The gene encoding $\beta$-agarase (agaB) which hydrolyzes $\beta$-1,4 linkages of agarose from Zobellia galactanivorans was cloned and fused to Saccharomyces cerevisiae mating factor alpha-1 secretion signal ($MF{\alpha}1$), in which the transcription of $MF{\alpha}1$-AgaB was under the control of AOX1 (alcohol oxidase 1, methanol inducible) promoter. The constructed plasmid pPIC-AgaB (9 kb) was integrated into HIS4 gene locus of Pichia pastoris genome. Successful integration was confirmed by performing colony PCR. The transformed cells showed red halos around its colonies in methanol agar plate by adding iodine solution, indicating the active expression of agaB in P.pastoris. By SDS-PAGE and zymographic analysis, the molecular weight of $\beta$-agarase was estimated to be a 53 kDa and about 15% N-linked glycosylation was occurred. The activity of extracellular $\beta$-agarase reached 1.34, 1.42 and 1.53 units/mL by inducing 0.1, 0.5, and 1% methanol, respectively, at baffled flask culture of P.pastoris GS115/pPIC-AgaB for 48 hr. Most of the enzyme activity was found in the extacellular fraction and the secretion efficiency showed 98%. Thermostability of recombinant $\beta$-agarase was also increased by glycosylation.
Kim, Young-Ro;Kim, Yong-Hyu;Hur, Myung-Joon;Woo, Jong-Myung
The Journal of Korean Institute of Electromagnetic Engineering and Science
/
v.24
no.3
/
pp.219-232
/
2013
This paper presents a microstrip antenna which recovers its resonant frequency and impedance shifted automatically by the approach of other objects such as hands. This can be used for telemetry sensor applications in the ultrahigh frequency(UHF) industrial, scientific, and medical(ISM) band. It is the key element that an frequency-reconfigurable antenna could be electrically controlled. This antenna is miniaturized by loading the folded plates at both radiating edges, and varactor diodes are installed between the radiating edges and the ground plane to control the resonant frequency by adjusting the DC bias asymmetrically. Using this voltage-controlled antenna and the micro controller peripheral circuits of reading the returned level, the antenna is designed and fabricated which recovers its resonant frequency and impedance automatically. Designed frequency auto recovering antenna is conformed to be recovered within a few seconds when the resonant frequency and impedance are shifted by the approach of other objects such as hand, metal plate, dielectric and so on.
Kim, Min Jeong;Shim, Chang Ki;Kim, Yong Ki;Hong, Sung Jun;Park, Jong Ho;Han, Eun Jung;Kim, Jin Ho;Kim, Suk Chul
The Plant Pathology Journal
/
v.31
no.3
/
pp.259-268
/
2015
This study investigated the chemical characteristics and microbial population during incubation of four kinds of aerated compost teas based on oriental medicinal herbs compost, vermicompost, rice straw compost, and mixtures of three composts (MOVR). It aimed to determine the effects of the aerated compost tea (ACT) based on MOVR on the growth promotion of red leaf lettuce, soybean and sweet corn. Findings showed that the pH level and EC of the compost tea slightly increased based on the incubation time except for rice straw compost tea. All compost teas except for oriental medicinal herbs and rice straw compost tea contained more ${NO^-}_3-N$ than ${NH^+}_4-N$. Plate counts of bacteria and fungi were significantly higher than the initial compost in ACT. Microbial communities of all ACT were predominantly bacteria. The dominant bacterial genera were analyzed as Bacillus (63.0%), Ochrobactrum (13.0%), Spingomonas (6.0%) and uncultured bacterium (4.0%) by 16S rDNA analysis. The effect of four concentrations, 0.1%, 0.2%, 0.4% and 0.8% MOVR on the growth of red leaf lettuce, soybean and sweet corn was also studied in the greenhouse. The red leaf lettuce with 0.4% MOVR had the most effective concentration on growth parameters in foliage part. However, 0.8% MOVR significantly promoted the growth of root and shoot of both soybean and sweet corn. The soybean treated with higher MOVR concentration was more effective in increasing the root nodule formation by 7.25 times than in the lower MOVR concentrations Results indicated that ACT could be used as liquid nutrient fertilizer with active microorganisms for culture of variable crops under organic farming condition.
Seo, Hye-Won;Park, Min-Suk;Lee, Swoo-Heon;Shin, Kyung-Jae
Journal of Korean Society of Steel Construction
/
v.22
no.5
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pp.477-485
/
2010
Various monitoring sensors are used to predict and detect structural damage. Smart sensors, such as glass-fiber sensors, PZT, and MEMS, among others, have replaced traditional sensors. They are now being used in many areas. This study aims to predict the damage by measuring the PZT voltage attached on the specimen by the applied impact load. In the experiment to detect damages in beam connection, simple $H-400{\times}200{\times}8{\times}13$ beams were spliced with bolts. The results of FFT between PZT sensor and accelrometer were compared to measure the sensitivity of the PZT sensor. The damage to the beam was presumed by loosening the bolt, and then the damage measurement was accompanied. Secondly, a steel $PL600{\times}65{\times}5.8$ plate beam was fabricated for the purpose of experimenting on damage measurement. Impact loading test on three different locations was carried out. Damage width varied between 6~42mm on both sides by cutting, using a steel saw. The ratio of frequencies before and after the damage was computed to quantify the damage level by using FFT, and the change in mode pattern with the increased damage was investigated to measure the damage.
In this study, the so-called repeated plate load bearing test (RPBT) used to get $E_{v2}$ values in order to check the degree of compaction of subgrade, and to get design parameters for determining the thickness of the trackbed foundation, is investigated. The test procedure of the RPBT method is scrutinized in detail. $E_{v2}$ values obtained from the field were verified in order to check the reliability of the test data. The $E_{v2}$ values obtained from high-speed rail construction sites were compared to converted modulus values obtained from resonant column (RC) test results. For these tests, medium-size samples composed of the same soils from the field were used after analyzing stress and strain levels existing in the soil below the repeated loading plates. Finite element analyses, using the PLAXIS and ABAQUS programs, were performed in order to investigate the impact of the strain influence coefficient. This was done by getting newly computed $I_z$ to get the precise strain level predicted on the subgrade surface in the full track structure; under wheel loading. It was verified that it is necessary to use precise loading steps to construct nonlinear load-settlement curves from RPBT in order to get correct $E_{v2}$ values at the proper strain levels.
Kim, Min-Gyun;Park, Hyun-Jae;Lee, Dong-Guen;Hwang, Hyun-Sik;Kim, Hyun-Su
Journal of the Earthquake Engineering Society of Korea
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v.13
no.5
/
pp.11-21
/
2009
Considering that the weight of a biaxial hollow slab system is not increased with an incremental increase in its thickness, and that the flexural stiffness of a biaxial hollow slab is not significantly lower than that of a general solid slab, there has been a growing need for biaxial hollow slab systems, because long span structures are in great demand. In a long span structure, the problem of vibration of floor slabs frequently occurs, and the dynamic characteristics of a biaxial hollow slab system are quite different from the conventional floor systems. Therefore, in this study, the floor vibration of a biaxial hollow slab system subjected to walking load is investigated in comparison with a conventional floor slab system. For the efficiency of time history analysis, an equivalent plate slab model that can precisely represent the dynamic behavior of a biaxial hollow slab system is used. From the analytical results, it was determined that vibration of a biaxial hollow slab system subjected to walking load is evaluated as "office-level vibration," according to the classifications of the architectural institute of Japan and ANSI.
Orthodontic tooth movement in response to orthodontic force results from actions of osteoclasts and osteeoblasts in the cell level. Convincing evidence has now been provided to support the view that osteoclasts are derived from mononuclear cells that originate in the bone marrow or other hematopoietic organs and they migrate to the bones via vascular routes. Nitric oxide(NO), which accounts for the biological properties of endothelium-derived relaxing factor(EDRF), is the endogenous stimulator of soluble guanylate cylase. The discovery of the formation of nitric oxide(NO) from L-arginine in mammalian tissues and its biological roles has, in the last 7 years, thrown new light onto many areas of research. Data from experiments in vitro showed that N-metyl-L-arginine(L-NMA) and L-nitro-L- arginine(L-NAME) are competitive inhibitors of nitric oxide synthase. This study suggest that the multinucleated cells in our culture have characteristics of osteoclasts and that the potential bone cell activity of nitric oxide in vitro may be mediated in part by stimulation of marrow mononuclear cells to form osteoclast-like cells. Bone marrow cells were obtaineed from tibia of 19-days old chick embryo. After sacrifice, tibia was quickly dissected and the bone were then split to expose the medullary bone. The cells were attached for 4 hours and the nonadherent cells were collected. Marrow cells weere cultured in 96-well plate in medium 199. To examine the number of TRAP-positive multinucleated cells(MNCs), $10^{-8}\;M\;Vit=D_3$ and various concentration of L-NMA and L-NAME weere added at the beginning of cultures and with each medium change. After 7 days of culture. tartrate-resistant acid phosphatase(TRAP) staining was performed for microscopic evaluation. Cells haying more than three nuclei per cell were counted as MNCs. The obsrved results were as follows;1. 1,25-dihydroxyvitamine $D_3$ stimulated the osteoclast-like multinucleated cells in cultures of chick embryo bone marrow. 2. Nitric oxide synthase inhibitors(NOSI ; N-NMA, N-NAME) stimulated the osteoclast-like cells in cultures of chick embry bone marrow. 3. 1,25-dihydroxyvitamine$D_3$ and nitric oxide synthase inhibitors did not appear to have additive effect on the generation of TRAP-positive MNCs. These results suggest that nitric oxide synthase inhibitors may stimulate the osteoclast-like multinucleated cell formation and fusion in cultures of chick bone marrow.
The purpose of this study was to evaluate hazard factors in the steps of production, holding and assembly and service of jabchae that were served by contracted management in school meal foodservice. And method of control with the Hazard Analysis Critical Control Point (HACCP)program was suggested. The hazard factors in jabchae product had come from the temperature, time, growth of microbiological equipment and utensils. Almost of raw materials, prepreparation and storage, cooking, holding, serving established the critical control point. It has shown that the microbiological quality of raw materials for jabchae was a little inferior at the time of receiving based on the total plate count(10$^3$∼10 $^{5}$ ), coliform(0∼10$^3$). However microbiological quality become acceptable level by washing and a few disinfection method. Microbiological growth has increased in 3hr at room temperature up to 5.1${\times}$10$^2$ after preparation of jobchae. In conclusion, it is extremly important to reduce hoiding and serving time and to avoid treating food within the danger zone for food safeness. In addition, the prevention of cross-contamination during mixing the ingredients with improper equipment and with insanitary treatments by workers was also important to keep the food safe in this foodservice facility.
Kim, Mi-Ae;Yoo, So-Young;Kim, Hwa-Sook;Kook, Joong-Ki;Lim, Sung-Hoon;Yoon, Young-Jooh;Kim, Kwang-Won
The korean journal of orthodontics
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v.35
no.1
s.108
/
pp.51-59
/
2005
The aim of this study was to compare the species and biotypes of mutans streptococci isolated from dental plaques sampled from the interfaces between the bracket aid tooth surface and smooth tooth surfaces In orthodontic patients. Dental plaque was collected from the interfaces between brackets aid teeth (test group), and from smooth tooth surfaces distant from brackets by more that 2mm (control group). The dental plaque collected by a sterilized curette was transferred into a vial of 1 X PBS. The sample in the vial was vigorously vortexed for 1 min and plated ou mitis-salivarius bacitracin (MSB) agar plate using cotton tips. The agar plates were incubated at $37^[\circ}C$ in a candle jar for 2 days, and again incubated for 1 more day at anambient temperature Individual colonies were cultured in TH broth at $37^[\circ}C\;CO_2$ incubator. The PCR-RFLP based on dextranase gene was performed for the identification of mutans streptococci at the species-level For biotyping of mutans streptococci, biochemical tests were performed There was no significant difference of the species of mutans streptococci isolated from both test and control groups However, the biotypes of the mutans streptococci isolated from test and control groups were different. These results may offer the basic data to verify the relationship between the mutans streptococci biotype and enamel decalcification or dental caries in orthodontic patients with fixed appliances.
Journal of the korean academy of Pediatric Dentistry
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v.24
no.1
/
pp.204-219
/
1997
The use of fluoride is one of the most effective methods for caries prevention. Fluoridation of public water supply has been recognized, for many years, as an effective way to reduce dental caries. The fluoride supplement has been recommended when the natural fluoride was unavailable or below the optimal range. However the mechanism of caries prevention by fluoride has not yet been clarified and it is well known that an overdose of fluoride results inacute and chronic toxicity, especially dental fluorosis. Fluoride mouthrinsing solution is widely used in dentistry due to its effectiveness in carrying anticariogenic action. Understanding the effects of fluoride mouthrinsing solution on human gingival fibroblasts will provide the safety rationale for its use during the caries preventive therapy. The purpose of this study was to evaluate the cytotoxic effect of fluoride mouthrinsing solution on the human gingival fibroblast in vitro. The human gingival fibroblasts were cultured from healthy gingiva on the extracted deciduous teeth of children. Cells were inoculated into a 24-well plate with $1{\times}10^4cells/well$ of medium at $37^{\circ}C$, 100% humidity, 5% $CO_2$ incubator for 24 hours. And the cells were counted by using the hemocytometer at each designed study. Human gingival fibroblasts were cultured in growth medium after one minute application range of 0.02%-0.2% NaF solution and 0.1% $SnF_2$ solution. The cells used in this study were between fifth to eighth passage number. The cell morphology was examined by inverted microscope and cell proliferation was measured by incorporating $[^3H]$-thymidine into DNA. DNA synthesis by human gingival fibroblasts was assessed by $[^3H]$-thymidine uptake assays while the cell activity was measured by MTT assay. Each concentrated fluoride mouthrinsing solution was estimated for its biocompatability with fibroblasts by the tissue culture technique. The results of this study were as follows : 1. It was observed that at 0.05%, 0.2% NaF mouthrinsing solution the cytoplasmic processes became globular. When 0.1% $SnF_2$ mouthrinsing solution was applied, the cytoplasmic process and cell morphology were disappeared. 2. DNA synthetic activity was reduced regardless of the concentration of the fluoride mouthrinsing solution. However, the result is statistically insignificant except 0.1% $SnF_2$ mouthrinsing solution(p<0.05). 3. Our results indicate that 0.02%, 0.05% concentrations of NaF mouthrinsing solution caused minimal cytotoxicity. But 0.2% NaF and 0.1% $SnF_2$ concentration were a significant difference between the cell activity in the experimental group and control group (p<0.05). 4. After appling 0.05% & 0.02% NaF fluoride mouthrinsing solution, cell activity was restored to the control groups level according to incubating time. The results suggest that direct exposure to fluoride solution inhibits gingival fibroblast activity. Therefore, for the most effective use of fluoride use, lowering the concentration of fluoride mouthrinsing is advisable because it maintains biocompatability and free ion in the oral fluid.
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