• Title/Summary/Keyword: BF-2 cells.

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Antioxidant and growth inhibitory activities of Mesembryanthemum crystallinum L. in HCT116 human colon cancer cells (아이스플랜트의 항산화 및 HCT116 인체 유래 대장암세포 성장억제 활성)

  • Seo, Jin A;Ju, Jihyeung
    • Journal of Nutrition and Health
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    • v.52 no.2
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    • pp.157-167
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    • 2019
  • Purpose: This study examined the antioxidant and cancer cell growth inhibitory activities of an ethanol extract and different solvent fractions of Mesembryanthemum crystallinum L. (ice plant). Methods: The ice plant was freeze-dried, extracted with 99.9% ethanol, and then fractionated with hexane, ethyl acetate, butanol, and water. The total polyphenol content (TPC), total carotenoid content (TCC), 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity (RSA), and ferric reducing antioxidant power (FRAP) were measured. Assays using 2',7'-dichlorofluorescin-diacetate and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide were performed to measure the intracellular reactive oxygen species (ROS) and cell growth, respectively. Annexin V/propidium iodide staining and cell cycle analysis were performed for the detection of apoptosis and cell cycle arrest. Results: TPC, TCC, RSA, and FRAP of the ethanol extract (EE) were 3.7 mg gallic acid equivalent/g, $13.2{\mu}g/g$, 21.0% (at a concentration of 5 mg/mL), and 21.0% (at a concentration of 5 mg/mL), respectively. Among the different solvent fractions, the butanol fraction (BF) showed the highest TPC (5.4 mg gallic acid equivalent/g), TCC ($86.6{\mu}g/g$), RSA (34.9% at 5 mg/mL), and FRAP (80.8% at 5 mg/mL). Treatment of HCT116 human colon cancer cells with EE and BF at concentrations of 250 and $500{\mu}g/mL$ reduced the levels of intracellular ROS. Concomitantly, EE and BF resulted in the dose-dependent inhibition of cell growth (at the concentrations of 125, 250, and $500{\mu}g/mL$ for 24 ~ 48 h) and the induction of apoptosis (at the concentrations of 250 and $500{\mu}g/mL$ for 48 h) in HCT116 cells. An increased G2/M cell population was also found in the BF-treated cells. Conclusion: These results suggest that ice plant possesses antioxidant and growth inhibitory activities in colon cancer cells.

Development of In vitro Fertilized Bovine Embryos in Medium Supplemented with Matrigel (Matrigel이 첨가된 배양액에서 소 체외수정란의 발달)

  • 김동훈;김세웅;이민정;황인선;배성훈;양병철;임기순;성환후;양보석
    • Journal of Animal Science and Technology
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    • v.48 no.6
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    • pp.805-812
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    • 2006
  • The purpose of this study was to investigate the effect of Matrigel on the development of bovine embryos after in vitro fertilization. Bovine embryos were cultured in Ⅰ) SOF+ 0.8% BSA(SOF-B), Ⅱ) SOF+ 0.8% BSA plus 0.8% Matrigel(SOF-M) and III) SOF+0.8% BSA and 10% FBS(SOF-BF). The addition of Matrigel appeared not to increase the proportion of blastocysts (SOF-B, 26.6%; SOF-M, 28.2%; SOF-BF, 26.2%). However, the proportion of hatched blastocysts were significantly increased(P<0.05) by Matrigel(SOF-B, 23.7%; SOF-M, 48.7%; SOF-BF, 18.5%). The means of cell number blastocyst was not significantly different among the treatment groups(SOF-B, 172.7±35.5; SOF-M, 175.1±37.4; SOF-BF, 172.8±38.1). The proportion of apoptotic cells in blastocyst was also found to be not significant among the treatment groups(SOF-B, 3.6±3.2%; SOF-M, 4.3±2.6%; SOF-BF, 4.9±4.3%). In this experiment, Matrigel appeared to support embryonic hatching of bovine embryos. Results suggest that Matrigel, as extracellular matrix components, may be another avenue for formulating more physiological culture system in serum-free culture.

Infectious Bursal Disease-A Review (전염성 훼브리셔스낭병)

  • 이영옥
    • Korean Journal of Poultry Science
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    • v.7 no.2
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    • pp.18-27
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    • 1980
  • Infectious bursal disease, so called Gumboro disease, is found world-wide in areas of intensive poultry farming. The clinical signs of the disease are very indicative, but most infections occur unnoticed due to the age of infection of chicken as well as the degree of virulence of virus affected. Edematous and hemorrhagic lesions in BF at early course of infection and the complete atrophies of BF in later are the most characteristic. The infection is considered highly contagious by direct contact, by fecal material and by contaminated feed and water. The virus is also highly resistant in environment and belongs to Diploma virus with size of 55 to 60nm of Ribovirus group. IBDV grows in embryos, embryonic cells and BF of susceptible chickens. Immune-diffusion using agar gel is the method of a choice to determine IBDV infection in chickens. Maternal immunity is very effective in protecting chickens of critical age when IBDV infection severely damages the function of BF. Immunosuppressive effect of IBDV causes more production losses than direct effects of clinical disease of IBD. Inclusion body hepatitis, infectious anemia and gangrenous dermatitis syndrome are the disease associated with the immunosuppressive condition of chickens.

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Change of Infection Properties of Subcultured Marine Birnavirus in Several Fish Cell Lines (어류 주화세포에서의 계대배양에 의한 해양버나바이러스의 감염특성의 변화)

  • Jung, Sung-Ju
    • Journal of fish pathology
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    • v.11 no.2
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    • pp.89-96
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    • 1998
  • Marine birnavirus (MABV) has wide host range in marine organisms. To clarify various infection properties of MABV in different host species, in vitro study was performed by subculture for 10 passages in several fish cell lines. In CHSE-214, RTG-2 and RSBK-2 cells, the virus produced high yield of virus. Typical CPE with high protein expression was observed in these cells. On the contrary, the virus grown in EPC, FHM and BF-2 cells exhibited no CPE appearance although virus protein was detected. In EPC and FHM cells, the virus titer increased in later passages. The plaque size was distinctly bigger in CHSE-214, RTG-2 and RSBK-2 cells than in other cell lines. The nucleotide sequence of VP2/NS junction region on genome segment A exhibited one specific nucleotide change at 195. The different infection properties in several cell types performed in the present work might reflect in vivo MABV infection in various host species occurring in natural conditions.

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Electrochemical Properties of Trimethylammonium Tetrafluoroborate in Electrochemical Double-Layer Capacitors

  • Lee, Sooyeon;Lee, Kyung Min;Kim, Ketack
    • Journal of Electrochemical Science and Technology
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    • v.13 no.2
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    • pp.254-260
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    • 2022
  • Trimethylammonium tetrafluoroborate (TriMA BF4), consisting of the smallest trialkylammonium ion, was investigated for use in electrochemical double-layer capacitors. Despite the presence of a proton in TriMA+, cycle life tests in acetonitrile (AN) and -butyrolactone (GBL) showed a good capacity retention with a 1.8 V cut-off voltage. The rate of electrolysis of TriMA BF4 in GBL was lower than that in AN because of the lower conductivity in GBL. As a consequence, the cells based on GBL achieved a higher capacitance and longer life than those with AN. TriMA BF4 had a higher conductivity and lower viscosity than the quaternary salt tetraethylammonium tetrafluoroborate in GBL, as well as higher ionic mobility, these factors resulted in a higher rate capability.

Persistent Infection of Marine Birnavirus and its Status of Infection in Cells (잠복감염시의 해양버나바이러스의 세포내에서의 동태)

  • Jung, Sung-Ju;Oh, Myung-Joo
    • Journal of fish pathology
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    • v.15 no.1
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    • pp.9-16
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    • 2002
  • The objective of the study was to clarify the mechanism of persistent infection of marine birnavirus (MABV) in various nonpermissive cell lines. It was observed in CHSE-214, RTG-2 and RSBK-2 that the virus produced at high yield with typical cytopathic effect (CPE). On the contrary, the CPE was not produced in EPC, FHM and BF-2 cells. However amount of virus protein in both permissive and nonpermissive cell lines detected by ELISA was almost the same. Electron microscopy showed virions in permissive cells but not in nonpermissive cells. From the results, it is clear that virus protein and RNA were produced in nonpermissive cells as observed in permissive cells; however, assembly of the virus particles did not occur in nonpermissive cells.

Enhanced 2,5-Furandicarboxylic Acid (FDCA) Production in Raoultella ornithinolytica BF60 by Manipulation of the Key Genes in FDCA Biosynthesis Pathway

  • Yuan, Haibo;Liu, Yanfeng;Lv, Xueqin;Li, Jianghua;Du, Guocheng;Shi, Zhongping;Liu, Long
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.1999-2008
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    • 2018
  • The compound 2,5-furandicarboxylic acid (FDCA), an important bio-based monomer for the production of various polymers, can be obtained from 5-hydroxymethylfurfural (HMF). However, efficient production of FDCA from HMF via biocatalysis has not been well studied. In this study, we report the identification of key genes that are involved in FDCA synthesis and then the engineering of Raoultella ornithinolytica BF60 for biocatalytic oxidation of HMF to FDCA using its resting cells. Specifically, previously unknown candidate genes, adhP3 and alkR, which were responsible for the reduction of HMF to the undesired product 2,5-bis(hydroxymethyl)furan (HMF alcohol), were identified by transcriptomic analysis. Combinatorial deletion of these two genes resulted in 85.7% reduction in HMF alcohol formation and 23.7% improvement in FDCA production (242.0 mM). Subsequently, an aldehyde dehydrogenase, AldH, which was responsible for the oxidation of the intermediate 5-formyl-2-furoic acid (FFA) to FDCA, was identified and characterized. Finally, FDCA production was further improved by overexpressing AldH, resulting in a 96.2% yield of 264.7 mM FDCA. Importantly, the identification of these key genes not only contributes to our understanding of the FDCA synthesis pathway in R. ornithinolytica BF60 but also allows for improved FDCA production efficiency. Moreover, this work is likely to provide a valuable reference for producing other furanic chemicals.

Development of DNA Vaccine Against Red Sea Bream Iridovirus (RSIV)

  • PARK SO-JIN;SEO HYO-JIN;SON JEONG HWA;KIM HYOUNG-JUN;KIM YUN-IM;KIM KI-HONG;NAM YOON-KWON;KIM SUNG-KOO
    • Journal of Microbiology and Biotechnology
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    • v.15 no.4
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    • pp.873-879
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    • 2005
  • Red sea bream iridovirus (RSIV) obtained from infected rock bream was propagated by Bluegill fry-2 (BF-2) cell culture. The virus titer was determined as $10^{5.5}\;TCID_{50}/ml$ on confluent BF-2 cell monolayers. The integrin binding site of ORF 055L of infectious spleen and kidney necrosis virus (ISKNV) was selected for the construction of a primer to obtain the RSIV ORF 055L gene. The genes were amplified using RSIV gene lyzate by PCR. The homologies of the ORF 055L sequence of RSIV with ISKNV and rock bream iridovirus (RBIV) were approximately $96\%$ and $100\%$, respectively. DNA vaccine was constructed by cloning the ORF 055L of RSN into pcDNA 3.1 (+), containing a cytomegalovirus (CMV) promoter. For antibody production, pcDNA-055 DNA vaccine was injected to BALB/c mice. The production of antibodies against pcDNA-055 DNA vaccine was confirmed by the Western blot analysis. The antibodies produced by the pcDNA-055 DNA vaccine showed efficacy to neutralize the RSIV in the neutralization test in BF-2 cell culture.

Analysis of Fish Expression Vectors for Construction of Two MARs Expression Vector System in Fish Cell Line

  • Lim, Hak-Seob;Park, Jin-Young;Hwnag, Jee-Hwang;Kim, Moo-Sang;Lee, Hyung-Ho
    • Journal of Aquaculture
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    • v.13 no.1
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    • pp.29-37
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    • 2000
  • In previously study we isolated several fish matrix attachment regions (MARs) capable of replicating the plasmid by itself. In this study we construct a fish expression vector pBaEGFP(+) containing mud loach ${\beta}$-actin promoter EGFP as reporter gene and SV40 signal. To analyze the effects of the fish expression vector respectively. The fish ARS containing constructs pBaEGFP(+)-ARSs were transfected cells with pBaEGFP(+)-ARS101 and pBaEGFP(+)-ARS223 reduced 10 days to 25 days and then was constant to 30 days after transfection while that of the control vector without ARS element was basal level. The intensity of both constructs showed about 30fold of the intensity compared with the control vector on 30days after transfection individually .E. coli back-transformation analysis shows that pBaEGFP(+)-ARS223 and pBaEGFP(+)-ARS905 maintain in episomal state at least 30 days after transfection. The result indicates that both may be able to replicate the vector in BF-2 cell. Therefore the matrix-attached ARSs enhancing expression of the reporter gene might be useful as a component o the expression vector for transgenic studies.

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Estimates of Genetic Correlations between Production and Semen Traits in Boar

  • Oh, S.H.;See, M.T.;Long, T.E.;Galvin, J.M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.2
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    • pp.160-164
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    • 2006
  • Currently, boars selected for commercial use as AI sires are evaluated on grow-finish performance and carcass characteristics. If AI sires were also evaluated and selected on semen production, it may be possible to reduce the number of boars required to service sows, thereby improving the productivity and profitability of the boar stud. The objective of this study was to estimate genetic correlations between production and semen traits in the boar: average daily gain (ADG), backfat thickness (BF) and muscle depth (MD) as production traits, and total sperm cells (TSC), total concentration (TC), volume collected (SV), number of extended doses (ND), and acceptance rate of ejaculates (AR) as semen traits. Semen collection records and performance data for 843 boars and two generations of pedigree data were provided by Smithfield Premium Genetics. Backfat thickness and MD were measured by real-time ultrasound. Genetic parameters were estimated from five four-trait and one five-trait animal models using MTDFREML. Average heritability estimates were 0.39 for ADG, 0.32 for BF, 0.15 for MD, and repeatability estimates were 0.38 for SV, 0.37 for TSC, 0.09 for TC, 0.39 for ND, and 0.16 for AR. Semen traits showed a strong negative genetic correlation with MD and positive genetic correlation with BF. Genetic correlations between semen traits and ADG were low. Therefore, current AI boar selection practices may be having a detrimental effect on semen production.