• Biotechnology and Bioprocess Engineering:BBE
• /
• v.6 no.2
• /
• pp.117-127
• /
• 2001

The high-level expression of a human single chain urokinase-type plasminogen activator (scu-PA) was achieved by employing a methotrexate (MTX)-dependent gene amplification system in Chinese hamster ovary (CHO) cells. By cotransfecting and coamplifying a scu-PA expression plasmid and dihydrofolate reductase (DHFR) minigene, several scu-PA expressing CHO cell lines were selected and gene-amplified. These recombinant cell lines, NGpUKs, secreted a completely processed scu-PA of 54 kD and up to 60mg/L was accumulated in the culture medium when they were adapted to an optimal MTX concentration. Over 95% of the scu-PA expressed was secreted in the culture medium and identified having the proper function of a plasminogen activator when activated by plasmin. Based on a genomic Southern analysis, a representative subclone, MGpUK-5, exhibited MTX-dependent scu-PA gene amplification, plus the initial single-copy gene of scu-PA eventually turned into about 150 copies of the amplified gene of scu-PA after gradual adaptation to 2.0$\mu$M of MTX. Meanwhile, the transcripts kof the scu-PA gene increased, although -early saturation of transcription was identified at 0.1$\mu$M of MTX. The scu-PA production by the MGpUK-5 subclone also increased relative to the gene amplification and increased transcripts, however, the relationship was not linearly proportional. Accordingly, since the MGpUK cell lines expressed elevated levels of enzymatically active scu-PA, these cell lines could be applied to the largescale production of scu-PA.

• Korean Journal of Soil Science and Fertilizer
• /
• v.6 no.4
• /
• pp.213-219
• /
• 1973

Effects of three slow-release nitrogen fertilizers (Sulfur-coated urea, (SCU) Jin Hae Chemical Co. Korea(JHC); Tennessee Valley Authority. U.S.A(TVA); METAP, Japan) on the yield and nitrogen uptake of IR667 (Suwon 213) were investigated comparing with urea split application under the field condition with early and usual saeson cultivation. 1. SCU(JHC) was not superior than urea split application in yield and nitrogen nutrition due to early release of nitrogen. 2. SCU(TVA) outyielded urea split by 10% showed always highest nitrogen content in straw and most balanced relative cumulative curve of nitrogen uptake or dry matter yield. 3. At ear formation stage SCU(TVA) showed specially high nitrogen content which was highly correlated with number of grain per $m^2$ of field. 4. Only SCU(TVA) showed one peak curve of nitrogen uptake rate at ear formation stage while others showed two peak pattern. 5. nitrogen fertilizer recovery of slow-release fertilizers was higher than urea split in early season cultivation and SCU (TVA) (46.0) was higher but SCU(JHC) (38.9) was similar in compare with urea split(37.1) in usual season cultivation. 6. Most promising slow-release nitrogen to match plant requirement seems to be one mixed of various slow-release fertilizers having different release pattern.

• KSBB Journal
• /
• v.9 no.1
• /
• pp.48-54
• /
• 1994

A modified amidolytic assay and a fibrin plate method were used to accurately measure the concentration of single chain urokinase type plasminogen activator (scu-PA) and two-chain urokinase type plasminogen activator (tc-PA) in the spent media. $1.65{\times}10^6$(viable cells/ml) of maximum cell density and 1670(IU/ml) of scu-PA concentration were obtained in 1% serum containing medium. The overall conversion ratio from scu-PA to tc-PA was less than 10%. In the results of batch cultivation in a spinner vessel, $4.43{\times}10^6(total cells/ml)$ of maximum cell density and 1560(IU/ml) of scu-PA concentration was observed. The maximum scu-PA concentration and specific scu-PA Productivity were obtained in 1760(IU/ml) and $3.13{\times}10^{-4}(IU/cell)$, respectively, from perfusion cultivation. The conveysion ratios from batch, fed-batch and perfusion cultivations were less than 12%, which means that about 90% of scu-PA secreted from the cells can be maintained during the cultivations.

• Journal of Bio-Environment Control
• /
• v.6 no.1
• /
• pp.1-14
• /
• 1997

This experiment was conducted to find out the compositions of nutrient solution for closed system in substrate culture of cucumber. Cucumber(Cucumis sativus L. cv. Eunsung baekdadagi) plants were grown in the substrates supplied with the nutrient solutions whose strengths were 1/2, 1, and 3/2 of the original concentration developed by National Horticultural Research Station in Japan. By increasing the nutrient concentrations, plant height decreased but leaf length, leaf width, and leaf number showed little differences. A number of marketable fruit and marketable yield were the highest in the concentration of 1 strength. The nutrient compositions of solution developed for closed system in cucumber substrate culture were N 11.4, P 3.3, K 6.0, Ca 4.5, and Mg 3.5 me.$\ell$$^{-1}$ during the vegetative growth period and N 10.4, P 3.3, K 5.0, Ca 4.5, and Mg 3,5 me.$\ell$$^{-1}$ during the reproductive growth period. To examine the suitability of nutrient solution developed in the above experiment, cucumber plants were grown in the substrates supplied with different solutions and concentrations - Yamasaki's nutrient solution(Yamasaki) of 1 S, nutrient solution of Research Station for Greenhouse Vegetable and Floriculture on the Netherlands(PTG) of 1 S, nutrient solution developed in the above experiment(SCU) of 1/2, 1, and 3/2 S. EC and pH in root zone changed little in the all treatments during growing period. As cucumber plants grew, the concentrations of N, P, and K in root zone decreased but Ca concentration increased. Net $CO_2$ assimilation rate of cucumber leaves was high in SCU of 1 and 3/2 S, and Yamasaki of 1 S. Growth of cucumber plants was the lowest in SCU of 1/2 S.

• Journal of Bio-Environment Control
• /
• v.6 no.4
• /
• pp.299-309
• /
• 1997

This experiment was conducted to determine the mineral absorption character and develop the optimum composition of nutrient solution for dill(Anethum graveolens L.) in hydroponics. Dill(Anethum graveolens L.) plants were grown in nutrient film technique(NFT) supplied with 1/4, 1/2, and 1 strength of the nutrient solution developed by National Research Station in Japan(HRS). Plants grown in 1/2 strength showed the best growth in plant height, fresh weight, and dry weight compared with those grown in 1/4 or 1 strength. In 1/2 strength solution, pH and EC changed little and proper nutrient contents were observed in the leaves as compared to plant nutrient diagnosis standard. Based on these results, optimum macronutrients were composed by nutrient- water absorption rate(n/w) with 1/2 strength: NO$_2$―N 8.85, NH$_4$―N 0.55, P 2.1, K 6.2, Ca 2.8, and Mg 1.7 me L$^{-1}$ To examine the suitability of the nutrient solution developed(SCU) , dill plants were grown in NFT supplied with two different kinds of solution and concentration. 1/2, 1, 3/2 and 25 of SCU and 1/2S of HRS. Changes of pH and EC were not distinct in 1S, but a significant change of pH was shown in low concentrations-HRS 1/2S and SCU 1/2S. Shoot fresh and dry weight were much higher in the plants grown in SCU IS as compared with HRS 1/2S. There were no significant differences in growth of plants grown in SCU IS, 3/2S, and 25. In addition, nutrient contents in the leaves grown by SCU 1S were in proper levels as compared with plant nutrient diagnosis standard. SCU 1S developed in this experiment was found to be optimum for dill in hydroponics.

• KSBB Journal
• /
• v.9 no.5
• /
• pp.518-524
• /
• 1994

Serum free medium was developed to cultivate Human Embryonic Kidney cells with Cytodex III microcarriers. The cells normally attached and spreaded on the microcarriers in serum free medium, and grew well by bead to bead processes. 85% of attachment yield was obtained on Cytodex III in this medium, compared to about 93% in 1% serum containing medium. About 90% of the attached HEK cells spreaded after 6 hours of post-attachment periods on the surface of microcarrier. Maximum cell density and scu-PA concentration in a serum free medium were $9.1{\times}10^5$ cells/ml and 1790 IU/ml, respectively, with fed-batch cultivation. Maximum cell density and scu-PA concentration in this medium with perfusion cultivation were $2.5{\times}10^6$ cells/ml and 1820 IU/ml, respectively. The conversion of single chain urokinase type plasminogen activator (scu-PA) into two chain type plasminogen activator (tc-PA) was less than 5% in a serum free medium compared to about 10% in 0.5% serum containing medium.

• Journal of Nutrition and Health
• /
• v.37 no.6
• /
• pp.431-439
• /
• 2004

Scutellariae Radix (Scu.), one of the immune-regulatory substances, is recognized to play the role in the metabolic process of inflammation, allergy and immunity. It has been traditionally used in the Oriental medicine to treat inflammatory bowel diseases (IBD). The purpose of this study was to evaluate the effects of water extracts of Scutellariae Radix on the spleen lymphocyte immune function in the Balb/c female mice treated with dextran sodium sulfate (DSS) to induce colitis. Water extract of Scutellariae Radix (100 mg/kg) and sulfasalazine (50 mg/kg) were administrated orally for 2 weeks of experimental period. Mice were divided into three experimental groups randomly: DSS group (5％ DSS was ad libitum for 5 days) as control group, DSS ＋ Scu. (water extracts of Scutellariae Radix for 2 weeks after 5％ DSS was ad libitum for 5 days) as experimental group, and DSS ＋ Sulfasalazine group (Sulfasalazine for 2 weeks after 5％ DSS was ad libitum for 5 days) as positive control group. Levels of Ig A, Ig E, CD4$^{＋}$, CD8$^{＋}$, TNF-$\alpha$ and other cytokines were measured. Treatment of DSS for 5 days induced bowel inflammation and the treatment with Scu. water exteract and sulfasalazine significantly recovered the damage. The length of intestine of DSS group was significantly shorter than that of other groups. The serum and fecal concentration of Ig A of SS ＋ Scu group was higher than those of DSS group. The contents of CD4$^{＋}$ T cells was higher in the DSS ＋ Scu. group than the other groups and CD8$^{＋}$ T cells was the lowest in DSS ＋ Sulfasalazine group. The Ig A level of cultured supernatant of spleen lymphocyte was the highest, while the Ig E level was the lowest in SS ＋ Scu group. The concentration of TNF-$\alpha$, cytokine secreted from the Th1 cell in the supernatant spleen lymphocyte, was the highest in the DSS group and the lowest in the DSS ＋ Scu. group. The concentration of IFN-${\gamma}$ and ll...-12 was lower in the DSS ＋ Scu. group than those of the other groups. The concentration of IL-4 in the supernatant of spleen lymphocyte was the lowest in the DSS ＋ Scu. group but IL-10 was not significantly different. Based on these findings, water extract of Scutellariae Radix exhibited the inhibitory effect via IL-4 production thereby inhibited the production of Ig E and strengthened immune system, and alleviated injury in DSS- induced colitis mice model.

• Proceedings of the Korea Concrete Institute Conference
• /
• 2002.05a
• /
• pp.817-822
• /
• 2002

In this paper, the strengthening analysis by strut-tie model for strengthened shear failed RC deep beam by the so-called the Steel Clamping Unit (SCU), which is a strengthening equipment, is carried out. The analysis considers the span-to-depth ratio, the existence of prestressing and stirrup, the shape of shear crack, and the strengthening position of the SCU. Based on analytical results, optimized strengthening analysis and design are carried out by investigating the behavior of the strengthened deep beams. The comparison between analytical results and experimental results shows that optimum strengthening effect by the SCU can be obtained when compressive strut zone created by SCU is away from major shear crack of the beam as far as possible.

• KSBB Journal
• /
• v.5 no.4
• /
• pp.391-396
• /
• 1990

4.0$\times$10-3 (IU/cells/day) of maximum specific scu-PA production, which was higher than those by a 75$\textrm{cm}^2$ T-flask and conventional perfusion systems, was maintained by tube-type bioreactor at 0.35 (1/h) of perfusion rate corresponding to 0.15 (dynes/$\textrm{cm}^2$) of shear stress. The production of scu-PA is also increased as shear stress was slowly increased, which is similar to natural human blood circulation. The tube reactor proves that there may be no limitation of oxygen supply by showing 1.0 (1/h) of oxygen transfer coefficient at steady state and this system yields much lower shear stress of 0.3(dynes/$\textrm{cm}^2$) than that of 5-8(dynes/$\textrm{cm}^2$) by conventional agitation systems.

• Bulletin of the Korean Chemical Society
• /
• v.28 no.9
• /
• pp.1515-1518
• /
• 2007

A preparative method of Me2SCuX (X = Cl, Br) via one-pot reaction was developed from common chemicals such as benzyl halide, DMSO, and Cu(0). In the reaction mixture small molecules such as methyl halide and dimethyl sulfide etc. were efficiently generated in situ, resulting in the formation of Me2SCuX. Oxidation of Cu(0) in reacting with both benzyl halide (starting material) and methyl halide (in situ generated product) to Cu(I) followed by complexing with dimethyl sulfide (in situ generated product) could explain the formation of Me2SCuX. In particular we found out that heterogeneous reaction of Me2SCuX and arenediazonium tetrafluoroborate in acetonitrile was so effective to afford corresponding bromoaromatics under mild conditions.