• Korean Journal of Animal Reproduction
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• v.14 no.2
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• pp.101-106
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• 1990

These mxperiments were carried out to investigate the effect of rabbit anti-bovine cumulus cell antibodies on in vitro fertilization and following development of bovine follicular oocytes matured in vitro. The bovine ovaries were obtained at a slaughter house and the follicular oocytes surrounded by cumulus cells were collected by puncturing follicles with 2~6mm of diameter. Bovine oocytes were matured in vitro for 24~26hrs in a CO2 incubator with 5% CO2 in air at 39$^{\circ}C$ and subsequently cultured in medium containing cumulus cell antibody for 1 hour. The medium used for maturation was TCM-199 supplemented with hormones, pyruvate, FCS and antibiotics. Epididymal spermatozoa were capacitated by in vitro culture for 2~3 hrs in BO solution 10~15 matured oocytes into the suspension of capacitated spermatozoa. Six hour after insemination the eggs were transferred to TCM-199 supplemented with FCS(10%) and then cultured for 7 days. The results obtained in these experiments were summarized as follows : 1. When the follicular oocytes matured in vitro were treated with antibody to intact cumulus cells, the fertilization rate of cumulus intact and removed oocytes was ranged to 45.0 to 53.7%. These value is slightly lower than that(64.3%) of follicular oocytes not treated with the antibody, and increased frequency of both male and female pronuclear formation was found in cumulus intact oocytes cultred in medium without the antibody(p<0.05). 2. The fertilization rate of cumulus intact and removed oocytes treated with antibody to solubilized cumulus cells was ranged 45.0 to 52.5%, significantly lowre than that(62.8%) of oocytes cultured in antibody free medium, and increased frequency of ova with male and female pronuclei was found when cumulus cells were present(p<0.05). 3. The rates of cumulus cell intact and removed oocytes developed to 8-, 16-cell and morula or blastocyst after treatment of intact and solubilized cumulus cell antibody were ranged 7.1 to 14.5, 2.9 to 5.9 and 1.5 to 2.9%, respectively, slightly lower than 18.6, 10.0 and 8.6% of cumulus intact oocytes cultured in medium without the antibody. The results of this stduy indicate that cumulus cells promote not only normal fertilization with proper pronuclear formation, but embryo development and that the beneficial effect of cumulus cell to the pronuclear formation and embryo development is blocked by the action of antibody to cumulus cell.

• Plant Biotechnology Reports
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• v.2 no.2
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• pp.163-169
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• 2008

Roots of Ophiorrhiza prostrata D. Don serve as a rich source of camptothecin (CPT), an anticancer drug. Because of the large-scale collection of its roots, the plant has become a threatened species. The present study accomplishes the induction of adventitious roots as a means for the production of CPT as well as for the large-scale propagation of this anticancer drug plant using leaf and internode explants. The biomass yield and CPT content of adventitious roots induced from different explants were compared to roots developed on ex vitro rooted stem cuttings. Adventitious roots were produced on half-strength Murashige and Skoog (MS) medium supplemented with $10.74{\mu}M$ ${\alpha}-naphthaleneacetic$ acid and $2.32{\mu}M$ kinetin at mean fresh weights of 0.753, 0.739 and 0.748 g roots from leaf, internode and shoot, respectively. CPT yield from in vitro derived roots after 50, 80 and 120 days of incubation (0.028, 0.06 and 0.1% dry weight, respectively) was not significantly different from those harvested at the same age from ex vitro rooted (0.03, 0.06 and 0.13%, respectively) stem cuttings. CPT from subcultured roots derived from solid (0.08%) medium was lower than from suspension culture medium (0.12%). Subsequent cultures of the adventitious roots showed a stable production of CPT (0.16%). The yield of CPT from 360-day-old plant-derived roots was 0.19%. Elicitation using methyl jasmonate and acetyl salicylic acid exhibited no enhancement in CPT yield. In vitro propagation through direct shoot regeneration was achieved from the adventitious roots upon transfer to MS medium with $8.87{\mu}M$ $N^6-benzyladenine$ (BA) and $2.46{\mu}M$ indole-3-butyric acid (IBA) with a mean of 21.2 shoots per culture in 50 days. The shoots upon subculture on medium having the same level of BA and IBA underwent rapid proliferation. The shoots transferred to field conditions after in vitro rooting exhibited 95% survival. Adventitious root induction, from leaf and internode explants, enables the feasible production of CPT as well as the large-scale rapid propagation of this species which can safeguard it from extinction.

• Korean Journal of Plant Tissue Culture
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• v.24 no.3
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• pp.183-188
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• 1997

Callus from scale segments of Lilium longiflorum 'Gelia' was effectively induced and maintained from unorganized tissue on the semi-solid medium by 0.42% Bacto agar with MS basal salts and vitamins of SH medium supplemented with 0.5 mg/L 2, 4-D, 1.0 mg/L NAA, 0.3 mg/L BA, and 3% sucrose. More than 5% of high sucrose level had inhibiting effect on regeneration capacity of formed callus and decreased callus growth. Various combinations of nitrogen did not effective to proliferate the ELC (Embryogenic-like callus), but friability of callus was increased in the medium containing only nitrate as nitrogen source. 5 mL conditioned medium into 30 mL fresh medium was good for cell growth. However friable cell aggregates during suspension culture had to form hard callus which hindered to establish suspention culture system. Addition of 2 g/L casein hydrolysate increased callus growth and friability of the hard callus. As a result of anatomical observation of callus, organogenesis such as shoots, roots and bulblets was independently induced from callus tissue. Somatic embryogenesis from callus tissue could be observed with low frequency.

• Journal of Plant Biotechnology
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• v.49 no.4
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• pp.356-361
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• 2022

Bacillus sp. is a useful strain for agriculture because it promotes plant growth and controls plant pathogens through a variety of mechanisms. In this study, we obtained a microbial preparation with a high number of viable cells by culturing newly isolated soil bacteria on an optimized medium. Subsequently, we applied this preparation to lettuce to enhance its growth and yield. First, B. amyloliquefaciens ISP-5 was isolated from soil. Next, optimization of culture medium was carried out using 5 L scale fermenters. When culturing B. amyloliquefaciens ISP-5 on this optimized medium, the number of viable cells was approximately 1000 times higher than that obtained from culturing on the commercial medium. Afterwards, the plant growth promotion properties of the ISP-5 strain were evaluated using lettuce as a test plant. Foliar spray treatment of lettuce was carried out by inoculating half the standard concentration suspension (0.5 × 10⁷ cfu/ml). As a result, leaf width increased by 8.6% and leaf length increased by 12.9% compared to the control group. Live weight also increased by 24.2% and dry weight by 23.9%. Considering the results from field test, B. amyloliquefaciens ISP-5 showed potential as a plant growth-promoting bacteria.

• Korean Journal of Animal Reproduction
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• v.25 no.4
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• pp.317-325
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• 2001

This study investigated the effect of ferrous sulfate (Fe$^{2+}$) and/or ascorbic acid (Asc) on fertilizing ability in vitro of frozen-thawed boar spermatozoa. Using chlortetracycline (CTC) fluorescence, the spermatozoa was treated in preincubation medium with control, Fe$^{2+}$(1 mM), Asc (0.5 mM) and Fe$^{2+}$Asc to assessed for acrosome reaction, and the oocyte penetration test to determine whether the Fe$^{2+}$ and/or Asc can promote the penetration ability in vitro. When frozen-thawed spermatozoa was washed with preincubation medium, there were significantly (P ＜ 0.05) more acrosome-reacted in medium with Fe$^{2+}$Asc (38%) than control (27%). The penetration rates were also significantly (P ＜ 0.05) higher in medium with Fe$^{2+}$Asc (76%) than control (55%). Next, the lipid peroxidation of sperm was evaluated on the basis of malondialdehyde production following same treatments. The addition of Fe$^{2+}$Asc to sperm suspension increases the formation of malondialdehyde. However, there were not significantly different under the all conditions. The sperm suspension were also treated with control, Fe$^{2+}$, Asc and Fe$^{2+}$/Asc and assayed for sulfhydry1(-SH) group content. In the Fe$^{2+}$/Asc group, sperm-SH group were higher than another groups. In spermatozoa treated with Fe$^{2+}$ and/or Asc, however, no changes in sperm -SH-groups were detected when compared to controls. In another experiment, the activity of sperm binding to zona pellucida was evaluated through binding to salt-stored porcine oocytes. In control and Asc treatment groups, sperm binding to zona pellucida were significantly (P ＜ 0.05) higher than in medium with Fe$^{2+}$. On the other hand, there is not a significant increase in binding to zona pellucida with spermatozoa treated by Fe$^{2+}$/Asc. In summary, the present study suggests that Fe$^{2+}$/Asc causes an enhancement in fertilizing ability that is associated with penetration rate increased without change of spermatozoa binding capacity to homologous zona pellucida.o homologous zona pellucida.

• Korean Journal of Environmental Agriculture
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• v.11 no.3
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• pp.215-223
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• 1992

A metabolic study has been conducted to investigate the conversion of pentachlorophenol(PCP) to water soluble metabolites in soybean and rice cell suspension cultures as well as in intact rice plants. PCP in plant cells was found to be exclusively transformed into water soluble metabolites. The relative rate of the metabolic conversion of PCP in decreasing order was soybean cultures > rice cultures > rice plants. Also observed was that, the older the cultures grown, the lower the conversion rate was. Primary water soluble metabolites isolated from both the 5 day old soybean and 8 day old rice cells were specifically hydrolyzed only by ${\beta}$-glucosidic linkage specific glucosidase, suggesting that the metabolites are ${\beta}$-glucose conjugates. The amount of glucose conjugates was increased with increasing time of incubation of PCP up to 24 hr in both soybean and rice cultures; Thereafter, it was decreased progressively. Most of the glucose conjugates were further metabolized to more polar conjugates in cells, but a portion of them was excreted into the culture medium.

• Korean Journal of Agricultural Science
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• v.20 no.1
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• pp.34-42
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• 1993

Effects of salt concentrations on the cell growth and the pigment accumulation were investigated in cell suspension culture of Vitis vinifera and Phytolacca americana L.. The growth pattern of vine cell in control was showed the normal exponential growth pattern, but in the dilution media delay the exponential growth pattern from 4 to 8 days after culture. Maximal accumulation of anthocyanin was observed at 12 days after culture in all treatments. In cell suspension culture of Phytolacca, accumulation of betacyanin occurred in parallel with the cell growth pattern and maximal accumulation of betacyanin was observed after 8 days of culture. In the vine cell culture, the cell growth was showed the peak at 87.6mM of sucrose in the medium and reduced at over this concentration. Maximal anthocyanin accumulation was showed at 146mM of sucrose. In the higher concentrations of sucrose, the cell growth was rapidly decreased, but the accumulation of anthocyanin was not. Otherwise, in case of Phytolacca cell culture, betacyanin accumulation was showed in parallel with the cell growth increased with sucrose concentration. It was suggested that the anthocyanin of vine and the betacyanin of Phytolacca were controlled by different mechanisms.

• Polymer(Korea)
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• v.30 no.6
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• pp.505-511
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• 2006

We tried to prepare polystyrene composite particles containing carbon black by suspension polymerization with water as a reaction medium. Hydrophobic silica was selected as a stabilizer and oil-soluble azobisisobutyronitrile (AIBN), as an initiator. All polymerization reactions were carried out at a fixed temperature of $75^{\circ}C$. Stabilizer concentration was varied from $0.17{\sim}3.33wt%$ compared to water, where particles with $7.96{\mu}m$ in average diameter were obtained at 1.57 wt% of stabilizer. Increase in divinylbenzene concentration, as a crosslinking agent, from $0.1{\sim}1.0 wt%$ compared to monomer exhibited a large increase in average particle diameter Incorporation of 1wt% of carbon black compared to monomer produced an increase in average diameter It is speculated that viscosity lower than that necessary to induce even dispersion of carbon black particles led to poor dispersion, and as a result, large particles. For a styrene mixture containing 3 wt% carton black compared to monomer, enhanced dispersion due to an increase in carbon black concentration reduced average particle diameters. For styrene mixtures containing 1 and 3 wt% carbon black compared to monomer, preparticles before polymerization and polymer composite particles after polymerization showed a similar tendency towards particle formation. When carbon black concentration compared to monomer was increased to 5 and 7 wt%, styrene mixtures exhibited a large increase in viscosity and thus better dispersion of carbon black particles, which led to a decrease in preparticle diameters. However, these particles experienced agglomeration in the polymerization process, and polystyrene composite particles increased in average diameter.

• 한국생물공학회:학술대회논문집
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• 2001.11a
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• pp.193-196
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• 2001

IL-18. formerly known as IGIF(interferon -gamma inducing factor), is structurally IL-l related but functionally IL-12 related pro-inflammatory cytokine. The human IL -18(hIL-lS), like IL-$1{\beta}$, is synthesized as a biologically inactive precursor of 24kDa lacking a signal peptide, and then cleaved into an active mature form by cystein protease IL-$1{\beta}$ converting enzyme (ICE: caspase- 1), We tested if the mature hIL -18 can be expressed and secreted into culture medium by transforming the forming gene construct consisting of a mature hIL-18 gene fused to signal peptide of rice amylase lA. Secondly, we were tested if the pro- IL-18 could be processed into a biologically active form by caspase-l like protease in plant. Cell suspension culture was established from the leaf-derived calli of transgenic tobacco plant. Southern and Northern blot analysis indicated the expression of both pro-hIL-18 and mature hIL-18 plant cells. Western blot analysis introduced the protein products of pro- hIL -18 and mhIL -18 were observed in transigenic cell lines. In addition, the molecular size of recombinant pro-hILl-18 and mhIL-18 were estimated to be 24kDa and 18kDa, respectively. ELISA revealed that the amount of pro- hIL -18 was 1.3ug per gram of fresh weight calli. Moreover, the presence of mhIL-18 was detected in the culture medium and it appeared to be 25ug/L.

• KSBB Journal
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• v.15 no.4
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• pp.366-369
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• 2000

The soluble paclitaxel was found in the supernatant of the plant cell cultures of Taxus chinensis, The percentage of soluble paclitaxel depends on paclitaxel concentration in bioreactor. As paclitaxel concentration decreases the percentage of soulbe paclitaxel increases. it is therefore important to develop a new process for the recovery of soluble paclitaxel. The use of hydrophobic resin HP20 gives nearly perfect recovery of paclitaxel in supernatant. The resin was more effective in treatment of th cell and debris free filtrate probably because of the reduced solids content In this case 3 g.l resin and 1 day reaction were enough for recovery the soluble paclitaxel in medium.