• Title/Summary/Keyword: SH2D4A

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SH2D4A regulates cell proliferation via the ERα/PLC-γ/PKC pathway

  • Li, Tingting;Li, Wei;Lu, Jingyu;Liu, Hong;Li, Yinghui;Zhao, Yanyan
    • BMB Reports
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    • v.42 no.8
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    • pp.516-522
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    • 2009
  • SH2D4A, comprising a single SH2 domain, is a novel protein of the SH2 signaling protein family. We have previously demonstrated SH2D4A is expressed ubiquitously in various tissues and is located in the cytoplasm. In this study we investigated the function of SH2D4A in human embryonic kidney (HEK) 293 cells using interaction analysis, cell proliferation assays, and kinase activity detection. SH2D4A was found to directly bind to estrogen receptor $\alpha$ (ER$\alpha$), and prevent the recruitment of phospholipase C-$\gamma$ (PLC-$\gamma$) to ER$\alpha$. Moreover, we observed its inhibitory effects on estrogen-induced cell proliferation, involving the protein kinase C (PKC) signaling pathway. Together, these findings suggested that SH2D4A inhibited cell proliferation by suppression of the ER$\alpha$/PLC-$\gamma$/PKC signaling pathway. SH2D4A may be useful for the development of a new anti-cancer drug acting as an ER signaling modulator.

Effect of Embryo Maturity and Medium on Callus Formation and Plant Regeneration from Immature Embryo of Eleutherococcus senticosus (가시오갈피 미숙배로부터 Callus 형성 및 식물체 재분화에 미치는 배의 성숙정도 및 배지의 효과)

  • 유창연
    • Korean Journal of Plant Resources
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    • v.10 no.2
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    • pp.122-127
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    • 1997
  • This study was conducted to establish mass propagation system from the tissue culture using immature embroys in Eleutherococus senticosus. Immature embroys from seeds were removed under the microscope and placed on modified SH and WPM medium containing several plant growth regulators. The calli were well formed on media containing 1mg/l of 2,4-D on modified SH medium and 1mg/l of 2,4-D and 3mg/l of TDZ on WPM medium. Shoot regeneration was better on modified SH or WPM medium with combination of high concentration of TDZ and low concentration of 2,4-D. Treatment of 2,4-D alone was better than treatment of TDZ alone in callus induction on modified SH medium but plant regeneration reversed. Treatment of 2.4-D and TDZ combination was better than treatment of 2,4-D alone in callus induction on WPM medium. The results of callus formation and shoot regeneration on WPM media differed to those of SH media. The rate of callus formation was nearly 83% when 2,4-D was added to SH medium on concentration of 1mg/l. The rate of callus formation was nearly 38% when combination treatment of 2,4-D 1mg/l and TDZ 3mg/l was added to WPM medium. Also, plant regeneration differed depending on the mature degree of immature embryo.

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Circ_UBE2D2 Attenuates the Progression of Septic Acute Kidney Injury in Rats by Targeting miR-370-3p/NR4A3 Axis

  • Huang, Yanghui;Zheng, Guangyu
    • Journal of Microbiology and Biotechnology
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    • v.32 no.6
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    • pp.740-748
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    • 2022
  • As circ_UBE2D2 has been confirmed to have targeted binding sites with multiple miRNAs involved in septic acute kidney injury (SAKI), efforts in this study are directed to unveiling the specific role and relevant mechanism of circ_UBE2D2 in SAKI. HK-2 cells were treated with lipopolysaccharide (LPS) to construct SAKI model in vitro. After sh-circ_UBE2D2 was transfected into cells, the transfection efficiency was detected by qRT-PCR, cell viability and apoptosis were determined by MTT assay and flow cytometry, and expressions of Bcl-2, Bax and Cleaved-caspase 3 were quantified by western blot. Target genes associated with circ_UBE2D2 were predicted using bioinformatics analysis. After the establishment of SAKI rat model, HE staining and TUNEL staining were exploited to observe the effect of circ_UBE2D2 on tissue damage and cell apoptosis. The expression of circ_UBE2D2 was overtly elevated in LPS-induced HK-2 cells. Sh-circ_UBE2D2 can offset the inhibition of cell viability and the promotion of cell apoptosis induced by LPS. Circ_UBE2D2 and miR-370-3p as well as miR-370-3p and NR4A3 have targeted binding sites. MiR-370-3p inhibitor reversed the promoting effect of circ_UB2D2 silencing on viability of LPS-treated cells, but shNR4A3 neutralized the above inhibitory effect of miR-370-3p inhibitor. MiR-370-3p inhibitor weakened the down-regulation of NR4A3, Bax and Cleaved caspase-3 and the up-regulation of Bcl-2 induced by circ_UB2D2 silencing, but these trends were reversed by shNR4A3. In addition, sh-circ_UBE2D2 could alleviate the damage of rat kidney tissue. Circ_UBE2D2 mitigates the progression of SAKI in rats by targeting miR-370-3p/NR4A3 axis.

Characterization of Cysteine Residues in Cabbage Phospholipase D by Sulfhydryl Group Modifying Chemicals (설프히드릴 변형 화합물질들에 의한 양배추 포스포리파제 D의 시스테인 잔기의 특성)

  • Go, Eun-Hui
    • Journal of the Korean Chemical Society
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    • v.50 no.5
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    • pp.362-368
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    • 2006
  • SH group modifying chemicals were used to characterize the eight cysteine residues of cabbage PLD. 5,5-dithiobis(2-nitrobenzoate)(DTNB) was used to titrate the SH group of cysteine residues . Based on the optical density at 412nm due to the reduced DTNB, 4 SH groups are found to be present in a native PLD while 8 SH groups in the denatured PLD whose tertiary structure was perturbed by 8M urea. The results imply that among the 8 cysteine residues of PLD, the half(4) are exposed on the surface whereas the other half are present at the interior of the enzyme tertiary structure. The PLD was inactivated by SH modifying reagents such as p-chloromercuribenzoate(PCMB), iodoacetate, iodoacetamide, and N-ethylmaleimide. At the addition of dithiothreitol(DTT) only the PCMB inhibited PLD activity was recovered reversibly. The micro-environment of the exposed SH group of cysteine residues was examined with various disulfide compounds with different functional groups and we found that anionic or neutral disulfides appear to be more effective than the positively charged cystamine for inactivating the PLD activity. The effect of redox state of cysteine residues on the PLD activity was further explored with H2O2. The oxidation of SH groups by H2O2 inhibited the PLD activity more than 70%, which was mostly recovered by DTT. From these results, we could confirm chemically that all the cysteine residues of PLD are present as in their reduced SH forms and the 4 SH groups exposed on the surface of the enzyme may play important roles in the regulation of PLD activity.

Mass Propagation of Dicentra spectabilis L. Lemaire Through In vitro Suspension Culture (현탁배양을 통한 금낭화(Dicentra spectabilis L. Lemaire)의 대량증식)

  • Lee, Kang-Seop;Sim, Ock-Kyeong;Shin, Jeong-Sun;Choi, Yong-Eui;Kim, Ee-Yup
    • Journal of Plant Biotechnology
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    • v.31 no.2
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    • pp.121-126
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    • 2004
  • Bleeding heart (Dicentra spectabilis L. Lemaire) is one of the most valuable wild flower in Korea. This work was conducted for the mass production of somatic embryos through suspension culture and more effective plant regeneration system in Dicentra spectabilis. High-frequency embryogenic callus proliferation was achieved in SH liquid medium supplemented with 1 mg/L 2,4-D. Half-strength SH medium was suitable concentration for somatic embryo induction and germination. About 5,000 embryos were produced per 250$m\ell$ flask after 4 weeks of culture. Germination rate of somatic embryos was decreased when GA$_3$ was added in medium. The plantlets showed a 58% survival rate when transferred to pots after 1 month of culture. The results indicate that micropropagation procedure via somatic embryogenesis can be applied for an efficient mass propagation of Dicentra spectabilis.

Effect of Replacing Corn and Wheat Bran With Soyhulls in Lactation Cow Diets on In Situ Digestion Characteristics of Dietary Dry Matter and Fiber and Lactation Performance

  • Meng, Qingxiang;Lu, Lin;Min, Xiaomei;McKinnon, P.J.;Xiong, Yiqiang
    • Asian-Australasian Journal of Animal Sciences
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    • v.13 no.12
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    • pp.1691-1698
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    • 2000
  • An in situ digestion trial (Experiment 1) and a lactation trial (Experiment 2) were conducted to determine the effects of replacing corn and wheat bran with soyhulls (SH) in lactating dairy cow diets on the extent and kinetics of digestion of DM and NDF, and lactation performance. In experiment 1, five mixed feeds consisting of mixed concentrate and roughages (50:50 on a DM basis) were formulated on isonitrogenous and isoenergetic bases to produce five levels (0, 25, 50, 75 and 100%) of SH replacement for corn and wheat bran. SH had high in situ digestion (92 and 89% for potentially digestible DM and NDF) and fairly fast digestion rate (7.2 and 6.3 %/h for DM and NDF). Increasing level of SH replacement resulted in increased NDF digestibility (linear, p=0.001-0.04) and similar DM digestibility (beyond 12 h incubation, p=0.10-0.41). As level of SH replacement increased, percentage of slowly digestible fraction (b) of DM increased (linear, p=0.03), percentage of rapidly digestible fraction (a) of DM tended to decrease (linear, p=0.14), and DM digestion lag time tended to be longer (linear, p=0.13). Percentage of potentially digestible fraction (a+b) and digestion rate (c) of slowly digestible fraction of dietary DM remained unaltered (p=0.36-0.90) with increasing SH in the diet. Increasing level of SH for replacing corn and wheat bran in the diet resulted in increases in percentages of b (quadratic, p<0.001), a (linear, p=0.08), a+b (quadratic, p=0.001) and a tendency to increase in c for NDF (linear, p<0.19). It was also observed that there was a satisfactory fit of a non-linear regression model to NDF digestion data ($R^2=0.986-0.998$), but a relatively poor fit of the model to DM digestion data ($R^2=0.915-0.968$). In experiment 2, 42 lactating Holstein cows were used in a randomized complete block design. SH replaced corn and wheat bran in mixed concentrates at 0, 25, and 50%, respectively. These mixed concentrates were mixed with roughages and fed ad libitum as complete diets. Replacing corn and wheat bran with SH at 0, 25 and 50% levels did not influence (p=0.56-0.95) DM intakes (18.4, 18.6, and 18.5 kg/d), milk yields (27.7, 28.4 and 27.6 kg/d), 4% fat-corrected-milk (FCM) yields (26.2, 27.6, and 27.3 kg/d) and percentages of milk protein (3.12, 3.17 and 3.18%), milk lactose (4.69, 4.76 and 4.68%) and SNF (8.50, 8.64, and 8.54%). On the other hand, milk fat percentges linearly increased (3.63, 3.85 and 3.90% for SH replacement rates of 0, 25 and 50% in the diet, p=0.08), while feed costs per kg FCM production were reduced.

Somatic Embryogenesis and Plant Regeneration in Leaf Explant Cultures of Gentiana scabra var buergeri (용담(Gentina scabra var. buergeri)의 잎 절편 배양에서 체세포배발생에 의한 식물체 재분화)

  • 방재욱;이미경;정성현
    • Korean Journal of Plant Tissue Culture
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    • v.21 no.4
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    • pp.233-237
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    • 1994
  • Plant regeneration system via somatic embryogenesis in leaf explant cultures of Gentiana scabra var. buergeri has been established. Leaf segments formed calli when cultured on MS medium supplemented with 0.5 mg/L 2,4-D and 2 mg/L BAP After transferred to SH medium supplemented with 0.5 mg/L 2,4-D, 2 mg/L CPA and 0.5 mg/L kinetin, the callus became embryogenic. The embryogenic callus was subcultured every 3 to 4 weeks. Upon transfer onto SH basal medium the embryogenic callus gave rise to numerous somatic embryos, which subsequently developed into plantlets. The regenerated plants were potted in an artificial soil with mixture (peatmoss : pearlite : vermiculite : 2 : 1 : 1) and transplanted to the soil after kept under a high humidity for two weeks. A total of 78 plants out of 105 regenerated plants survived in the soil. Phenotypic variations in height, number of stems and the flowering time were observed in tile regenerated plants. Cytogenetical analyses showed no chromosomal variation.

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Waveform inversion of shallow seismic refraction data using hybrid heuristic search method (하이브리드 발견적 탐색기법을 이용한 천부 굴절법 자료의 파형역산)

  • Takekoshi, Mika;Yamanaka, Hiroaki
    • Geophysics and Geophysical Exploration
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    • v.12 no.1
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    • pp.99-104
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    • 2009
  • We propose a waveform inversion method for SH-wave data obtained in a shallow seismic refraction survey, to determine a 2D inhomogeneous S-wave profile of shallow soils. In this method, a 2.5D equation is used to simulate SH-wave propagation in 2D media. The equation is solved with the staggered grid finite-difference approximation to the 4th-order in space and 2nd-order in time, to compute a synthetic wave. The misfit, defined using differences between calculated and observed waveforms, is minimised with a hybrid heuristic search method. We parameterise a 2D subsurface structural model with blocks with different depth boundaries, and S-wave velocities in each block. Numerical experiments were conducted using synthetic SH-wave data with white noise for a model having a blind layer and irregular interfaces. We could reconstruct a structure including a blind layer with reasonable computation time from surface seismic refraction data.

Effects of Media and Elicitor on Betalain and Phytolaccosides Production in Hairy Root Cultures of Phytolacca esculenta van Houtte (자리공(Phytolacca esculenta van Houtte) 모상근의 Betalain 및 Phytolaccosides 생성에 미치는 배지 및 Elicitor 영향)

  • 양덕조;김용해;윤길영
    • Korean Journal of Plant Tissue Culture
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    • v.25 no.6
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    • pp.507-514
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    • 1998
  • The effects of media and elicitors on betalain, phytolaccoside G and D2 production were examined in the hairy roots of Phytolacca esculenta van Houtte induced by Agrobacterium tumefaciens $A_4$T. Phytolaccoside G and D2 from Phytolacca hairy roots PEH2 were identified by TLC, HPLC, IR, Mass, $^1$H-NMR, and ^(13)C-NMR. Among the culture media tested, SH medium was the best for hairy root growth of hairy roots. White medium was the most suitable medium for betalain production, while MS medium was for phytolaccoside G and D2 production. Although the growth of hairy roots was supped by light (1,000 Lux), the production of betalain, phytolaccoside G and D2 was enhanced by the same light treatment. Addition of elicitors such as NaF, chitosan, and yeast extract to the culture medium increased the content of betalain, phytolaccoside G and D2, suggesting the importance of culture condition for the production of those componds.

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Synthesis and pH-Dependent Micellization of Sulfonamide-Modified Diblock Copolymer

  • Pal Ravindra R.;Kim Min Sang;Lee Doo Sung
    • Macromolecular Research
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    • v.13 no.6
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    • pp.467-476
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    • 2005
  • The main objective of this study was to develop and characterize pH-sensitive biodegradable polymeric materials. For pH-sensitivity, we employed three kinds of moieties: 2-amino-3-(lH-imidazol-4-yl)-propionic acid (H), N-[4-( 4,6-dimethyl-pyrimidin-2ylsulfamoyl)-phenyl]succinamic acid (SM), and 2- {3-[ 4-( 4,6-dimethyl-pyrim­idin- 2-ylsulfamoyl)-phenylcarbamoyl]-propionylamino} -3-(3 H - imidazol-4-yl)-propionic acid (SH). The pH -sensitive diblock copolymers were synthesized by ring opening polymerization and coupling reaction from poly(ethylene glycol) (MPEG), $\varepsilon$-caprolactone (CL), D,L-lactide (LA) and pH-sensitive moieties. The pH-sensitive SH molecule was synthesized in a two-step reaction. The first step involved the synthesis of SHM, a methyl ester derivative of SH, by coupling reaction of SM and L-histidine methyl ester dihydrochloride, whereas the second step involved the hydrolysis of the same. The synthesized SM, SHM and SH molecules were characterized by FTIR, $^{1}H$-NMR and $^{13}C$-NMR spectroscopy, whereas diblock copolymers and pH-sensitive diblock copolymer were characterized by $^{1}H$-NMR and GPC analysis. The critical micelle concentrations were determined at various pH conditions by fluorescence technique using pyrene as a probe. The micellization and demicellization studies of pH-sensitive diblock copolymers were also done at different pH conditions. The pH-sensitivity was further established by acid-based titration and DLS analysis.