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Effects of Oviductal Fluid, Culture Media and Zona Pellucida Removal on the Development of Porcine Embryos by Nuclear Transfer

  • Zhang, Y.H.;Song, E.S.;Kim, E.S.;Cong, P.Q.;Lee, S.H.;Lee, J.W.;Yi, Y.J.;Park, Chang-Sik
    • Asian-Australasian Journal of Animal Sciences
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    • v.22 no.7
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    • pp.962-968
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    • 2009
  • The aim of this study was to compare the effects of oviductal fluid, porcine zygote medium (PZM)-3, PZM-4 and PZM-5, and modified PZM-5 culture media, and determine the effects of zona pellucida (ZP) removal on the development of nuclear transfer (NT) embryos. There were no significant differences in the rates of fusion and cleavage among the five different oviductal fluid concentrations. However, the rates of blastocyst formation and the cell numbers per blastocyst were high in the embryos at the 14 and 28 $\mu{g}$/ml concentrations of oviductal fluid compared to the 0, 56 and 100 $\mu{g}$/ml concentrations. The rates of cleavage and blastocyst formation, and the cell numbers per blastocyst were higher in the PZM-3, PZM-5 and modified PZM-5 media than in the PZM-4 medium. However, there were no significant differences in the fusion rates of oocytes among the four culture media. The cell numbers per blastocyst in the embryos without ZP were significantly greater than those with ZP. However, there were no significant differences in the rates of fusion, cleavage and blastocyst formation between the embryos with and without ZP. In conclusion, we improved blastocyst development and the quality of NT embryos by replacing PVA with 3 mg/ml of BSA in PZM-5 medium and supplementing the PZM-5 medium with 14 $\mu{g}$/ml oviductal fluid. The NT embryos produced by the zona-free NT method had a high rate of blastocyst formation in the modified PZM-5 medium.

Inhibition of Growth and Induction of Differentiation of SMMC-7721 Human Hepatocellular Carcinoma Cells by Oncostatin M

  • Kong, N.;Zhang, X.M.;Wang, H.T.;Mu, X.P.;Han, H.Z.;Yan, W.Q.
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.2
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    • pp.747-752
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    • 2013
  • Oncostatin M (OSM) is a multifunctional cellular regulator acting on a wide variety of cells, which has potential roles in the regulation of gene activation, cell survival, proliferation and differentiation. Previous studies have shown that OSM can induce morphological and/or functional differentiation and maturation of many tumor cells. However, the action of OSM on the induction of differentiation of human hepatocellular carcinoma (HCC) has not been reported. Here, we investigated the effects of different concentrations of OSM on human HCC cell line SMMC-7721 growth, proliferation, cell cycling, apoptosis and differentiation in vitro. Cell growth was determined via MTT assay, proliferation by cell cycle analysis, apoptosis by flow cytometry, morphology by transmission electronic microscopy, and cell function by detection of biochemical markers. Our results demonstrated that OSM strongly inhibited the growth of SMMC-7721 cells in a dose-dependent manner, associated with decreased clonogenicity. Cell cycle analysis revealed a decreased proportion of cells in S phase, with arrest at G0/G1. The apotosis rate was increased after OSM treatment compared to the control. These changes were associated with striking changes in cellular morphology, toward a more mature hepatic phenotype, accompanied by significant reduction of the expression of AFP and specific activity of ${\gamma}$-GT, with remarkable increase in secretion of albumin and ALP activity. Taken together, our findings indicate that OSM could induce the differentiation and reduce cell viability of SMMC-7721 cells, suggesting that differentiation therapy with OSM offers the opportunity for therapeutic intervention in HCC.

MicroRNA-122 Promotes Proliferation, Invasion and Migration of Renal Cell Carcinoma Cells Through the PI3K/Akt Signaling Pathway

  • Lian, Ji-Hu;Wang, Wei-Hua;Wang, Jia-Qiang;Zhang, Yu-Hong;Li, Yi
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.9
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    • pp.5017-5021
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    • 2013
  • Objective: MicroRNAs (miRNAs) are a small class of non-coding, single-stranded RNAs with a critical role in genesis and maintenance of renal cancer mainly through binding to 3'-untranslated regions (3'UTR) of target mRNAs, which causes a block of translation and/or mRNA degradation. The aim of the present study was to investigate the potential effects of miR-122 in human renal cell carcinomas. Methods: The expression level of miR-122 was quantified by qRT-PCR. MTT, colony formation, invasion and migration assays were used to explore the potential functions of miR-122 in human renal cell carcinoma cells. Results: Cellular growth, invasion and migration in two A498 and 786-O cells were significantly increased after miR-122 transfection. Further experiments demonstrated that overexpression of miR-122 resulted in the increase of phospho-Akt (Ser473) and phospho-mTOR (Ser2448), then activation of mTOR targets, p70S6K and 4E-BP1. Conclusions: The up-regulation of miR-122 may play an important role in the progress of renal cancer through activating PI3K/Akt signal pathway and could be a potential molecular target for anti-cancer therapeutics.

Salinity Tolerance of Blackgram and Mungbean: I. Dry Matter Accumulation in Different Plant Parts

  • Karim, M.A.;Raptan, P.K.;Hamid, A.;Khaliq, Q.A.;Solaiman, A.R.M.;Ahmed, J.U.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.5
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    • pp.380-386
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    • 2001
  • Dry matter(DM) accumulation in different plant parts of two Vigna spp., blackgram(Vigna mungo) and mungbean(Vigna radiata), was compared at different levels of salinity. Two vaarieties of each of blackgram (Barimash-1 and Barimash-2) and mungbean(Barimung-3 and Barimung-4) were grown with 50, 75 and 100mM NaCl solutions and tap water as a control till maturity. The DM accumulation in all plant parts of the two crops devreased with the increasing salinity levels. The reducation was severe in mungbean compared to blackgram. On an average mungbean produced only 3% grain yield compared to 37% in blackgram at 100mM NaCl. The salinity induced growth reduction was relatively less in Barimash-2 than that in Barimash-1. In mungbean, the relative DM production of Barimung-3 was greater than Barimung-4. The extent of biomass reducation due to salinity in different plant parts was not similar. At maturity the rank of biomass accumulation (at 100 mM NaCl) in different plant parts of blackgram was in decreasing order by seeds pod$^{-1}$ (97%), branch plant$^{-1}$ (88%), 1000-grain weight (79%), plant height(72%), pods plant$^{-1}$ (50%), leaf weight and root mass(both 49%) and stem weight (48%). In mungbean, the rank was in decreasing order by 1000-grain weight (57%), leaf weight (54%), plant height (52%), seeds pod$^{-1}$ (50%), branch plant$^{-1}$ (41%), root weight (34%), stem weight (24%) and pods plant$^{-1}$ (6%). Therefore, salinity reduced grain yield more than straw and roots of the Vignaq spp., and blackgram is relatively more salt-tolerant than mungbean.

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Transcriptional Activity of an Estrogen Receptor β Subtype in the Medaka Oryzias dancena

  • Maeng, Sejung;Yoon, Sung Woo;Kim, Eun Jeong;Nam, Yoon Kwon;Sohn, Young Chang
    • Development and Reproduction
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    • v.23 no.4
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    • pp.333-344
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    • 2019
  • In vertebrate reproductive system, estrogen receptor (ER) plays a pivotal role in mediation of estrogenic signaling pathways. In the present study, we report the cDNA cloning, expression analysis, and transcriptional activity of ERβ1 subtype from medaka Oryzias dancena. The deduced O. dancena ERβ1 (odERβ1; 519 amino acids) contained six characteristic A/B to E/F domains with very short activation function 2 region (called AF2). A phylogenetic analysis indicated that odERβ1 was highly conserved among teleost ERβ1 subgroup. A conventional RT-PCR revealed that the odERβ1 transcripts were widely distributed in the multiple tissues, the ovary, brain, gill, intestine, kidney, and muscle. Further, the relatively higher odERβ1 expressions in the ovary and brain were clearly reproduced in RT-qPCR assay. When HA-fused odERβ1 expression vector was transfected into HEK293 cells, an immunoreactivity for odERβ1 was mainly detected in the nucleus part. Finally, an estrogen responsive element driven luciferase reporter assays demonstrated that the transcriptional activity of odERβ1 significantly increased by estradiol-17β (E2) in a dose dependent manner (p<0.05). However, fold-activation of odERβ1 in the presence of E2 was markedly weak, when it compared with those of O. latipes ERβ1. Taken together, these data suggest that odERβ1 represents a functional variant of teleost ERβ subtype and provides a basic tool allowing future studies examining the function of F domain of ERβ1 subtype and expanding our knowledge of ERβ evolution.

In-Flight Simulation for the Evaluation of Flight Control Law (비행제어계 평가를 위한 항공기 공중모의 비행시험)

  • Go,Jun-Su;Lee,Ho-Geun;Lee,Jin-Yeong
    • Journal of the Korean Society for Aeronautical & Space Sciences
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    • v.31 no.10
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    • pp.79-88
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    • 2003
  • The paper presented here covers the work associated with the flight control law design, ground based and in flight simulation and handling qualities assessment of the Fly-by-Wire type Aircraft (FBWA). The FBWA configurations are of the same generic form of the Korean advanced trainer. The normal acceleration (Nz) and pitch rate (q) feedback control system is employed for longitudinal axis and roll rate (p) and lateral acceleration (Ny) feedback flight control law is developed in lateral/ directional axis. The flight tests for the FBW A dynamics evaluation were executed for the target aircraft (FBWA) on the IFS (In-Flight-Simulator) aircraft . The test results showed that Level 1 handling qualities for the most unstable flight regime and Level 1/2 for the landing approach flight regime were achieved. And the designed FBWA flight control law has revealed acceptable CHR (Cooper-Harper handling qualities Ratings).

Analysis of Squalene Synthase Expression During the Development of Ganoderma lucidum

  • Zhao, M.W.;Zhong, J.Y.;Liang, W.Q.;Wang, N.;Chen, M.J.;Zhang, D.B.;Pan, Y.J.;Jong.S.C.
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.116-120
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    • 2004
  • The medicinal properties of Ganoderma lucidum have been recognized in China for many centuries. Active pharmaceutical components include triterpenes. To elucidate the molecular regulation of triterpene biosynthesis in this mushroom, a 57-base pair DNA fragment encoding the fourth conserved domain SQ-4 (SMGLFLQKTNIIRDYNEDL) of squalene synthase was synthesized and cloned into the expression vector pET-32a(+). The recombinant fusion protein induced by IPTG (isopropyl-$\beta$-D-thiogalactopyranoside) was overexpressed in the Escherichia coli. Using the purified recombinant fusion protein of 20.9 kDa, a specific polyclonal antibody was obtained from immunized rabbit. Expression of squalene synthase at different development stages of Ganoderma lucidum was analyzed.

A New Species of Hyphomycetes, Aspergillus coreanus sp.nov.,Isolated from Traditional Korean Nuruk

  • Yu, Tae-Shick;Yeo, Soo-Hwan;Kim, Hyun-Soo
    • Journal of Microbiology and Biotechnology
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    • v.14 no.1
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    • pp.182-187
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    • 2004
  • Strain NR $15-1^T$ isolated from traditional Korean Nuruk is described as a new species and named as Aspergillus coreanus NR $15-1^T$ sp. novo Strain NR $15-1^T$ grew rapidly to form yellow-green colonies whose surfaces were velvety on Czapek solution agar. Conidial heads were yellow to light and elliptical, whereas the conidiophore was colorless and typically long. In addition, vesicles were from flask-shaped to globose, and sterigmata are uniseriate. Conidia were spherical and deep yellow-green, and their surfaces were lightly roughened. The G+C content of strain NR $15-1^T$ was 51 mol% and strain NR $15-1^T$contained a dihydrogenated ubiquinone with Q9 (94.9%) as a major quinone. The nucleotide sequences of strain NR $15-1^T$ in the two Internal Transcribed Spacers (ITS 1 and 2) and 5.8S rDNA showed highest similarity when compared with that of A. tubingensis and A. phoenicis NRRL $365^T$. However, based on morphological and chemotaxonomic characteristics, this strain was different from A. tubingensis and A. phoenicis NRRL $365^T$. On the basis of the data presented, it is proposed that strain NR $15-1^T$ should be placed in the genus Aspergillus as a new species, Aspergillus coreanus sp. novo Therefore, the type strain of the new species is strain NR $15-1^T$ (=KCTC 18075P^T,=KCCM 80006^T$.

Molecular Cloning and Nucleotide Sequence of the Gene Encoding Fusion(F) Protein of the Thermostable Newcastle Disease Virus Isolated from a Diseased Pheasant (꿩에서 분리된 Newcastle Disease Virus 내열성주 (CBP)의 Fusion(F) 유전자 클론닝과 염기서열 분석)

  • Chang, Kyung-Soo;Jun, Moo-Hyung;Song, Hee-Jong;Kim, Kui-Hyun;Park, Jong-Hyeon
    • The Journal of Korean Society of Virology
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    • v.28 no.3
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    • pp.233-245
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    • 1998
  • The gene encoding F protein of CBP-1 strain, a heat-stable Newcastle disease virus (NDV) isolated from the diseased pheasants in Korea, was characterized by reverse transcription-polymerase chain reaction (RT-PCR), nucleotide and amino acid sequences. Virus RNA was prepared from the chorioallatoic fluid infected with NDV CBP-1 virus and cDNA was amplified by RT-PCR, cloned and sequenced to analyze. The PCR was sensitive as to detect the virus titer above $2^5$ hemagglutination unit. 1.7kb (1,707bp) size of the cDNA was amplified and cloned into BamHI site of pVL1393 Baculo transfer vector. The nucleotide sequences for F protein were determined by dye terminator cyclic sequencing using four pairs of primers, and 553 amino acid sequences were predicted. In comparison of the nucleotide sequence of F gene of CBP-1 with those of other NDV strains, the homology revealed 88.8%, 98.5% and 98.7% with Kyojungwon (KJW), Texas GB and Beaudette C strains, respectively. As the deduced 553 amino acid sequences of F protein of CBP-1 were compared with those of other NDV strains, the homology appeared 89.9%, 98.7% and 98.9% with KJW, Texas GB and Beaudette C strains, respectively. The putative protease cleavage site (112-116) was R-R-Q-K-R, indicating that CBP-1 strain is velogenic type. The amino acid sequences include 6 sites of N-asparagine-linked glycosylation and 13 cysteine residues. These data indicate that the genotype of CBP-1 strain is more closely associated with the strains of Texas GB and Beaudette C than KJW strain.

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Poncirin Inhibits Osteoclast Differentiation and Bone Loss through Down-Regulation of NFATc1 In Vitro and In Vivo

  • Chun, Kwang-Hoon;Jin, Hyun Chul;Kang, Ki Sung;Chang, Tong-Shin;Hwang, Gwi Seo
    • Biomolecules & Therapeutics
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    • v.28 no.4
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    • pp.337-343
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    • 2020
  • Activation of osteoclast and inactivation of osteoblast result in loss of bone mass with bone resorption, leading to the pathological progression of osteoporosis. The receptor activator of NF-κB ligand (RANKL) is a member of the TNF superfamily, and is a key mediator of osteoclast differentiation. A flavanone glycoside isolated from the fruit of Poncirus trifoliata, poncirin has anti-allergic, hypocholesterolemic, anti-inflammatory and anti-platelet activities. The present study investigates the effect of poncirin on osteoclast differentiation of RANKL-stimulated RAW264.7 cells. We observed reduced formation of RANKL-stimulated TRAP-positive multinucleated cells (a morphological feature of osteoclasts) after poncirin exposure. Real-time qPCR analysis showed suppression of the RANKL-mediated induction of key osteoclastogenic molecules such as NFATc1, TRAP, c-Fos, MMP9 and cathepsin K after poncirin treatment. Poncirin also inhibited the RANKL-mediated activation of NF-κB and, notably, JNK, without changes in ERK and p38 expression in RAW264.7 cells. Furthermore, we assessed the in vivo efficacy of poncirin in the lipopolysaccharide (LPS)-induced bone erosion model. Evaluating the micro-CT of femurs revealed that bone erosion in poncirin treated mice was markedly attenuated. Our results indicate that poncirin exerts anti-osteoclastic effects in vitro and in vivo by suppressing osteoclast differentiation. We believe that poncirin is a promising candidate for inflammatory bone loss therapeutics.