• Title/Summary/Keyword: two-stage culture

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SUPPRESSION OF PHORBOL ESTER-INDUCED EXPRESSION OF CYCLLOOXYGENASE-2 AND INDUCIBLE NITRIC OXIDE SYNTHASE BY SELCTED CHEMOPREVENTIVE PHYTOCHEMICALS VIA DOWN-REGULATION OF NF-$\textsc{k}$B

  • Surh, Young-Joon
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05b
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    • pp.88.2-98
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    • 2002
  • A wide arry of naturally occurring substances particularly those present in dietary and medicinal plants, have been reported to possess substantial cancer chemopreventive properties. Certain phytochemicals retain strong antioxidative and anti-inflammatory properties which appear to contribute to their chemopreventive or chemoprotective activities. Inducible cyclooxygenase(COX-2) and nitric oxide synthase (iNOS) are important enzymes that mediate inflammatory processes. There is some evidence that expression of both COX-2 and iNOS is co-regulated by the eukaryotic transcription factor NF-$textsc{k}$B. Increased expression of COX-2 and/or iNOS has been associated with pathophysiology of certain types of human cancers as well as inflammatory diseases. Since inflammation is closely linked to tumor promotion, substances with potent anti-inflammatory activies are anticipated to exert chemopreventive effects on carcinogenesis, particularly in the promotion stage. An example is curcumin, a yellow pigment of turmeric (Curcuma longa L., Zingiberaceae), that strongly occurring diaryl heptanoids structurally related to curcumin have substantial anti-tumor promotional activities in two-stage mouse skin carcinogenesis. Thus, yakuchinone A [1-(4'-hydroxy-3'-methoxyphenyl)-7-phenyl-3heptanone] and yakuchinone B [1-(4'-hydroxy-3'methoxyphenyl)-7-phenylhept-1-en-3-one] present in Alpinia oxyphylla Miquel (Zingiberacease) attenuate phorbol ester-induced inflammation and papilloma formation in female ICR mice. These diarylheptanoids also suppressed phorbol ester-induced activation of epdermal ornithine decarboxylase and its mRNA expression when applied onto shaven backs of mice. Yakuchinone A and B as well as curcumin inhibited phorbol ester-induced expression of COX-2 and iNOS and their mRNA in mouse skin via inactivation of NF-$textsc{k}$B. Capsaicin, a major pungent ingredient of red pepper also attenuated phorbol ester-induced NF-$textsc{k}$B activation. Similar suppression of COX-2 and iNOS and down-regulation of NF-$textsc{k}$B activation for its DNA binding were observed with the ginsenosied Rg3 and the ethanol extract of Artemisia asiatica. We have also found that certain anti-inflammatory phytochemicals exert inhibitory effects on phorbol ester-induced COX-2 expression and NF-$textsc{k}$B activation in immortalized human breast epithelial (MCF-10A) cells in culture. One of the plausible mechanisms undelying inhibition by aforementioned phytochemicals of phorbol ester-induced NF-$textsc{k}$B activation involves interference with degragation of the inhibitory unit, I$textsc{k}$Ba, which blocks subsequent nuclear translocation of the functionally active p65 subunit of NF-$textsc{k}$B. the activation of epidermal NF-$textsc{k}$B by phorbol ester and subsequent induction of COX-2 hence appear to play an important role in intracellular signaling pathwasy leading to tumor promotion and targeted inhibition of NF-$textsc{k}$B may provide a new promising cancer chemopreventive strategy.

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Diversity of Uncultured Microorganisms Associated with the Anaerobic Pentachlorophenol Degradation Estimated by Comparative RELP Analysis of PCR-Amplified 16S rDNA Clones (16S rDNA 클론들의 RFLP 비교분석에서 얻어진 Pentachlorophenol의 혐기성 분해에 따른 미생물군집의 변화)

  • 성창수;권오섭;박영식
    • Korean Journal of Microbiology
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    • v.33 no.2
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    • pp.149-156
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    • 1997
  • We have employed comparative RFL,P(Restriction Fragment Ixngth Pol~iniorphism) analysis and molecular phylogenetic techniques to investigate the diversity of uncultured microorganisms associated with the anaerobic PCP degradation in PCP-adapted enrichment cultures inoculated by samples from anaerobic cewage sludgc(Jangrim, Pusan) and leachate of landfill site(Kimhae). 16s rDNA cloncs were obtairted by PCR amplification of mixed population DNAs extracted directly from the nonactive and active stage ol each PCP-adapted culture. After three rounds of comparative RFLP analyses. two RFLP types. designated as Ala and Hld, were found prevalent and common in both active stage samples. Thc analysis of phylogenctic diversity bawd on the 5'-terminal 180 nt of sequences from whole clones of the Ala and Bld RFLP types showed close similarity among themselves. In case of Bld clones, 7XQ of them shared identical sequences. Thcse resuliq suggest that the clones of both RFLP types wcre originated from highly affiliated microorganisms which are e~iriched as a result of metabolic activity to PCP. The full-length 16s rRNA sequence of each representative clone from both RFLP types was determined. and an Ala clone w i n found to he related to Clo.strrdiurn ulfutzac~(Genk~ank No. Z69203) and a Bld clone to Thermobacteroides proteolyticus(Genbank No. X09335), with sequence similarities of 89%' and 97%. respectively.

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Comparing Malaysian Youths and Adults'toward Hallyu Consciousness (말레이시아 청소년과 성인들의 한류 의식 비교)

  • Cho, Kum-Ju;Jang, Won-Ho
    • The Journal of the Korea Contents Association
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    • v.13 no.9
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    • pp.92-101
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    • 2013
  • Hallyu in Malaysia is considered to be in the growth stage as in countries such as Philippine, Myanmar, Hungary, and Uzbekistan. In the growth stage, Hallyu becomes increasingly popular and familiar to the general public and the sales of Hallyu-related products starts to increase. The purpose of this research is to compare how Hallyu is perceived among Malaysian youths and adults. Analysis is carried out using FGD. The two groups are asked about what they think of Korea and the success of Hallyu, how much interest they have in foreign cultures in general, and how they come into contact with them. Based on these results, problems currently facing Hallyu and possible solutions are examined. The analysis shows that the interest in Hallyu among Malaysians varies according to age and gender. Both male and female participants in their 20's have shown a much greater interest in Hallyu than the participants over 30's. But Malaysians are known to be relatively open-minded about accepting foreign cultures, and this is reflected in the current study. They view the popularity of Korean drama, movies, and music as a trend, rather than being intolerant of them. There is no concern for negativity toward Hallyu in Malaysia, but it is likely that Hallyu may be replaced by another foreign culture unless its influence is constantly maintained in the area. Therefore, it is essential to develop measures and models to not only retain, but strengthen Hallyu's influence.

Characteristics of microorganism isolated from Cotton Waste Media for the Oyster Mushroom Cultivation (느타리버섯 균상재배 중 배지에서 분리한 미생물의 특성)

  • Lee, Chan-Jung;Jhune, Chang-Sung;Cheong, Jong-Chun;Oh, Jin-A;Han, Hye-Su;Um, Na-Na
    • The Korean Journal of Mycology
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    • v.38 no.2
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    • pp.120-124
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    • 2010
  • This study was carried out to investigate interaction between mushroom mycelium and microorganisms in cotton waste media for the shelf cultivation of oyster mushroom. Two oyster mushroom farms was selected for this experiment. One was good mushroom farm (farmhouse I) and the other failed mushroom farm (farmhouse II). In farmhouse I, the inhibition microorganisms were higher toward the end of growth stage than the early stage, but the result of farmhouse II was opposite. Effects of the mycelium growth on plate culture showed same results on mushrooms as the earlier one. And the mycelium growth was influenced by secretory materials of microorganisms. Among of the isolates, Only few microorganism had inhibitory effects on either P. tolaasii or T. harzianum causing the disease of oyster mushrooms. But more microorganisms had inhibition effects on P. agarici.

Development of Saccharomyces cerevisiae Strains with High RNA Content (리보핵산을 다량으로 함유하는 Saccharomyces cerevisiae 균주의 개발)

  • Kim, Jae-Sik;Kim, Jin-Wook;Shim, Won;Min, Byoung-Cheol;Kim, Jung-Wan;Park, Kwan-Hwa;Pek, Un-Hua
    • Korean Journal of Food Science and Technology
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    • v.31 no.2
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    • pp.465-474
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    • 1999
  • RNase activity of Saccharomyces cerevisiae ATCC 7754 was investigated to obtain strains with high ribonucleic acid (RNA) content. The yeast strain contained two RNase activities; an acidic RNase with a optima of pH $3{\sim}4$ and an alkaline RNase with a optima pH 9. The acidic RNase activity was inhibited by $0.08\;M\;HgCl_{2}$ most drastically. The alkaline RNase activity was inhibited by 2.0 M NaCl or KCl, while enhanced by addition of $0.05\;M\;CaCl_{2},\;0.02\;M\;ZnSO_{4},\;or\;0.008\;M\;HgCl_{2}$. Various mutants of Saccharomyces cerevisiae ATCC 7754 were isolated by ethylmethane sulfonate (EMS) treatment or $\gamma$-ray/ultra violet irradiation. Among the mutants that were sensitive to high concentration of KCl which inhibits alkaline RNase, B24 was selected for high RNA content per culture volume. Growth characteristics of the mutant were comparable to those of the mother strain with optimum growth at pH $4.5{\sim}5.5$. The mutant accumulated higher content of RNA than the mother strain when glucose was used as the carbon source. However, both growth rate and total RNA content of the mutant were higher in molasses medium than in glucose medium. RNA content of the mutant increased rapidly during the early stage of growth, and then decreased gradually until the culture reached stationary phase by a fed-batch culture in a 5 L jar fermenter. Maximal cell harvest and the final RNA content using the mutant B24 were 69.6 g/L culture broth and 19.8 g/100 g of the dry cell while those using the mother strain were 68 g/L culture broth and 16.1 g/100 g of dry cell, respectively.

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The Effect of EDTA and Fetal Cord Serum Supplementation to Ham's F-10 Culture Medium on Developmental Potential of Mouse Embryos In Vitro (Ham's F-10 배양액에 첨가된 태아제대혈청과 EDTA가 백서 수정란의 분할에 미치는 영향)

  • Kim, Byeong-Seog;Lee, Young-Gi;Park, Yoon-Kee;Lee, Tae-Hyung;Lee, Sung-Ho
    • Journal of Yeungnam Medical Science
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    • v.12 no.1
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    • pp.124-134
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    • 1995
  • It is the most important to select optimal culture conditions to promote safe embryo growth in the technique of human in vitro fertilization and embryo transfer. It has been shown that the addition of biologic fluids, such as blood serum, of various origins, improved fertilization and early cleavage rates in numerous species. The purpose of this study is to attempt to measure developmental potential of mouse eggs fertilized and cleaved in Ham's F10 culture medium containing a chelating agent, EDTA and fetal cord serum. In this study, we selected 40 female mice and 20 male mice, and investigated optimal serum concentrations for mouse embryo growth. Two cell stage mouse embryos were cultured in Ham's F-10 medium, Ham's F-10 medium with various concentrations of EDTA, or Ham's F-10 medium with EDTA and 10% human cord serum. Developmental ratios to morula in Ham's F-10 medium containing various concentrations of EDTA and/or 10% fetal cord serum were significantly higher than in unsupplemented Ham's F-10 medium (p<0.05). Developmental ratios to blastocyst in Ham's F-10 containing 10% fetal cord serum and $50{\mu}M$ or $100{\mu}M$ EDTA were significanltly higher than in unsupplemented Ham's F-10 medium (p<0.05). Developmental ratios to morula in Ham's F-10 containing 10% fetal cord serum and $100{\mu}M$ EDTA were significanltly higher than in Ham's F-10 with 10% fetal cord serum used commonly in many human IVF centers(p<0.05). Developmental ratio to blastocyst in Ham's F-10 containing 10% fetal cord serum and $100{\mu}M$ EDTA was significanlty higher than in Ham's F-10 with $200{\mu}M$ EDTA(P<0.05). In summary, embryo development to morula and blastocyst was significanlty higher in the presence of human cord serum or EDTA than in the unsupplemented medium. The most significanly development to morula and blastocyst was obtained at Ham's F-10 medium with $100{\mu}M$ concentration of EDTA and 10% fetal cord serum. These results suggest that Ham's F-10 medium containing 10% fetal cord serum and optimal concentrations of EDTA significantly promoted early cleavage of mouse zygotes, and these will be useful as basic data for the selection of culture medium in human in vitro fertilization.

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Cencept of adaptability for heavy nitrogen in view of nitrogen metabolism -II. Nitrogen metabolism under the change of itrogen nenvironment (내비성(耐肥性)에 관(關)한 질소대사적(窒素代謝的) 개념(槪念) -II. 질소환경(窒素環境) 변화(變化)에 따른 질소대사(窒素代謝))

  • Yoon, Jong Hyuk;Park, Hoon;Cho, Sung Jin
    • Korean Journal of Soil Science and Fertilizer
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    • v.7 no.1
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    • pp.49-53
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    • 1974
  • Varietal difference in nitrogen metabolism was investigated under water culture system with high (50 or 80ppm) and low (10 or 40 ppm) levels of nitrogen and with two-week minus nitrogen treatment at maximum tillering and heading stage using a leading local variety, Jinheung and high-yielding IR667 line (newly bred tropical variety). 1. In high nitrogen level Jinheung showed higher yield than IR667. vise versa in low nitrogen level. 2. Poor yield of IR667 at high nitrogen may be due to ammonium toxicity that was eliminated by minus nitrogen from culture solution. 3. IR667 was more sensitive to the change of nitrogen environment. 4. With high nitrogen medium Jinheung showed nigher nitrogen uptake and higher capacity of protein synthesis than IR667, and vise versa at low nitrogen medium. 5. From the above facts it could be concluded that Jinheung has higher metabolic adaptability for heavy nitrogen while IR667 has higher structural adaptability for heavy mitrogen and that better productive adaptability will be resulted in the combination of both characteristics.

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In vitro Fertilization and Development of Pig Oocytes Inseminated with Boar Sperm by Different Sperm Washing Media after Thawing of the Frozen Straws

  • Yi, Y.J.;Ko, H.J.;Lee, S.H.;Yang, C.B.;Son, D.S.;Kim, H.K.;Park, C.S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.2
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    • pp.164-167
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    • 2004
  • This study was carried out to investigate in vitro fertilization and development of in vitro matured pig oocytes inseminated with the Duroc boar sperm by different sperm washing media after thawing of the 5 ml frozen straws. Immature follicular oocytes (30-40) were transferred into each well of a Nunc 4-well multidish containing $500{\mu}l$ mTCM199 maturation medium. The sperm rich portion of ejaculates was collected into a 250 ml insulated vacuum bottle and gradually cooled 22 to $24^{\circ}C$ over a 2 h period. Semen was centrifuged at 800 g for 10 min and the seminal plasma discarded. Sperm were esuspended in a lactose-egg yolk and N-acetyl-Dglucosamine (LEN) diluent to contain $1{\times}10^{9}$ sperm/ml and cooled to $5^{\circ}C$ over a 2 h period. Immediately before freezing, semen was rediluted with an equal volume of LEN+4% glycerol and packed into 5 ml straws. After thawing of the 5 ml straw, the 5 ml semen was diluted with 20 ml Beltsville thawing solution (BTS) at room temperature. Oocytes were inseminated with untreated (unwashed and nonpreincubated) or treated sperm (washed two times in BTS, mTLP-PVA and mTBM media, respectively and nonpreincubated) with $2{\times}10^{7}$ sperm concentration. Oocytes were coincubated for 6 h in $500{\mu}l$ mTBM fertilization. At 6 h after IVF, oocytes were transferred into $500{\mu}l$ NCSU-23 culture medium for further culture of 6 h. Sperm penetration, polyspermy and male pronuclear formation of oocytes at 12 h after IVF and developmental ability of oocytes at 48 h after IVF were evaluated. Sperm penetration rate, male pronuclear formation and rate of cleaved embryos were higher in the BTS, mTLP-PVA and mTBM treatments than the unwashed treatment (p<0.05). The rate of blastocysts from the cleaved oocytes (2-4 cell stage) were higher in the mTLP-PVA treatment than in the unwashed, BTS and mTBM treatments. In conclusion, we recommend the washing of frozen-thawed sperm with mTLP-PVA medium before in vitro fertilization of oocytes in mTBM medium.

Problems and Solutions of Zymography Techniques (자이모그라피 기술의 문제점과 해결)

  • Kang, Dae-Ook;Choi, Nack-Shick
    • Journal of Life Science
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    • v.29 no.12
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    • pp.1408-1414
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    • 2019
  • Enzymes are widely used in industrial applications such as detergents, food, feed production, pharmaceuticals and medical applications and major contributors to clean industrial products and processes. To screen, identify, and characterize the enzymes the zymography techniques are routinely used. The zymography technique is a simple, sensitive, and quantifiable technique that is widely used to detect functional enzymes following electrophoretic separation in sodium dodecyl sulfate (SDS)-polyacrylamide gels. The method is a versatile two-stage technique involving protein separation by electrophoresis followed by the detection of enzyme activity in polyacrylamide gels under non-reducing conditions. It is based on SDS-polyacrylamide gel (PAG) copolymerization with substrates, which are degraded by the hydrolytic enzymes restored in enzyme reaction buffer after the electrophoretic separation. Any kind of biological sample can be applied and analyzed on zymography, including culture supernatants of microbes, plants extracts, blood, tissue culture fluids, enzymes in foods extracts and metaproteome. The advantage of zymography is that it is possible to directly detect the protein with activity on the electrophoretic gel as well as confirm the activity at the nanogram level. Thus, this zymography technology can be applied in various fields. However, these advantages are rather disadvantageous and can often lead to experimental errors. In this review, the advantages, disadvantages, and problem solving of zymography technique are described.

Effect of Minimum Night Temperature on Growth of Seedlings of Pinus densiflora and Betula platyphylla in Container Culture during Winter Season (소나무, 자작나무의 겨울철 시설양묘시 생장초기 야간최저온도)

  • 홍성각;윤종규;윤택승;김종진
    • Journal of Bio-Environment Control
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    • v.11 no.4
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    • pp.163-167
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    • 2002
  • This study was carried out to investigate the effect of minimum night temperature in the PE house on growth of seedlings of Pinus densiflora and Betula platyphylla in winter season. The experiment was performed with three minimum night temperature regimes,5~8$^{\circ}C$, 10~13$^{\circ}C$ and 15~18$^{\circ}C$. The temperature regimes were maintained for 8 weeks (to April 6) after germination peak, and then were broken by increasing natural temperature. The temperature did not affect the germination rate but delayed the germination by 10 days. Height, root collar diameter growth, and dry weight of two species were reduced by relatively low night temperature. The reduction was more obvious in the dry weight of above the ground measured at 8 weeks after germination. The seedlings grown for 8 weeks at 5~8$^{\circ}C$ showed the lowest T/R ratio. It was also observed that the height growth response in Betula platyphylla seedlings to relatively low temperature was more sensitive than Pinus densiflora.