• Korean Journal of Microbiology
• /
• v.46 no.1
• /
• pp.21-26
• /
• 2010

As the causative agent of Anthrax, Bacillus anthracis causes an acute fatal disease in herbivores such as cattle, sheep, and horses as well as humans. The therapeutics and prevention of anthrax currently available are based on antibiotics and the live attenuated vaccine strains, which may be problematic due to the emergency of antibiotic resistant strains or residual virulence in those vaccine strains. Therefore, it has been required to develop novel therapeutics and vaccines which are safer and applicable to humans. Recently, the development of the multivalent vaccine targeting both spores and vegetative cells of B. anthracis along with anthrax toxin has been reported. In our attempts to screen potential candidates for those multivalent vaccines, the whole genomic expression library of B. anthracis was constructed in this study. To the end, the partial digests of the genomic DNA from B. anthracis (ATCC 14578) with Sau3AI were ligated with the inducible pET30abc expression vectors, resulting in approximately $1{\times}10^5$ clones in E. coli BL21(DE3). The redundancy test by DNA nucleotide sequencing was performed for the randomly selected 111 clones and found 56 (50.5%) B. anthracis genes, 17 (15.3%) vector sequences, and 38 (34.2%) unknown genes with no sequence homology by BLAST. An inducible expression of the recombinant proteins was confirmed by Western blot. Interestingly, some clones could react with the antiserum against B. anthracis. These results imply that the whole genomic library constructed in this study can be applied for analyzing the immunomes of B. anthracis.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.30 no.6
• /
• pp.465-473
• /
• 2004

Purpose : The essential triad for nerve regeneration is nerve conduit, supporting cell and neurotrophic factor. In order to improve the peripheral nerve regeneration, we used polyglycolic acid(PGA) tube and brain-derived neurotrophic factor(BDNF) gene transfected Schwann cells in sciatic nerve defects of SD rat. Materials and methods : Nerve conduits were made with PGA sheet and outer surface was coated with poly(lactic-co-glycolic acid) for mechanical strength and control the resorption rate. The diameter of conduit was 1.8mm and the length was 17mm Schwann cells were harvested from dorsal root ganglion(DRG) of SD rat aged 1 day. Schwann cells were cultured on the PGA sheet to test the biocompatibility adhesion of Schwann cell. Human BDNF gene was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into E1 deleted region of adenovirus shuttle vector, pAACCMVpARS. BDNF-adenovirus was multiplied in 293 cells and purified. The BDNF-Adenovirus was then infected to the cultured Schwann cells. Left sciatic nerve of SD rat (250g weighing) was exposed and 14mm defects were made. After bridging the defect with PGA conduit, culture medium(MEM), Schwann cells or BDNF-Adenovirus infected Schwann cells were injected into the lumen of conduit, respectively. 12 weeks after operation, gait analysis for sciatic function index, electrophysiology and histomorphometry was performed. Results : Cultured Schwann cells were well adhered to PGA sheet. Sciatic index of BDNF transfected group was $-53.66{\pm}13.43$ which was the best among three groups. The threshold of compound action potential was between 800 to $1000{\mu}A$ in experimental groups which is about 10 times higher than normal sciatic nerve. Conduction velocity and peak voltage of action potential of BDNF group was the highest among experimental groups. The myelin thickness and axonal density of BDNF group was significantly greater than the other groups. Conclusion : BDNF gene transfected Schwann cells could regenerate the sciatic nerve gap(14mm) of rat successfully.

• Korean Journal of Remote Sensing
• /
• v.27 no.2
• /
• pp.191-201
• /
• 2011

Soybean has widely grown for its edible bean which has numerous uses. Microwave remote sensing has a great potential over the conventional remote sensing with the visible and infrared spectra due to its all-weather day-and-night imaging capabilities. In this investigation, a ground-based polarimetric scatterometer operating at multiple frequencies was used to continuously monitor the crop conditions of a soybean field. Polarimetric backscatter data at L, C, and X-bands were acquired every 10 minutes on the microwave observations at various soybean stages. The polarimetric scatterometer consists of a vector network analyzer, a microwave switch, radio frequency cables, power unit and a personal computer. The polarimetric scatterometer components were installed inside an air-conditioned shelter to maintain constant temperature and humidity during the data acquisition period. The backscattering coefficients were calculated from the measured data at incidence angle $40^{\circ}$ and full polarization (HH, VV, HV, VH) by applying the radar equation. The soybean growth data such as leaf area index (LAI), plant height, fresh and dry weight, vegetation water content and pod weight were measured periodically throughout the growth season. We measured the temporal variations of backscattering coefficients of the soybean crop at L, C, and X-bands during a soybean growth period. In the three bands, VV-polarized backscattering coefficients were higher than HH-polarized backscattering coefficients until mid-June, and thereafter HH-polarized backscattering coefficients were higher than VV-, HV-polarized back scattering coefficients. However, the cross-over stage (HH > VV) was different for each frequency: DOY 200 for L-band and DOY 210 for both C and X-bands. The temporal trend of the backscattering coefficients for all bands agreed with the soybean growth data such as LAI, dry weight and plant height; i.e., increased until about DOY 271 and decreased afterward. We plotted the relationship between the backscattering coefficients with three bands and soybean growth parameters. The growth parameters were highly correlated with HH-polarization at L-band (over r=0.92).

• Journal of the Korean Society of Hazard Mitigation
• /
• v.3 no.3 s.10
• /
• pp.151-163
• /
• 2003

In this study, the algorithm of groundwater flow process was established for koreanized groundwater program development dealing with the geographic and geologic conditions of the aquifer have dynamic behaviour in groundwater flow system. All the input data settings of the 3-DFM model which is developed in this study are organized in Korean, and the model contains help function for each input data. Thus, it is designed to get detailed information about each input parameter when the mouse pointer is placed on the corresponding input parameter. This model also is designed to easily specify the geologic boundary condition for each stratum or initial head data in the work sheet. In addition, this model is designed to display boxes for input parameter writing for each analysis condition so that the setting for each parameter is not so complicated as existing MODFLOW is when steady and unsteady flow analysis are performed as well as the analysis for the characteristics of each stratum. Descriptions for input data are displayed on the right side of the window while the analysis results are displayed on the left side as well as the TXT file for this results is available to see. The model developed in this study is a numerical model using finite differential method, and the applicability of the model was examined by comparing and analyzing observed and simulated groundwater heads computed by the application of real recharge amount and the estimation of parameters. The 3-DFM model is applied in this study to Sehwa-ri, and Songdang-ri area, Jeju, Korea for analysis of groundwater flow system according to pumping, and obtained the results that the observed and computed groundwater head were almost in accordance with each other showing the range of 0.03 - 0.07 error percent. It is analyzed that the groundwater flow distributed evenly from Nopen-orum and Munseogi-orum to Wolang-bong, Yongnuni-orum, and Songja-bong through the computation of equipotentials and velocity vector using the analysis result of simulation which was performed before the pumping started in the study area. These analysis results show the accordance with MODFLOW's.

• Journal of Intelligence and Information Systems
• /
• v.17 no.4
• /
• pp.95-108
• /
• 2011

TThis is a study of the personalization method that intelligently adapts the level of clustering considering purchasing index of a customer. In the e-biz era, many companies gather customers' demographic and transactional information such as age, gender, purchasing date and product category. They use this information to predict customer's preferences or purchasing patterns so that they can provide more customized services to their customers. The previous Customer-Segmentation method provides customized services for each customer group. This method clusters a whole customer set into different groups based on their similarity and builds predictive models for the resulting groups. Thus, it can manage the number of predictive models and also provide more data for the customers who do not have enough data to build a good predictive model by using the data of other similar customers. However, this method often fails to provide highly personalized services to each customer, which is especially important to VIP customers. Furthermore, it clusters the customers who already have a considerable amount of data as well as the customers who only have small amount of data, which causes to increase computational cost unnecessarily without significant performance improvement. The other conventional method called 1-to-1 method provides more customized services than the Customer-Segmentation method for each individual customer since the predictive model are built using only the data for the individual customer. This method not only provides highly personalized services but also builds a relatively simple and less costly model that satisfies with each customer. However, the 1-to-1 method has a limitation that it does not produce a good predictive model when a customer has only a few numbers of data. In other words, if a customer has insufficient number of transactional data then the performance rate of this method deteriorate. In order to overcome the limitations of these two conventional methods, we suggested the new method called Intelligent Customer Segmentation method that provides adaptive personalized services according to the customer's purchasing index. The suggested method clusters customers according to their purchasing index, so that the prediction for the less purchasing customers are based on the data in more intensively clustered groups, and for the VIP customers, who already have a considerable amount of data, clustered to a much lesser extent or not clustered at all. The main idea of this method is that applying clustering technique when the number of transactional data of the target customer is less than the predefined criterion data size. In order to find this criterion number, we suggest the algorithm called sliding window correlation analysis in this study. The algorithm purposes to find the transactional data size that the performance of the 1-to-1 method is radically decreased due to the data sparity. After finding this criterion data size, we apply the conventional 1-to-1 method for the customers who have more data than the criterion and apply clustering technique who have less than this amount until they can use at least the predefined criterion amount of data for model building processes. We apply the two conventional methods and the newly suggested method to Neilsen's beverage purchasing data to predict the purchasing amounts of the customers and the purchasing categories. We use two data mining techniques (Support Vector Machine and Linear Regression) and two types of performance measures (MAE and RMSE) in order to predict two dependent variables as aforementioned. The results show that the suggested Intelligent Customer Segmentation method can outperform the conventional 1-to-1 method in many cases and produces the same level of performances compare with the Customer-Segmentation method spending much less computational cost.

• Proceedings of the Korean Society of Plant Pathology Conference
• /
• 1995.06b
• /
• pp.77-96
• /
• 1995

To obtain basic informations for the improvement of seed potato production in Korea, some etiological properties of potato virus Y(PVY) distributed in the major seed potato production area(Daekwanryeong) were characterized, and the nucleotide and amino acid sequences of the coat protein gene of the PVY strains isolated were analyzed. PVY strains in Daekwonryeong, an alpine area, were identified to be two strains, PVYo and PVYN by symptoms of indicator plants, and their distribution in potato fields was similar. Major symptom on potato varieties by PVY was grouped as either mosaic alone or mosaic accompanied with veinal necrosis in the lower leaves. The symptom occurrence of the two symptoms was similar with Irish Cobbler, but Superior showed a higher rate of mosaic symptom than the other. The PVY strain which was isolated from potato cv. Superior showing typical mosaic symptoms produced symptoms of PVY-O on the indicator plants of Chenopodium amaranticolor, Nicotiana tabacum cv. Xanthi nc and Physalis floridana, but no symptom o Capsicum annum cv. Ace. Moreover, results from the enzyme-linked immunosorbent assay with monoclonal and polyclonal antibodies showed that the isolated PVY reacts strongly with PYV-O antibodies but does not react specifically with PVY-T antibodies. The purified virus particles were flexious with a size of 730$\times$11nm. On the basis of the above characteristics, the strain was identified to be a PVY-O and named as of PVY-K strain. The flight of vector aphids was observed in late May, however, the first occurrence of infected plants was in mid June with the bait plants surrounded with PVY-infected potato plants and early July with the bait plants surrounded with PVY-free potato plants. PVY infection rates by counting symptoms on bait plants (White Burley) were 1.1% with the field surrounded with PVY-free potato plants and 13.7% the fields surrounded with PVY-infected potato plants, showing the effect of infection pressure. The propagated PVY-K strain on tobacco(N. sylvestris) was purified, and the RNA of the virus was extracted by the method of phenol extraction. The size of PVY-K RNA was measured to be 9, 500 nucleotides on agarose gel electrophoresis. The double-stranded cDNAs of PVY-K coat protein(CP) gene derived by the method of polymerase chain reaction were transformed into the competent cells of E. coli JM 109, and 2 clones(pYK6 and pYK17) among 11 clones were confirmed to contain the full-length cDNA. Purified plasmids from pYK17 were cut with Sph I and Xba I were deleted with exonuclease III and were used for sequencing analysis. The PVY-K CP gene was comprised of 801 nucleotides when counted from the clevage site of CAG(Gln)-GCA(Ala) to the stop codon of TGA and encoded 267 amino acids. The molecular weight of the encoded polypeptides was calculated to be 34, 630 daltons. The base composition of the CP gene was 33.3% of adenine, 25.2% of guanine, 20.1% of cytosine and 21.4% of uracil. The polypeptide encoded by PVY-K CP gene was comprised of 22 alanines, 20 threonines, 19 glutamic acids and 18 glycines in order. The homology of nucleotide sequence of PVY-K CP gene with those of PVY-O(Japan), PVY-T(Japan), PVY-TH(Japan), PVYN(the Netherlands), and PVYN(France) was represented as 97.3%, 88.9%, 89.3%, 89.6% and 98.5%, respectively. The amino acid sequence homology of the polypeptide encoded by PVY-K CP gene with those encoded by viruses was represented as 97.4%, 92.5%, 92.9%, 92.9%, and 98.5%, respectively.

• Nuclear Medicine and Molecular Imaging
• /
• v.43 no.5
• /
• pp.487-494
• /
• 2009

Purpose: Small size of recombinant scFv antibody has many advantages such as rapid blood clearances and improved targeting antibodies to tumor region. On the other hand owing to small size, number of amino group is insufficient in conjugation with chelator and fluorescence labeling. This study is to introduce poly lysine tag to the C-terminal end of scFv lym-1 sequence for fluorescence chelator conjugation. Materials and Methods: Poly lysine scFv lym-1 gene, cloned into pET-22b (+) vector, was expressed in E. coli BL21 (DE3) strain. Antibody purification was performed with Ni-NTA column and then size exclusion column chromatography. Expression and purification levels of poly lysine tagged scFv lym-1 antibody were confirmed by western blot analysis. I-124, I-125, I-131 and Tc-99m were used for radiolabeling of purified poly lysine scFv lym-1. Flow cytometry analysis of FIT( conjugated poly lysine scFv lym-1 was performed for confirmation of immunoreactivity of human Burkitt's lymphoma cells. Results: Poly lysine scFv lym-1 antibody was purified through two steps and identified as molecular weight of 48 KDa. Radiolabeling yields of I-124, I-125, I-131 and Tc-99m into poly lysine scFv lym-1 were >99%, >99%, >95% and >99%, respectively. Flow cytometry analysis of poly lysine scFv and scFv lym-1 was showed similar immunoreactivity to human Burkitt's lymphoma cells. Conclusion: Poly lysine tag was useful for the sufficient number of amino groups to scFv lym-1 antibody for chelator conjugation with minimizing loss of immunoreactivity.

• Journal of Biosystems Engineering
• /
• v.4 no.1
• /
• pp.1-23
• /
• 1979

The use of barley seeder attached to rotary tiller in the rural area has a significant meaning not only for the solution of labor peak season, but also for the increase of land utilization efficiency. The facts that presently being used barley seeders are all based on the mechanical principles of the reverse rotation, center drive and are all using forward rotating tine, which is used to be easily and heavily worn out when it rotates reversely, raise problem of recommending them to rural area in Korea. Therefore, the main objective of the study was to develop new type of rotary tine attachable to barley seeders. To attain the objective the following approaches were applied. (1) The kinematic analysis of reverse rotating barley seeders. (2) The studies on the soil bin and artificial soil. (3) The comparative experiment on the power requirement of prototype tine. The results obtained from the studies are summarized as follow: 1. The kinematic analysis of barley seeder attached to rotary tiller: The following results were obtained from the kinematic analysis for deriving general formulae of the motion and velocity characterizing the rotary tine of barley seeders presently being used by farmers. a) The position vector (P) of edge point (P) in the rotary tine of reverse rotating, center drive was obtained by the following formula. $$P=(vt+Rcos wt)i+Rsin wt j+ \{ Rcos \theta r sin \alpha cos (wt- \beta +\theta r) +Rsin \theta r sin \alpha sin (wt-\beta + \theta r) \} lk $$ b) The velocity of edge point $(P^')$ of reverse rotating, center drive rotary tine was obtained by the following formula. $$(P^')=(V-wR sin wt)i+(w\cdot Rcoswt)j + \{ -w\cdot Rcos \theta r\cdot sin \alpha \cdot sin (wt-\beta +\theta r) + w\cdot Rsin \theta r\cdot sin \alpha \cdot cos (wt- \beta + \theta r \} k $$ c) In order to reduce the power requirement of rotary tine, the angle between holder and edge point was desired to be reduced. d) In order to reduce the power requirement, the edge point of rotary tine should be moved from the angle at the begining of cutting to center line of machine, and the additional cutting width should be also reduced. 2. The studies on the soil bin and artificial soil: In order to measure the power requirement of various cutting tines under the same physical condition of soil, the indoor experiments Viere conducted by filling soil bin with artificially made soil similar to the common paddy soil and the results were as follows: a) When the rolling frequencies$(x)$ of the artificial soil were increased, the densIty$(Y)$ was also increased as follows: $$y=1.073200 +0.070780x - 0.002263x^2 (g/cm^3)$$ b) The absolute hardness $(Y)$ of soil had following relationship with the rolling frequencies$(x)$ and were increased as the rolling frequencies were increased. $$Y=37.74 - \frac {0.64 + 0.17x-0. 0054x^2} {(3.36-0.17x + 0.0054x^2)^3} (kg/cm^3)$$ c) The density of soil had significant effect on the cohesion and angle of internal friction of soil. For instance, the soil with density of 1.6 to 1.75 had equivalent density of sandy loam soil with 29.5% of natural soil moisture content. d) The coefficient of kinetiic friction of iron plate on artificial soil was 0.31 to 0.41 and was comparable with that of the natural soil. e) When the pulling speed of soil bin was the 2nd forward speed of power tiller, the rpm of driving shaft of rotary was similar to that of power tiller, soil bin apparatus is indicating the good indoor tester. 3. The comparative experiment on the power requirement of prototype tine of reverse rotating rotary: According to the preliminary test of rotary tine developed with various degrees of angle between holder and edge pcint due to the kinematic analysis, comparative test between prototype rotary tine with $30 ^\circ $ and $10 ^\circ$ of it and presently being used rotary tine was carried out 2nd the results were as follows: a) The total cutting torque was low when the angle between holder and edge point was reduced. b) $\theta r$ (angle between holder and edge point) of rotary tine seemed to be one: of the factors maximizing the increase of torque. c) As the angle between holder and edge point ($\theta r$) of rotary tine was $30 ^\circ $ rather than $45 ^\circ $, the angle of rotation during cutting soil was reduced and the total cutting torque was accordingly reduced about 10%, and the reduction efficiency of total cutting torque was low when the angle between holder and edge point ($\theta r$) of rotary tine was $10 ^\circ $, which indicates that the proper angle between holder and edge point of rotary tine should be larger than $10 ^\circ $ and smaller than $30 ^\circ $ . From above results, it could be concluded that the use of the prototype rotary tine which reduced the angle between holder and edge point to $30 ^\circ $, insted of $45 ^\circ $, is disirable not only decreasing the power requirements, but also increasing the durabie hour of it. Also forward researches are needed, WIlich determine the optimum tilted angle of rotary brocket, and rearrangement of the rotary tine on the rotary boss.

• Microbiology and Biotechnology Letters
• /
• v.26 no.2
• /
• pp.122-129
• /
• 1998

It were reported that antifungal mechanism of Enterobacter cloacae is a volatile ammonia that produced by the strain in soil, and the production of ammonia is related to the bacterial urease activity. A powerful bacterium SH14 against soil-borne pathogen Fusarium solani, which cause root rot of many important crops, was selected from a ginseng pathogen suppressive soil. The strain SH14 was identified as Bacillus subtilis by cultural, biochemical, morphological method, and $API^{circledR}$ test. From several in vitro tests, the antifungal substance that is produced from B. subtilis SH14 was revealed as heat-stable and low-molecular weight antibiotic substance. In order to construct the multifunctional biocontrol agent, the urease gene of Bacillus pasteurii which can produce pathogenes-suppressive ammonia transferred into antifungal bacterium. First, a partial BamH I digestion fragment of plasmid pBU11 containing the alkalophilic B. pasteurii l1859 urease gene was inserted into the BamH I site of pEB203 and expressed in Escherichia coli JM109. The recombinant plasmid was designated as pGU366. The plasmid pGU366 containing urease gene was introduced into the B. subtilis SH14 with PEG-induced protoplast transformation (PIP) method. The urease gene was very stably expressed in the transformant of B. subtilis SH14. Also, the optimal conditions for transformation were established and the highest transformation frequency was obtained by treatment of lysozyme for 90 min, and then addition of 1.5 ${mu}g$/ml DNA and 40% PEG4000. From the in vitro antifungal test against F. solani, antifungal activity of B. subtilis SH14(pGu366) containing urease gene was much higher than that of the host strain. Genetical development of B. subtilis SH14 by transfer of urease gene can be responsible for enhanced biocontrol efficacy with its antibiotic action.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.31 no.3
• /
• pp.199-218
• /
• 2005

Purpose of Study: Peripheral nerve regeneration depends on neurotrophism of distal nerve stump, recovery potential of neuron, supporting cell like Schwann cell and neurotrophic factors such as BDNF. Peripheral nerve regeneration can be enhanced by the conduit which connects the both sides of transected nerve. The conduit maintains the effects of neurotrophism and BDNF produced by Schwann cells which can be made by gene therapy. In this study, we tried to enhance the peripheral nerve regeneration by using calcium phosphate coated porous conduit and BDNF-Adenovirus infected Schwann cells in sciatic nerve of rats. Materials and Methods: Microporous filter which permits the tissue fluid essential for nerve regeneration and does not permit infiltration of fibroblasts, was made into 2mm diameter and 17mm length conduit. Then it was coated with calcium phosphate to improve the Schwann cell adhesion and survival. The coated filter was evaluated by SEM examination and MTT assay. For effective allogenic Schwann cell culture, dorsal root ganglia of 1-day old rat were extracted and treated with enzyme and antimitotic Ara-C. Human BDNF cDNA was obtained from cDNA library and amplified using PCR. BDNF gene was inserted into adenovirus shuttle vector pAACCMVpARS in which E1 was deleted. We infected the BDNF-Ad into 293 human mammary kidney cell-line and obtained the virus plaque 2 days later. RT-PCR was performed to evaluate the secretion of BDNF in infected Schwann cells. To determine the most optimal m.o.i of BDNF-Ad, we infected the Schwann cells with LacZ adenovirus in 1, 20, 50, 75, 100, 250 m.o.i for 2 hours and stained with ${\beta}$-galactosidase. Rats(n=24) weighing around 300g were used. Total 14mm sciatic nerve defect was made and connected with calcium phosphate coated conduits. Schwann cells$(1{\times}10^6)$ or BDNF-Ad infected Schwann cells$(1{\times}10^6)$ were injected in conduit and only media(MEM) was injected in control group. Twelve weeks after surgery, degree of nerve regeneration was evaluated with gait analysis, electrophysiologic measurements and histomorphometric analysis. Results: 1. Microporous Millipore filter was effective conduit which permitted the adhesion of Schwann cells and inhibited the adhesion of fibroblast. We could enhance the Schwann cell adhesion and survival by coating Millipore filter with calcium phosphate. 2. Schwann cell culture technique using repeated treatment of Ara-C and GDNF was established. The mean number of Schwann cells obtained 1 and 2 weeks after the culture were $1.54{\pm}4.0{\times}10^6$ and $9.66{\pm}9.6{\times}10^6$. 3. The mRNA of BDNF in BDNF-Ad infected Schwann cells was detected using RT-PCR. In Schwann cell $0.69\;{\mu}g/{\mu}l$ of DNA was detected and in BDNF-Adenovirus transfected Schwann cell $0.795\;{\mu}g/{\mu}l$ of DNA was detected. The most effective infection concentration was determined by LacZ Adenovirus and 75 m.o.i was found the most optimal. Conclusion: BDNF-Ad transfected Schwann cells successfully regenerated the 14mm nerve gap which was connected with calcium phosphate coated Millipore filter. The BDNF-Ad group showed better results compared with Schwann cells only group and control group in aspect to sciatic function index, electrophysiologic measurements and histomorphometric analysis.