• Title/Summary/Keyword: $\textrm{NO}_3$-N

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Characterization of Humanized Antibody Produced by Apoptosis-Resistant CHO Cells under Sodium Butyrate-Induced Condition

  • Kim, No-Soo;Chang, Kern-Hee;Chung, Bo-Sup;Kim, Sung-Hyun;Kim, Jung-Hoe;Lee, Gyun-min
    • Journal of Microbiology and Biotechnology
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    • v.13 no.6
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    • pp.926-936
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    • 2003
  • Overexpression of human Bcl-2 protein in recombinant Chinese hamster ovary (rCHO) cells producing humanized antibody (SH2-0.32) considerably suppressed sodium butyrate (NaBu)-induced apoptosis during batch culture by using commercially available serum-free medium, which extended the culture longevity. Due to the extended culture longevity provided by the anti-apoptotic effect of Bcl-2 overexpression, the final antibody concentration of 14C6-bcl-2 culture (Bcl-2 high producer, $23\;\mu\textrm{g}\;ml^{-1}$) was 2 times higher than that of the $SH2-0.32-{\Delta}bcl-2$ culture (cells transfected with bcl-2-deficient plasmid, $10.5\;\mu\textrm{g}\;ml^{-1}$) in the presence of NaBu. To determine the effect of NaBu/Bcl-2 overexpression on the molecular integrity of protein products, antibodies purified from 14C6-bcl-2 and $SH2-0.32-{\Delta}bcl-2$ cultures in the presence of NaBu were characterized by using various molecular assay systems. For comparison, antibody purified from the parental rCHO cell culture (SH2-0.32) in the absence of NaBu was also characterized. No significant changes in molecular weight of antibodies could be observed by SDS-PAGE. From GlycoSep-N column analysis, it was found that the core oligosaccharide structure ($GlcNAc_2Man_3GlcNAc_2$) was not affected by NaBu/Bcl-2 overexpression, while the microheterogeneity of N-linked oligosaccharide structure was slightly affected. Compared with the antibody produced in the absence of NaBu, the proportion of neutral oligosaccharides was increased from 10% (14C6-bcl-2) to 16% ($SH2-0.32-{\Delta}bcl-2$) in the presence of NaBu, which was accompanied by the reduced proportion of acidic oligosaccharides, especially of monosialylated and disialylated forms. The changes in microheterogeneous oligoformal structures of antibody in turn affected the mobility of antibody isoforms in isoelectric focusing (IEF), resulting in the occurrence of some more basic antibody isoforms produced in the presence of NaBu. However, the antigen-antibody binding properties were not changed by alteration of glycosylation pattern. The competitive enzyme-linked immunosorbent assay (ELISA) showed that the antibody produced by NaBu/Bcl-2 overexpression maintained its antigen-antibody binding properties with binding affinity of about $2.5{\times}10^9{\;}M^{-1}$. Taken together, no significant effects of NaBu/Bcl-2 overexpression on the molecular integrity of antibodies, produced by using serum-free medium, could be observed by the molecular assay systems.

Neuronal Dysfunction in Patients with Chronic Alcoholism Evaluated by In Vivo $^1H$ Magnetic Resonance Spectroscopy (알콜중독환자의 신경기능 장애: 생체 양성자 자기공명분광 연구)

  • Bo-Young Choe;Euy-Neyng Kim;Chang-Wook Lee;In-Ho Baik;Kwang-Soo Lee;Byung-Chul Son;Heung-Jae Chun;Hyoung-Koo Lee;Tae-Suk Suh
    • Investigative Magnetic Resonance Imaging
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    • v.4 no.2
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    • pp.94-99
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    • 2000
  • Purpose : With the use of localized, water-suppressed in vivo $^1H$ magnetic resonance spectroscopy (MRS), we evaluated the proton metabolic alterations in patients with chronic alcoholism and healthy normal controls. Material and Methods : Patients with chronic alcoholism (N = 10) and normal control subjects (N = 10) underwent MRS examinations using a stimulated echo acquisition mode (STEAM) pulse sequence with $2{\times}2{\times}2{\;}\textrm{cm}^3$ volume of interest (VOI) in the left cerebellum and basal ganglia. Proton metabolite ratios relative to creative (Cr) were obtained using a Marquart algorithm. Results : The specific feature in patients with chronic alcoholism was a significant decrease of N-acetylaspartate (NAA)/Cr ratio in the left cerebellum, compared with normal controls. No clear correlation of other metabolite ratios such as choline (Cho)/Cr and inositols (Ins)/Cr was established. Conclusion : Our preliminary study suggests that the reduction of NAA/Cr ratio may indicate neuronal loss in patients with chronic alcoholism. Thus, in vivo 1H MRS may be a useful modality in the clinical evaluation of patients with chronic alcoholism based on the proton metabolite ratios.

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Mutagenicity of Typhoid Vaccine

  • Li, Guang-Xun;Kang, Byeong-Cheol;Lee, Won-Woo;Ihm, Jong-Hee;Jung, Ji-Youn;Lee, Yong-Soon
    • Toxicological Research
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    • v.15 no.1
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    • pp.75-78
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    • 1999
  • In order to evaluate the mutagenic potential of Typhoid vaccine, 3 sets of mutagenicity tests were performed. In the reverse mutation test using Salmonella typhimurium TA98, TA100, TA1535 and TA1537, Typhoid vaccine did not increase the number of revertant at the doses of 100, 50, 25, 12.5, 6.25 $\mu\textrm{g}$/plate. I n chromosome aberration analysis using CHO cells were not found chromosomal aberration in different concentrations with or without metabolic activation at the doses of 0.25 mg/ml, 0.5mg/ml, 1mg/ml. In mouse micronucleus test, no significant increase in the occurrence of micronucleated polychromatic erythrocytes was observed in ICE male mice intramuscularly administered with Typhoid vaccine at the dosed of 0.1 mg/ml, 0.5 mg/ml, 1mg/ml. These results indicate that Typhoid vaccine gas no mutagenic potential in these in vitro and in vivo systems.

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Studies on Oocyte Activation Regimen for Nuclear Transfer in Hanwoo(Korean Cattle) (핵이식을 위한 한우 난자 활성화 처리방법에 관한 연구)

  • 임기순;양보석;박성재;양병철;장원경;박창식
    • Korean Journal of Animal Reproduction
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    • v.24 no.3
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    • pp.281-288
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    • 2000
  • This experiment was carried out to investigate the optimal activation condition for parthenogenetic development. In order to activate oocytes at 22 h post onset of maturation, the oocytes were subjected to 5 $\mu$M ionomycin(I) for 5 min ,10 $\mu$M calcium ionophore(Ca) for 5 min, 2 mM 6-dimethylamino-purine(DMAP) for 3 h and 10 $\mu\textrm{g}$/$m\ell$ cycloheximide(CH) for 6 h alone or in combination. The activated oocytes were cultured in modified CR$_1$aa at 5% $CO_2$, 5% $O_2$, 90% $N_2$. l. The cleavage rates after 48 h culture of oocytes treated with I, Ca, DMAP and CH were 12.7%, 14.1%, 28.9% and 22.9%, respectively. There was no blastocyst formation. 2. The cleavage rates after 48 h culture of oocytes treated with I + DMAP, I + CH, Ca + DMAP and Ca + CH were 96.9%, 82.1%, 93.1% and 34.7%, respectively. Developmental rates to blastocysts were 10.4%, 5.3%, 17.6% and 7.1 %, respectively. When oocytes were treated with Ior Ca followed by DMAP, the blastocyst formation rate was significantly higher than other groups(P <0.05). 3. According to single activation treatment, pronucleus formation rates were 5.4%, 3.6%, 28.3% and 28.8%, respectively, Whereas, all oocytes treated with the combined activation agents formed 100% pronucleus.

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Growth Characteristics of Creeping bentgrass Cultivars (크리핑 벤트그래스 품종의 생육 특성)

  • 이혜원;정대영;심상렬
    • Asian Journal of Turfgrass Science
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    • v.17 no.2_3
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    • pp.87-97
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    • 2003
  • The growth characteristics of creeping bentgrass seeded in the seaside landfill golf course are as follows. 1. As a result of analysis on the chemical characteristics of soil mixture used for turfgrass ground in this experiment, soil pH was 7.9, which is somewhat higher than the creeping bentgrass reference value of pH 5.5∼6.5; T-N(%) was 0.02, which is proper for the reference value, and trace element too lacked. 2. The cultivar with the fastest water infiltration was Seaside II recorded as 226.2cm/sec, while that with the slowest water infiltration was Pennlinks recorded as 141.1 cm/hr which was too faster than the USGA water infiltration reference value of 15∼30cm/hr. For the surface hardness of turfgrass ground with different cultivars, no statistically significant variation was found between the Penncross grass and the Pennlinks recorded as 18.6mm and 19.1 mm, respectively. The soil penetration was the highest in Pennlinks recorded as 7.6kg/$\textrm{cm}^2$ and lowest in Penn A-1 as 6.1kg/$\textrm{cm}^2$. 3. As a result of evaluation on visual quality at the early stage of growth, Penncross showed the most excellent visual quality compared to the others. However, Penn A-1 showed the most excellent visual quality at a late stage of growth around Sep. 17, 2003, and it was also excellent in the evaluation of visual color. Seaside II showed higher density around the root and the longest root length and was highly resistant to salt compared to others, but the initial sprouting rate was not satisfied, and the visual quality in the summer season was inferior to others. 4. As a result of measurement of the traffic injury, Penncross showed the most tolerant to the traffic stress and Pennlinks showed the most susceptible.

Study on the Physical Property of Stretch Film for Wrapping of Roughage (조사료 래핑용 스트레치 필름의 물리적 특성에 관한 연구)

  • 이성현;장유섭;박원규;최광재;김종근
    • Journal of Animal Environmental Science
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    • v.5 no.2
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    • pp.79-86
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    • 1999
  • Recently more dairy farmers are interested in the utilization of stretch film for wrapping silage material. Most of stretch film are imported from abroad and one domestic product was developed, but its quality has not yet been tested. Therefore this study was executed to offer basic data on this material to the farmers. Measured items were tensile load, extension ratio, and tear propagation strength of stretch films. Comparisons were made among the white color of domestic product(Kw), and white(Fw), black (Fb) and green color(Fg) of foreign products. The result were summarized as follows: 1. Tensile loads of lengthwise direction in stretch films were 557.4, 377.4, 282.6 and 398.4kgf/$\textrm{cm}^2$ in Kw, Fw, Fb and Fg, respectively, and tensile loads of width direction were 415.9, 418.1, 360.2 and 433.0kgf/$\textrm{cm}^2$ in Kw, Fw, Fb and Fg, respectively. 2. Extension ratios of lengthwise direction were 650, 462.5, 512.5% and 537.5% in Kw, Fw, Fb and Fg, respectively, Extension ratios of width direction were 930, 962.5, 950 and 1,000% in Kw, Fw, Fb and Fg, respectively. 3 Tear propagation strength of lengthwise direction 187.9, 148.9, 157.3 and 142.8kgf/cm, and tear propagation strength of width direction were 141.4, 129.8, 140.4 and 106.6kgf/cm in Kw, Fw, Fb and Fg, respectively. 4. In the light transmittance, there was no difference between white and green color stretch film, but it was very low in black one. Therefore, it is thought that research on quality of roll bale silage treated with each stretch film would be needed.

Long-Term Effects of the DHA Supplementation on Physical and Brain Development in Full-Term Infants (장기간에 걸친 DHA 보충이 영아의 신체발육 및 두뇌발달에 미치는 영향)

  • 정현주
    • Journal of Nutrition and Health
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    • v.31 no.8
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    • pp.1295-1306
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    • 1998
  • Recent research indicates that the n-3 fatty acid , docosahexaenoic acid(22 : 6n 3, DHA) plays an essential role in infant brain development . DHA is highly concentrated in brain and retinal tissues and accumulates during late fetal and early neonatal life. Diets deficient in DHA are associated with reduced levels of DHA in brain and retinal tissues. The purpose of this study is to investigate the long term effects of DHA supplementation on the growth and mental development of full-term infants. THirty four healty infants were recruited from those who were delivered at Kyung Hee Medical Center. The experimental groups were the breast milk+DHA(-) group who were fed human milk for 20 weeks after birth and thereafter were fed placebo formula for 28 weeks, the breast milk+DHA(+) group who were fed human milk for 20 weeks after birth and thereafter were fed DHA supplemented formula for 28 weeks, DHA(-) group who were fed placebo formula for 48 weeks, and DHA(+) group who were fed DHA supplemented formula for 48 weeks. The daily average intake of DHA for the breast milk+DHA(-) , breast milk+DHA(+), DHA(-) and DHA(+) groups were 39.1mg, 89.9mg, 17.7mg, and 160.224mg, respectively. The results showed that measurements of infant weight, length, head, and chest circumferncewere all in normal range and they were not influenced by the DHA supplements in their diets. There was a significant correlation between dietary DHA intake and erythrocyte DHA level. The results of flash visual evoke potential (VEP) test were not correlated with eerythrocyte DHA and dietary DHA levels at 48 weeks of age. No differences were found in Bayley mental and Psychomotor Development lndex scores among the four experimental groups at 48 weeks of age. Unlike the short-term effects there was no long-term effect of relatively small amounts of dietary DHA supplements on the scores for flash VEP and Bayley test, even thour호 there was an elevated DHA supplements on the scores for flash VEP and Bayley test, even through there was an elevated DHA content in the infants erythrocytes.

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Antimutagenic and Antioxidative Effects of Methanol Extract of Pine Pollen (송화 메탄올 추출물의 항산화적 항돌연변이 효과)

  • 박정섭;안병용;최동성
    • The Korean Journal of Food And Nutrition
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    • v.16 no.4
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    • pp.303-309
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    • 2003
  • This study was performed to investigate the antimutagenic and antioxidative activities of pine pollen with respect to the microbial mutation induced by various mutagens such as 1-NP, daunomycin, 2-NF, MNNG, NaN$_3$, 4NQO, 4-NOPD, AFB$_1$, Trp-P-1, 2-AF and oxidative mutagens such as t-BOOH, H$_2$O$_2$. Pine pollen, originally extracted with hexane, was reextracted with 70% methanol. The results obtained using the methanol extract, in terms of the antimutagenicity observed in relation to ten kinds of mutagens, showed that it exhibited 17.8, 82.2 and 80.9% inhibitory effects against daunomycin, AFB$_1$, and Trp-P-1, respectively, in Salmonella. typhimurium TA98 and a 72.3% inhibitory effect against AFB$_1$in S. tyPhimurium TA100. In terms of the antimutagenicity exhibited in relation to t-BOOH, a 72.3% inhibitory effect was observed, but no antimutagenicity was observed in relation to the other mutagens and strains. The methanol extract was further fractionated by chloroform, ethyl acetate, n-butanol. In S. typhimurium TA98, the chloroform(150 $\mu\textrm{g}$/plate) fraction showed strong antimutagenic effects of 55.6%, 93.7% and 93.5%, while the ethyl acetate(100 $\mu\textrm{g}$/plate) fraction showed 11.4%, 74.3% and 85.2% in relation to the mutagenicity induced by daunomycin, AFB$_1$and Trp-P-1, respectively. In S. typhimurium TA100, the chloroform and ethyl acetate fractions showed antimutagenic effects of 95.1% and 62.5%, respectively, on the mutagenicity induced by AFB$_1$. In S. typhimurium TA102, the chloroform fraction showed an antimutagenic effect of 93.6% on the mutagenicity induced by t-BOOH.

Properties of $Sr_{0.8}Bi_{2.3}{(Ta_{1-x}Nb_{x})}_{2}O_{9+{\alpha}}$ Thin Films

  • Park, Sang-Jun;Jang, Gun-Eik
    • Transactions on Electrical and Electronic Materials
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    • v.1 no.1
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    • pp.22-25
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    • 2000
  • Polycrystalline SBTN layered ferroelectric thin film with various Nb mole ratios were prepared by sol-gel method Pt/ $SiO_2$/Si (100) substrates. The films were annealed at different temperature and characterized in terms of phase and microstructure. The films were crystallized with a high (105) diffraction intensity and had rodike structure, SBTN films fired at 800$^{circ}C$ revealed standard hysteresis loops with no fatigue for up to 10$^{10}$ cycles. At an applied voltage of 5V the dielectric constant($varepsilon$) , dissipation factor (tan $delta$), remanent polarization(ZPr) and coercive field(Ec) of typical S $r_{0.8}$B $i_{2.3}$(T $a_{1-x}$ N $b_{x}$) $O_{9+}$$alpha$/ thin film(x=0.1) prepared on Pt/ $SiO_2$/Si (100) were about 277.7, 0.042, 3.74$mu$C/$textrm{cm}^2$, and 24.8kv/cm respectively.ly.y. respectively.ly.y.y..

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In Vitro Production of Bovine Embryos by Modification of Simple Defined Culture Medium (단순한정배양액의 성분조정에 의한 소 수정란의 체외생산)

  • 노상호;윤종택;한기영;이병천;황우석
    • Journal of Embryo Transfer
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    • v.13 no.3
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    • pp.235-243
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    • 1998
  • In this study, we investigated the effects of three kinds of culture medium (Charles and Rosenkrans; CRlaa, Tyrode's; TALP, synthetic oviduct fluid: SOF), insulin transferrin + selenium complex (ITS), macromolecules(polyvinyl alcohol: PVA, fetalb-ovine serum: FBS) and NaCl on the development of early bovine embryos. In experiment 1, there were no differences in embryo development among three kinds of embryo culture medium (CR $l_{aa}$ , TALP, SOF). In experiment 2, BSA, FBS and PVA were added each in TALP as macromolecule sources. The developmental rates of embryos in BSA or FBS added TALP were significantly higher than in PVA added one (p〈0.01), but there was no difference between BSA and FBS added groups. In experiment 3, bovine embryos were cultured in TALP with the following supplements: BSA alone(1, 3 or 8 mg/ml, each) or BSA(1, 3 or 8 mg/ml, each)+ITS (10$\mu\textrm{g}$/m1 insulin, 5 $\mu\textrm{g}$/ml transferrin, 5 ng/ml selenium). In higher concentration of BSA and ITS supplemented groups, the developmental rates over compacted morula were higher than others, but there was a significant effect of ITS only in 1 mg/ml of BSA added group (p〈0.05). In experiment 4, the effect of reduced concentration of NaCl was evaluated. The developmental rate over compacted morula in the medium containing 90 mM of NaCl was higher than in 114 mM group (p〈0.05). In conclusion, BSA could be used as a macromolecule source in bovine embryo culture, and ITS, as a serum substitute, could be used for improving of embryonic development. Also, reduction of NaCl concentration from 114 mM to 90 mM may improve the development of bovine embryos.bryos.

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