• KSBB Journal
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• v.22 no.5
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• pp.297-304
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• 2007

For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

• Journal of the Korea Concrete Institute
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• v.19 no.4
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• pp.515-525
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• 2007

In this investigation, results of laboratory tests on four reinforced concrete flat plate interior connections with elongated rectangular column support which has been used widely in tall residential buildings are presented. The purpose of this study is to evaluate an effect of column aspect ratio (${\beta}_c={c_1}/{c_2}$=side length ratio of column section in the direction of lateral loading $(c_1)$ to the direction of perpendicular to $c_1$) on the hysteretic behavior under earthquake type loading. The aspect ratio of column section was taken as $0.5{\sim}3\;(c_1/c_2=1/2,\;1/1,\;2/1,\;3/1)$ and the column perimeter was held constant at 1200mm in order to achieve nominal vertical shear strength $(V_c)$ uniformly. Other design parameters such as flexural reinforcement ratio $(\rho)$ of the slab and concrete strength$(f_{ck})$ was kept constant as ${\rho}=1.0%$ and $f_{ck}=40MPa$, respectively. Gravity shear load $(V_g)$ was applied by 30 percent of nominal vertical shear strength $(0.3V_o)$ of the specimen. Experimental observations on punching failure pattern, peak lateral-load and story drift ratio at punching failure, stiffness degradation and energy dissipation in the hysteresis loop, and steel and concrete strain distributions near the column support were examined and discussed in accordance with different column aspect ratio. Eccentric shear stress model of ACI 318-05 was evaluated with experimental results. A fraction of transferring moment by shear and flexure in the design code was analyzed based on the test results.

• Korean Journal of Poultry Science
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• v.28 no.2
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• pp.135-142
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• 2001

A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

• Journal of the Korea Concrete Institute
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• v.25 no.5
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• pp.485-496
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• 2013

In recent years, frequent terror or military attacks by explosion or impact accidents have occurred. Examplary case of these attacks were World Trade Center collapse and US Department of Defense Pentagon attack on Sept. 11 of 2001. These attacks of the civil infrastructure have induced numerous casualties and property damage, which raised public concerns and anxiety of potential terrorist attacks. However, a existing design procedure for civil infrastructures do not consider a protective design for extreme loading scenario. Also, the extreme loading researches of prestressed concrete (PSC) member, which widely used for nuclear containment vessel, gas tank, bridges, and tunnel, are insufficient due to experimental limitations of loading characteristics. To protect concrete structures against extreme loading such as explosion and impact with high strain rate, understanding of the effect, characteristic, and propagation mechanism of extreme loadings on structures is needed. Therefore, in this paper, to evaluate the impact resistance capacity and its protective performance of bi-directional unbonded prestressed concrete member, impact tests were carried out on $1400mm{\times}1000mm{\times}300mm$ for reinforced concrete (RC), prestressed concrete without rebar (PS), prestressed concrete with rebar (PSR, general PSC) specimens. According to test site conditions, impact tests were performed with 14 kN impactor with drop height of 10 m, 5 m, 4 m for preliminary tests and 3.5 m for main tests. Also, in this study, the procedure, layout, and measurement system of impact tests were established. The impact resistance capacity was measured using crack patterns, damage rates, measuring value such as displacement, acceleration, and residual structural strength. The results can be used as basic research references for related research areas, which include protective design and impact numerical simulation under impact loading.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.201-209
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• 2014

This study was carried out in order to develop a biological control of anthracnose of red pepper caused by fungal pathogens. In particular, this study focuses on the Colletotrichum species, which includes important fungal pathogens causing a great deal of damage to red pepper. Antagonistic bacteria were isolated from the soil of pepper fields, which were then tested for biocontrol activity against the Colletotrichum gloeosporioides anthracnose pathogen of pepper. Based on the 16S rRNA sequence analysis, the isolated bacterial strain CS-52 was identical to Bacillus sp. The culture broth of Bacillus sp. CS-52 had antifungal activity toward the hyphae and spores of C. gloeosporioides. Moreover, the substances with antifungal activity were optimized when Bacillus sp. CS-52 was grown aerobically in a medium composed of 0.5% glucose, 0.7% $K_2HPO_4$, 0.2% $KH_2PO_4$, 0.3% $NH_4NO_3$, 0.01% $MnSO_4{\cdot}7H_2O$, and 0.15% yeast extract at $30^{\circ}C$. The inhibition of spore formation resulting from cellulase, siderophores, and indole-3-acetic acid (IAA), were produced at 24 h, 48 h, and 72 h, respectively. Bacillus sp. CS-52 also exhibited its potent fungicidal activity against anthracnose in an in vivo test, at a level of 70% when compared to chemical fungicides. These results identified substances with antifungal activity produced by Bacillus sp. CS-52 for the biological control of major plant pathogens in red pepper. Further studies will investigate the synergistic effect promoting better growth and antifungal activity by the formulation of substances with antifungal activity.

• Horticultural Science & Technology
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• v.32 no.6
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• pp.872-878
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• 2014

This study aimed to enhance storage and freshness of strawberry fruits and foliage vegetables by spray treatment with Chlorella vulgaris as a bio-fertilizer. The tested strain, C. vulgaris CHK0008, was isolated from an organically cultivated rice paddy and identified as C. vulgaris by its morphology and 18S rDNA and 23S rDNA sequence homology. We successfully cultured C. vulgaris CHK0008 in BG11 modified medium (BG11MM) and adjusted $2.15{\times}10^6cell/mL$ C. vulgaris CHK0008 to one OD value by measuring the optical density at 680 nm using a UV-vis spectrophotometer. The soluble solid content of 'Seolhyang' and 'Yukbo' strawberry fruits treated by spray application with C. vulgaris CHK0008 was enhanced by 22.2% and 11.5% respectively, compared to untreated controls. Additionally, the decay rates of treated 'Seolhyang' and 'Yukbo' strawberry fruits decreased 63.8% and 74.4% respectively, compared to untreated control. Surface color changes and chlorosis of leaves in leaf vegetables such as lettuce, kale, red ornamental kale, white ornamental kale and beet were observed in samples treated with water spray for 10 days after cold storage. However, the decay rate of leafy vegetables treated with foliar application of 25% C. vulgaris CHK0008 liquid culture was significantly decreased compared to that of the untreated control during storage at $4^{\circ}C$.

• Tunnel and Underground Space
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• v.30 no.3
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• pp.256-269
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• 2020

The objective of this study is to evaluate the stress thresholds in crack development and the corrected fracture toughness of KURT granite under dry and saturated conditions. The stress thresholds were identified by calculation of inelastic volumetric strain from an uniaxial compression test. The corrected fracture toughness was estimated by using the Level II method (Chevron Bend specimen), suggested by ISRM (1988), in which non-linear behaviors of rock was taken into account. Average crack initiation stress(σ_ci) and crack damage stress(σ_cd) under a dry condition were 91.1 MPa and 128.7 MPa. While, average crack initiation stress(σ_ci) and crack damage stress(σ_cd) under a saturated condition were 58.2 MPa and 68.2 MPa. The crack initiation stress and crack damage stress of saturated ones decreased 36% and 47% respectively compared to those of dry specimens. A decrease in crack damage stress is relatively larger than that of crack initiation stress under a saturated condition. This indicates that the unstable crack growth can be more easily generated because of the saturation effect of water compared to the dry condition. The average corrected fracture toughness of KURT granite was 0.811 MPa·m^0.5. While, the fracture toughness of saturated KURT granite(K_CB) was 0.620 MPa·m^0.5. The corrected fracture toughness of rock in saturated condition decreases by 23.5% compared to that in dry condition. It is found that the resistance to crack propagation decreases under the saturated geological condition.

• The Korean Journal of Mycology
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• v.13 no.3
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• pp.157-168
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• 1985

To investigate nutritive values of a feed fermented with basidiomycetes, among the isolated strains, Lyophyllum decastes (Fr.) Sing. was found with the greatest enzyme productivity and rapid mycelial growth in rice straw medium. Optimum temperature, pH and moisture content for mycelial growth and enzyme production of the strain were $25{\sim}30^{\circ}C,\;pH\;4.0{\sim}7.0\;and\;70{\sim}75\;%$, respectively. Fifteen days of culture were required for the highest enzyme productivity. Among the sub-materials added, $30{\sim}40\;%$ of rice bran and $10{\sim}20\;%$ of defatted perilla seeds were effective for the enzyme production, but caused a reduced mycelial growth. The greatest effect of an addition of inorganic salts was obtained with $0.36{\sim}0.72\;%\;of\;(NH_4)_2HPO_4$. When 40 mesh or smaller rice straw and steam treatment at $0.5\;kg/cm^2$ were used, the mycelial growth decreased, whereas the enzyme production increased. The mycelial growth and enzyme production increased when $Ca(OH)_2$ was used as the alkali treatment, but decreased with increasing concentration of NaOH. As the fermentation proceeded, the amounts of ash, reducing sugar and total nitrogen increased, but cellulose, lignin and pentosan decreased. When the rice straw was treated with alkali, the amounts of ash, total nitrogen and lignin decreased, but reducing sugar and cellulose increased. At higher NaOH concentration, the variation become greater. The in vitro dry matter digestibility of the products increased from 55.03 % at the beginning of the fermentation to 62.72 % at 45 days after fermentation. The most effective alkali treatment on the digestibility of rice straw was KOH followed by NaOH. However, the digestibility increased with increasing concentration of NaOH. The digestibility of pretreated with alkali increased after fermentation as well.

• Journal of Biomedical Engineering Research
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• v.24 no.5
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• pp.401-410
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• 2003

This study investigates the biomechanical efficacies of various cement augmentation techniques with or without pressurization for varying degrees of osteoporotic femur. For this study, a biomechanical analysis using a finite element method (FEM) was undertaken to evaluate surgical procedures, Simulated models include the non-cemented(i.e., hip screw only, Type I), the cement-augmented(Type II), and the cemented augmented with pressurization(Type III) models. To simulate the fracture plane and other interfacial regions, 3-D contact elements were used with appropriate friction coefficients. Material properties of the cancellous bone were varied to accommodate varying degrees of osteoporosis(Singh indices, II∼V). For each model. the following items were analyzed to investigate the effect surgical procedures in relation to osteoporosis of varying degrees : (a) von Mises stress distribution within the femoral head in terms of volumetric percentages. (b) Peak von Mises stress(PVMS) within the femoral head and the surgical constructs. (c) Maximum von Mises strain(MVMS) within the femoral head, (d) micromotions at the fracture plane and at the interfacial region between surgical construct and surrounding bone. Type III showed the lowest PVMS and MVMS at the cancellous bone near the bone-construct interface regardless of bone densities. an indication of its least likelihood of construct loosening due to failure of the host bone. Particularly, its efficacy was more prominent when the bone density level was low. Micromotions at the interfacial surgical construct was lowest in Type III. followed by Type I and Type II. They were about 15-20% of other types. which suggested that pressurization was most effective in limiting the interfacial motion. Our results demonstrated the cement augmentation with hip screw could be more effective when used with pressurization technique for the treatment of intertrochanteric fractures. For patients with low bone density. its effectiveness can be more pronounced in limiting construct loosening and promoting bone union.

• Applied Microscopy
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• v.7 no.1
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• pp.21-43
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• 1977

This experiment was undertaken in order to localize the labeled dbcAMP (dibutyryl cyclic AMP) in oocytes whose development has been suppressed by cold dbcAMP for 6 or 19 hours in vitro. Mouse oocytes were obtained from the ovaries of 3-4 week old A strain female mice, by puncturing the Graafian follicles in the modified Krebs-Ringer bicarbonate salt solution under the dissecting microscope. Those oocytes which have intact germinal vesicle were cultured in the basic culture medium supplemented with 0.4% bovine serum albumin (BSA). Cultivation of the oocytes was carried out in a microtube developed by Cho (1974). The cultures were then incubated in a humidified 5% $CO_2$ incubator maintained at $37^{\circ}C$ for 6 or 19 hours (Donahue, 1968). DbcAMP was added to culture medium for a final concentration of 100ug/ml, and $^3H-dbc$ AMP (specific activity 13 Ci/mM) for a final concentration of $40{\mu}Ci/ml$ was also added to the medium. For electron microscopic autoradiography, those oocytes recovered from the culture were washed with phosphate buffer (pH 7.4), and immediately prefixed in a 2.5% glutaraldehyde overnight and postfixed for 2 hours at $4 ^{\circ}C$ in 1% osmium tetroxide in phosphate buffer with pH 7.4 (Palade, 1952). After fixation, the materials were dehydrated in graded alcohol series and embedded in Epon 812 mixture based on the standard procedures (Luft, 1961). The thin sections $600-700{\AA}$ thick were mounted on the grids of 200 meshes. The grids containing sections were coated with a nuclear emulsion Kodak NTB-3 and stored in a cold dark box (at $4^{\circ}C$) for 3 weeks. After exposure, the samples were developed with Kodak D-19 and stained with uranyl acetate and lead citrate. Routine observation was made with Hitachi HU-11E electron microsocope. The results of the observation were as followings: 1. It was found that the labeled dbcAMP penetrated the egg plasma membrane and dispersed at random in the cytoplasm. 2. It was also observed that most of the labeled dbcAMP was attached to microfibrillar lattices portion of the oocyte cytoplasm. There fore, it is presumed that the receptor of the dbcAMP is localized in the microfibrillar lattices of the oocyte. 3. It also seems that some other cell organells such as mitochondria, Golgi complex, cortical granules are not directly related to the action of the dbcAMP. 4. The labeled dbcAMP was neither observed in the membrane nor in the nucleus. Therefore, it seems that there is no relationship between the concentration of dbcAMP and the nuclear membranous permeability. 5. There was no difference in number of dbcAMP particles when oocytes were cultured for 6 hours and 19 hours. 6. However, it was observed that, in same of the oocytes suppressed in germinal vesicle by dbcAMP for 19 hours, cell organells were moved and concentrated to a small portion of the cytoplasm, and that the morphology of the organells greatly changed to an abnormal. form. Therefore, it is supposed that those oocytes were in the process of degeneration. From the above results, it is expected that dbcAMP penetrated the egg membrane and was bound to the receptor which seems to be located in the microfibrillar lattiees portion, and that this dbcAMP-receptor complex inhibited some enzyme system of the oocytes which are essential for the germinal vesicle breakdown.