); One-Step, Prime&Bond NT, AdheSE, Prime&Bond NT and AdheSE were also used in combination with activators. Shear bond strengths were measured after 24 hours of water storage. The specimens were loaded in shear in the Instron until failure at a 1 mm/min crosshead speed. Data were compared using one-way ANOVA and Tukey HSD test (${\alpha}$= 0.05). Results: The dentin adhesive systems in order of decreasing median bond strength were One-Step > Prime&Bond NT with activator, AdheSE with activator > Prime&Bond NT, AdheSE. Among adhesives, One-Step had the highest bond strength. Prime&Bond NT with activator had higher bond strengths than Prime&Bond NT that was used alone, and so was AdheSE. Conclusion: Shear bond strengths were increased in Prime&Bond NT and AdheSE when these were used with activators comparing used without activators. But using activators was not effective clinically comparing One-Step.