• Title/Summary/Keyword: knock-in

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Characterization of Oszinc626, knock-out in zinc finger RING-H2 protein gene, in Ac/Ds mutant lines of rice(Oryza sativar L.) (Zinc finger RING-H2 protein관련 Ac/Ds전이인자 삽입 변이체 Oszinc626 유전자의 특성 분석)

  • Park, Seul-Ah;Jung, Yu-Jin;Ahn, Byung-Ohg;Yun, Doh-Won;Ji, Hyeon-So;Park, Yong-Hwan;Eun, Moo-Young;Suh, Seok-Cheol;Lee, Soon-Youl;Lee, Myung-Chul
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.177-183
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    • 2008
  • Ac/Ds mutant lines of this study were transgenic rice plants, each of which harbored the maize transposable element Ds together with a GUS coding sequence under the control of a promoterless(Ds-GUS). We selected the mutants that were GUS expressed lines, because the GUS positive lines will be useful for identifying gene function in rice. One of these mutants was identified knock-out at Oszinc626(NP_001049991) gene, encoding a RING-H2 zinc-finger protein, by Ds insertion. In this mutant, while primary root development is normal, secondary root development from lateral root was very poor and seed development was incomplete compare with normal plant. RING zinc-finger proteins play important roles in the regulation of development in a variety of organisms. In the plant kingdom, a few genes encoding RING zinc-finger proteins have been documented with visible effects on plant growth and development. The consensus of the RING-H2(C3-H2-C3 type) domain for this group of protein is $Cys-X_2-Cys-X_{28}-Cys-X-His-X_2-His-X_2-Cys-X_{14}-Cys-X_2-Cys$. Oszinc626 encodes a predicted protein product of 445 amino acids residues with a molecular mass of 49 kDa, with a RING-zinc-finger motif located at the extreme end of the C-terminus. RT-PCR analysis indicated that the expression of Oszinc626 gene was induced by IAA, cold, dehydration, high-salinity and abscisic acid, but not by 2,4-D, and the transcription of Oszinc626 gene accumulated primarily in rice immature seeds, root meristem and shoots. The gene accumulation patterns were corresponded with GUS expression.

A Study on the Stiffness of CBA(Corner Block with Anchor Bolt) Joint in Knockdown Type Table Furniture (조립식(組立式) 탁자(卓子)의 CBA접합부(接合部) 강성(剛性)에 관(關)한 연구(硏究))

  • Chung, Woo-Yang;Lee, Phil-Woo
    • Journal of the Korean Wood Science and Technology
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    • v.17 no.2
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    • pp.34-64
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    • 1989
  • Corner block with anchor bolt(CBA) joint method used in knock-down type table furniture manufacturing can reduce the packing and transporting cost. Unfortunately. it also has the disastrous defect to be loosend and unstable during the service life mainly due to fatigue and creep(repeated and prolonged loading). So 22 joint groups constructed were tested to evaluate the effect of some design factors related to the size of side rail(apron). block attachment to side rail. and the number of anchor bolt as well as the effect of the type of corner block(mitered type vs. rectangular type) Usable strength from the stiffness coefficients of each joint group were analysed with SPSS /PC+ and described as the criteria of CBA joint construction. The conclusions were as follows: The height of side rail(50, 75 and 100 mm) and the addition of polyvinyl acetate(PVAc) emulsion in the corner block attactment to side rail had the effect on raising the usable strength of CBA joint with remarkable high significance. And the effect of 2 - anchor bolts was also superior to that of 1 - bolt significantly. However. the thickness of side rail(22 mm vs. 25 mm) had no effect on the strengthening the table joint rigidity. Mitered type corner block joint appeared to he recommendable for CBA jointed table construction rather than the rectangular type one regardless of the method of block attachment to side rail. The best result identified from Duncan's multiple comparison was in the construction with 25 mm thick and 100 mm height of side rail fastened using 2 - anchor bolts in mitered type corner block. But it would be reasonable to use 22 mm thick & 75 mm high side rail and mitered corner block with PVAc emulsion & 2 bolts considering the productivity and production cost down in the MDF furniture manufacturing industries.

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Characterization of Cigarette Smoke Extract (CSE)-induced Cell Death in Lung Epithelial Cells (폐상피세포에서 흡연추출물-유도성 세포사에 관한 연구)

  • Choi, Eun Kyung;Kim, Yun Seup;Park, Jae Seuk;Jee, Young Koo;Lee, Kye Young
    • Tuberculosis and Respiratory Diseases
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    • v.58 no.1
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    • pp.43-53
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    • 2005
  • Emphysema is characterized by air space enlargement and alveolar destruction. The mechanism responsible for the development of emphysema was thought to be protease/antiprotease imbalance and oxidative stress. A very recent study shows that alveolar cell apoptosis causes lung destruction and emphysematous changes. Thus, this study was performed to support the evidence for the role of apoptosis in the development of emphysema by characterizing cigarette smoke extract (CSE)-induced apoptosis in A549 (type II pneumocyte) lung epithelial cells. CSE induced apoptosis at low concentration (10% or less) and both apoptosis and necrosis at high concentration (20%). Apoptosis was demonstrated by DNA fragmentation using FACScan for subG1 fraction. Discrimination between apoptosis and necrosis was done by morphologic analysis using fluorescent microscopy with Hoecst 33342/propium iodide double staing and electron microscopy. Cytochrome c release was confirmed by using immunofluorescence with monoclonal anti-cytochrome c antibody. However, CSE-induced cell death did not show the activation of caspase 3 and was not blocked by caspase inhibitors. This suggests that CSE-induced apoptosis might be caspase-independent apoptosis. CSE-induced cell death was near completely blocked by N-acetylcystein and bcl-2 overexpression protected CSE-induced cell death. This results suggests that CSE might induce apoptosis through intracellular oxidative stress. CSE also activated p53 and functional knock-out of p53 using stable overexpression of HPV-E6 protein inhibited CSE-induced cell death. The characterization of CSE-induced cell death in lung epithelial cells could support the role of lung cell apoptosis in the pathogenesis of emphysema.

Cloning and Characterization of Phosphomannomutase/Phosphoglucomutase (pmm/pgm) Gene of Vibrio anguillarum Related to Synthesis of LPS (Lipopolysaccharide 생합성에 관여하는 Vibrio anguillarum의 phosphomannomutase/phosphoglucomutase 유전자 cloning과 특성)

  • Oh, Ryunkyoung;Moon, Soo Young;Cho, Hwa Jin;Jang, Won Je;Kim, Jang-Ho;Lee, Jong Min;Kong, In-Soo
    • Microbiology and Biotechnology Letters
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    • v.44 no.3
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    • pp.355-362
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    • 2016
  • The phosphomannomutase/phosphoglucomutase gene (pmm/pgm) of Vibrio anguillarum (the causative agent of fish vibriosis) was cloned, and the open reading frame corresponded to a protein with 446 amino acids. The pmm/pgm gene showed a significant degree of sequence homology with the previously reported genes from V. mimicus, V. vulnificus, V. splendidus, and V. harveyi, with 92.3%, 91.4%, 89.9%, and 89.9% amino acid identity, respectively. By reverse transcriptase-polymerase chain reaction, we found that the pmm/pgm gene was upregulated under cold stress condition. The PMM/PGM protein is known to catalyze the interconversion between mannose-1-phosphate and mannose-6-phosphate or glucose-1-phosphate and glucose-6-phosphate, which are important intermediates for lipopolysaccharide (LPS) biosynthesis. To confirm the role of PMM/PGM in the LPS biosynthetic pathway, we constructed a knock out mutant by homologous recombination. The respective LPSs were isolated from the V. anguillarum wild-type and mutant strains, and changes were compared by subjecting them to sodium dodecyl sulfate polyacrylamide gel electrophoresis. Based on the different patterns of the LPSs, we expect the pmm/pgm gene to have an important role in LPS biosynthesis. The pmm/pgm-deficient mutant of V. anguillarum will contribute to further studies about the role of LPS in V. anguillarum pathogenesis.

A Study on Properties of Crude Oil Based Derivative Linked Security (유가 연계 파생결합증권의 특성에 대한 연구)

  • Sohn, Kyoung-Woo;Chung, Ji-Yeong
    • Asia-Pacific Journal of Business
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    • v.11 no.3
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    • pp.243-260
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    • 2020
  • Purpose - This paper aims to investigate the properties of crude oil based derivative security (DLS) focusing on step-down type for comprehensive understanding of its risk. Design/methodology/approach - Kernel estimation is conducted to figure out statistical feature of the process of oil price. We simulate oil price paths based on kernel estimation results and derive probabilities of hitting the barrier and early redemption. Findings - The amount of issuance for crude oil based DLS is relatively low when base prices are below $40 while it is high when base prices are around $60 or $100, which is not consistent with kernel estimation results showing that oil futures prices tend to revert toward $46.14 and the mean-reverting speed is faster as oil price is lower. The analysis based on simulated oil price paths reveals that probability of early redemption is below 50% for DLS with high base prices and the ratio of the probability of early redemption to the probability of hitting barrier is remarkably low compared to the case for DLS with low base prices, as the chance of early redemption is deferred. Research implications or Originality - Empirical results imply that the level of the base price is a crucial factor of the risk for DLS, thus introducing a time-varying knock-in barrier, which is similar to adjust the base price, merits consideration to enhance protection for DLS investors.

SOCS3 Attenuates Dexamethasone-Induced M2 Polarization by Down-Regulation of GILZ via ROS- and p38 MAPK-Dependent Pathways

  • Hana Jeong;Hyeyoung Yoon;Yerin Lee;Jun Tae Kim;Moses Yang;Gayoung Kim;Bom Jung;Seok Hee Park;Choong-Eun Lee
    • IMMUNE NETWORK
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    • v.22 no.4
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    • pp.33.1-33.17
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    • 2022
  • Suppressors of cytokine signaling (SOCS) have emerged as potential regulators of macrophage function. We have investigated mechanisms of SOCS3 action on type 2 macrophage (M2) differentiation induced by glucocorticoid using human monocytic cell lines and mouse bone marrow-derived macrophages. Treatment of THP1 monocytic cells with dexamethasone (Dex) induced ROS generation and M2 polarization promoting IL-10 and TGF-β production, while suppressing IL-1β, TNF-α and IL-6 production. SOCS3 over-expression reduced, whereas SOCS3 ablation enhanced IL-10 and TGF-β induction with concomitant regulation of ROS. As a mediator of M2 differentiation, glucocorticoid-induced leucine zipper (GILZ) was down-regulated by SOCS3 and up-regulated by shSOCS3. The induction of GILZ and IL-10 by Dex was dependent on ROS and p38 MAPK activity. Importantly, GILZ ablation led to the inhibition of ROS generation and anti-inflammatory cytokine induction by Dex. Moreover, GILZ knock-down negated the up-regulation of IL-10 production induced by shSOCS3 transduction. Our data suggest that SOCS3 targets ROS- and p38-dependent GILZ expression to suppress Dex-induced M2 polarization.

The Role of the Insulin-like Growth Factor System during the Periimplantation Period (착상기 Insulin-like Growth Factor System의 역할)

  • 이철영
    • Journal of Embryo Transfer
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    • v.12 no.3
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    • pp.229-246
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    • 1997
  • Implantation is a most important biological process during pregnancy whereby conceptus establishes its survival as well as maintenance of pregnancy. During the periimplantation period, both uterine endometriurn and conceptus synthesize and secrete a host of growth factors and cytokines which mediate the actions of estrogen and /or progesterone and also exert their steroid-independent actions. Growth factors expressed by the materno-conceptal unit en masse have important roles in cell migration, stimulation or inhibition of cell proliferation, cellular differentiation, maintenance of pregnancy and materno-conceptal communications in an autorcrine /paracrine manner. The present review focuses on the role of the intrauterine IGF system during periimplantation conceptus development. The IGF system comprises of IGF- I and IGF- II ligands, types I and II IGF receptors and six or more IGF-binding proteins(IGFBPs). IGFs and IGFBPs are expressed and secreted by uterine endometrium with tissue, pregnancy stage and species specificities under the influence of estrogen, progesterone and other growth factor(s). Conceptus also synthesizes components of the IGF system beginning from a period between 2-cell and blastocyst stages. Maternal IGFs are utilized by both maternal and conceptal tissues; conceptus-derived growth factors are believed to be taken up primarily by conceptus. IGFs enhance the development of both maternal and conceptal compartments in a wide range of biological processes. They stimulate proliferation and differentiation of endometrial cells and placental precursor cells including decidual transformation from stromal cells, placental formation and the synthesis of some steroid and protein hormones by differentiated endometrial cells or placenta. It is also well-documented in a number of experimental settings that both IGFs stimulate preimplantation embryo development. In slight contrast to these, prenatal mice carrying a null mutation of IGF and /or IGF receptor gene do not exhibit any apparent growth retardation until after implantation. Reason (s) for this discrepancy between the knock-out result and the in vitro ones, however, is not known. IGFBPs, in general, are believed to inhibit IGF action within the materno-conceptal unit, thereby allowing endometrial stromal cell differentiation as well as dampening ex cessive placental invasion into maternal tissue. There is evidence, however, indicating that IGFBP can enhance IGF action depending on environrnental conditions perhaps by directioning IGF ligand to the target cell. There is also a third possibility that certain IGFBPs and their proteolytic fragments may have their own biological activities independent of the IGF. In addition to IGFBPs, IGFBP proteases including those found within the uterine tissue or lumen are thought to enhance IGF bioavailability by degrading their substrates without affecting their bound ligand. In this regard, preliminary results in early pregnant pigs suggest that a partially characterized IGFBP protease activity in uterine luminal fluid enhances intrauterine IGF bioavailability during conceptus morphological development. In summary, a number of in vitro results indicate that IGFs stimulates the development of the rnaterno-conceptal unit during the periimplantation period. IGFBPs appear to inhibit IGF action by sequestering their ligands, whereas IGFBP proteases are thought to enhance intrauterine bioavailability of IGFs. Much is remaining to be clarified, however, regarding the roles of the individual IGF system components. These include in vivo evidence for the role of IGFs in early conceptus development, identification of IGF-regulated genes and their functions, specific roles for individual IGFBPs, identification and characterization of IGFBP proteases. The intrauterine IGF club house thus will be paying a lot of attention to forthcoming results in above and other areas, with its door wide-open!

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Phospholipase C-β3 Mediates the Thrombin-induced Ca2+ Response in Glial Cells

  • Hwang, Jong-Ik;Shin, Kum-Joo;Oh, Yong-Seok;Choi, Jung-Woong;Lee, Zee-Won;Kim, Daesoo;Ha, Kwon-Soo;Shin, Hee-Sup;Ryu, Sung Ho;Suh, Pann-Ghill
    • Molecules and Cells
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    • v.19 no.3
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    • pp.375-381
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    • 2005
  • Phospholipase C-${\beta}$ (PLC-${\beta}$) hydrolyses phosphatidylinositol 4,5-bisphosphate and generates inositol 1,4,5-trisphosphate in response to activation of various G protein-coupled receptors (GPCRs). Using glial cells from knock-out mice lacking either PLC-${\beta}1$ [PLC-${\beta}1$ (-/-)] or PLC-${\beta}3$ [PLC-${\beta}3$ (-/-)], we examined which isotype of PLC-${\beta}$ participated in the cellular signaling events triggered by thrombin. Generation of inositol phosphates (IPs) was enhanced by thrombin in PLC-${\beta}1$ (-/-) cells, but was negligible in PLC-${\beta}3$ (-/-) cells. Expression of PLC-${\beta}3$ in PLC-${\beta}3$ (-/-) cells resulted in an increase in pertussis toxin (PTx)-sensitive IPs in response to thrombin as well as to PAR1-specific peptide, while expression of PLC-${\beta}1$ in PLC-${\beta}1$ (-/-) cells did not have any effect on IP generation. The thrombin-induced $[Ca^{2+}]_i$ increase was delayed and attenuated in PLC-${\beta}3$ (-/-) cells, but normal in PLC-${\beta}1$ (-/-) cells. Pertussis toxin evoked a delayed $[Ca^{2+}]_i$ increase in PLC-${\beta}3$ (-/-) cells as well as in PLC-${\beta}1$ (-/-) cells. These results suggest that activation of PLC-${\beta}3$ by pertussis toxin-sensitive G proteins is responsible for the transient $[Ca^{2+}]_i$ increase in response to thrombin, whereas the delayed $[Ca^{2+}]_i$ increase may be due to activation of some other PLC, such as PLC-${\beta}4$, acting via PTx-insensitive G proteins.

Genotoxicity and Anti-Oxidative Effectiveness Study of Functional Food Additive Containing Astaxanthin (Astaxanthin 함유 기능성 식품소재의 유전독성 및 항산화능 검사)

  • Kim, Jun-Sung;Park, Jin-Hong;Jin, Hua;Cho, Hyun-Sun;Hwang, Soon-Kyung;Nah, Woon-Seong;Kang, Hwan-Goo;An, Gil-Hwan;Cho, Myung-Haing
    • Toxicological Research
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    • v.22 no.4
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    • pp.381-390
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    • 2006
  • Astaxanthine is a pigment that belongs to the family of the xanthophylls, the oxygenated derivatives of carotenoids whose synthesis in plants derives from lycopene. Astaxanthine is also a carotenoid widely used in salmonid and crustacean aquaculture to provide the pink color characteristic of that. Recent study reported that astaxanthine has the role as a detoxicant against the free radicals. On our study, we estimated the genotoxicity in ICR mice and possibility as antioxidant reagents of mutant Phaffia rhodozyma strain over expressing the astaxanthine by gamma-lay and carophyll pink including astaxanthine in apoE knock out mice, respectively. In our study, we administered Phaffia rhodozyma (2 mg and 3 mg) and carophyll pink for 4 and 8 week. The clinical sign and mortality were not detected compared with control groups. In the mutant frequency of hprt gene and chromosome aberration in splenic cells, there was not detected abnormality. There was not critical change in hematological and serum biochemical test compared to control. In expression level of repair enzyme, increase of catalase were detected and increase of expression level of Nrf-2 was detected in Phaffia rhodozyma (3 mg) and carophyll pink in 8 week treated group. In GSH level, the group of treated with Phaffia rhodozyma (3 mg) showed the increase of the GSH. In conclusion, mutant Phaffia rhodozyma and caphyll pink may be applied to the effective food additives to reduce the free radical.

Effects of different culture systems on the culture of prepuberal buffalo (Bubalus bubalis) spermatogonial stem cell-like cells in vitro

  • Li, Ting-Ting;Geng, Shuang-Shuang;Xu, Hui-Yan;Luo, Ao-Lin;Zhao, Peng-Wei;Yang, Huan;Liang, Xing-Wei;Lu, Yang-Qing;Yang, Xiao-Gan;Lu, Ke-Huan
    • Journal of Veterinary Science
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    • v.21 no.1
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    • pp.13.1-13.14
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    • 2020
  • Currently, the systems for culturing buffalo spermatogonial stem cells (SSCs) in vitro are varied, and their effects are still inconclusive. In this study, we compared the effects of culture systems with undefined (foetal bovine serum) and defined (KnockOut Serum Replacement) materials on the in vitro culture of buffalo SSC-like cells. Significantly more DDX4- and UCHL1-positive cells (cultured for 2 days at passage 2) were observed in the defined materials culture system than in the undefined materials system (p < 0.01), and these cells were maintained for a longer period than those in the culture system with undefined materials (10 days vs. 6 days). Furthermore, NANOS2 (p < 0.05), DDX4 (p < 0.01) and UCHL1 (p < 0.05) were expressed at significantly higher levels in the culture system with defined materials than in that with undefined materials. Induction with retinoic acid was used to verify that the cultured cells maintained SSC characteristics, revealing an SCP3+ subset in the cells cultured in the defined materials system. The expression levels of Stra8 (p < 0.05) and Rec8 (p < 0.01) were significantly increased, and the expression levels of ZBTB16 (p < 0.01) and DDX4 (p < 0.05) were significantly decreased. These findings provided a clearer research platform for exploring the mechanism of buffalo SSCs in vitro.